|
1
|
Tong B, Yu Y, Shi S. Rhodotorula sp. as a promising host for microbial cell factories. Metab Eng 2025; 90:178-196. [PMID: 40139654 DOI: 10.1016/j.ymben.2025.03.015] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/15/2024] [Revised: 03/15/2025] [Accepted: 03/21/2025] [Indexed: 03/29/2025]
Abstract
Rhodotorula sp. is a red yeast that has emerged as a promising host for microbial cell factories. Under specific conditions, Rhodotorula sp. can accumulate lipids that constitute over 70% of its dry cell weight, underscoring its potential in lipid compound production. Additionally, it can utilize a variety of carbon sources, including glucose, xylose, and volatile fatty acids, and exhibits high tolerance to low-cost carbon sources and industrial by-products, showcasing its excellent performance in industrial processes. Furthermore, the native mevalonate pathway of Rhodotorula sp. enables its efficient synthesis of antioxidant carotenoids and other terpenoids, which are widely applied in the food, pharmaceutical, and cosmetic industries. Due to its excellent accumulation ability of lipophilic compounds, metabolic diversity, and environmental adaptability, this review summarizes recent advances in genetic elements and metabolic engineering technologies for Rhodotorula sp., emphasizing its potential as a chassis cell factory for the production of lipids, carotenoids, and other chemicals. It also highlights key factors influencing commercial fermentation processes and concludes with challenges and solutions for further developing Rhodotorula sp. as microbial chassis.
Collapse
Affiliation(s)
- Baisong Tong
- Beijing Advanced Innovation Center for Soft Matter Science and Engineering, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, 100029, Beijing, China
| | - Yi Yu
- Beijing Advanced Innovation Center for Soft Matter Science and Engineering, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, 100029, Beijing, China
| | - Shuobo Shi
- Beijing Advanced Innovation Center for Soft Matter Science and Engineering, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, 100029, Beijing, China.
| |
Collapse
|
|
2
|
Li YZ, Wang Q, Cheng C, Chen C, Zhang FL, Zou Y, Li J, Zhao XQ. Genome mining of an endophytic natural yeast isolate Rhodotorula sp. Y090 and production of the potent antioxidant ergothioneine. J Biotechnol 2025; 404:18-26. [PMID: 40169100 DOI: 10.1016/j.jbiotec.2025.03.019] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/04/2024] [Revised: 02/19/2025] [Accepted: 03/26/2025] [Indexed: 04/03/2025]
Abstract
Endophytic yeast strains are promising sources of various valuable bioactive compounds. However, studies on endophytic strains derived from lichen samples remain limited. In this study, we isolated and characterized Rhodotorula sp. Y090 from a Usnea longissima lichen sample, and investigated its biosynthetic potential. Genome mining revealed distinct genetic features that differed from its closest relative strain R. graminis WP1. Ergothioneine (EGT) is a potent antioxidant and rare sulfur-containing histidine derivative. However, so far, the EGT biosynthetic enzymes by natural yeast strains have been limitedly studied. In this study, combining genome mining and transcriptomic analysis, genes encoding the potential enzymes for the production of EGT and xylose utilization were identified in Rhodotorula sp. Y090. Further studies demonstrated that Rhodotorula sp. Y090 was capable of producing EGT using xylose, glucose, glycerol, or sucrose as the sole carbon source, and the highest titer reached 363.6 mg/L in shake flask culture, which is significantly higher than that of the most reported levels in the other natural yeasts. Rhodotorula sp. Y090 also exhibited a good ability of EGT export, which could facilitate cost-effective production. These findings suggest that the lichen-derived endophytic yeast Rhodotorula sp. Y090 represents a promising natural candidate for bio-production of the potent antioxidant.
Collapse
Affiliation(s)
- Yu-Zhen Li
- State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China
| | - Qi Wang
- State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China
| | - Cheng Cheng
- School of Biotechnology and Food Engineering, Hefei Normal University, Hefei 230601, China
| | - Chao Chen
- College of Life Science, Dalian Minzu University, Dalian, Liaoning 116600, China
| | - Feng-Li Zhang
- State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China
| | - Yue Zou
- Shanghai CHANDO Group Co., Ltd, Shanghai 200233, China
| | - Jun Li
- Shanghai CHANDO Group Co., Ltd, Shanghai 200233, China.
| | - Xin-Qing Zhao
- State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.
| |
Collapse
|
|
3
|
Fonseca RSK, Lotas KM, Cortez ACA, Fernandes FDS, de Souza ÉS, Dufossé L, de Souza JVB. Exploration of carotenoid-producing Rhodotorula yeasts from amazonian substrates for sustainable biotechnology applications. CURRENT RESEARCH IN MICROBIAL SCIENCES 2025; 8:100373. [PMID: 40177628 PMCID: PMC11964568 DOI: 10.1016/j.crmicr.2025.100373] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 04/05/2025] Open
Abstract
Carotenoids are natural pigments responsible for the yellow, orange, and red colors seen in various organisms. The aim of this study was to investigate the potential of yeast strains belonging to the genus Rhodotorula isolated from the Amazon Region in the production of carotenoids. Environmental samples from the Amazon Region were collected for yeast isolation. Isolates showing pigmented colonies underwent morphological and biochemical studies, as well as assessment of their potential for carotenoid production. The three best producers were identified through nucleotide sequencing of the ITS1-5.8s-ITS4 rDNA region. The top producer underwent univariate experiments to evaluate the influence of different C/N sources. Carotenoids produced were evaluated using CCD. Results showed the isolation of 13 pigmented yeasts with morphological and biochemical characteristics consistent with the genus Rhodotorula. Isolates RGM42 (601 μg/g), RTC42 (362 μg/g), and RTC45 (351 μg/g) stood out as the top carotenoid producers. These yeasts were identified as belonging to the species Rhodotorula mucilaginosa. Regarding C/N influence, R. mucilaginosa RGM42 produced the highest number of carotenoids using glucose and peptone as carbon and nitrogen sources, respectively. Isolate R. mucilaginosa RGM42 exhibited maximum growth with a peak at 72 h of bioprocess. Extracts from isolates R. mucilaginosa RGM42, R. mucilaginosa RTC42, and R. mucilaginosa RTC45 showed retention indices like β-carotene in thin-layer chromatography; however, R. mucilaginosa RGM42 presented an additional pigment in its chromatographic profile.
Collapse
Affiliation(s)
- Raissa Sayumy Kataki Fonseca
- Programa de Pós-Graduação em Biotecnologia e Recursos Naturais (PPG-MBT) na Universidade do Estado do Amazonas (UEA), Manaus, Amazonas, Brazil
| | - Kevyn Melo Lotas
- Programa de Pós-Graduação em Biotecnologia e Recursos Naturais (PPG-MBT) na Universidade do Estado do Amazonas (UEA), Manaus, Amazonas, Brazil
| | - Ana Claudia Alves Cortez
- Instituto Nacional de Pesquisas da Amazônia – INPA. Av. André Araújo, 2936 - Petrópolis, 69067-375, Manaus AM, Brazil
| | - Flávia da Silva Fernandes
- Instituto Nacional de Pesquisas da Amazônia – INPA. Av. André Araújo, 2936 - Petrópolis, 69067-375, Manaus AM, Brazil
| | - Érica Simplício de Souza
- Universidade do Estado do Amazonas – UEA. Av. Carvalho Leal, 1777 - Cachoeirinha, 69065-001, Manaus AM, Brazil
| | - Laurent Dufossé
- CHEMBIOPRO Lab, Chimie et Biotechnologie des Produits Naturels, ESIROI Agroalimentaire, University of Réunion Island, 97400 Saint-Denis, France
| | - João Vicente Braga de Souza
- Instituto Nacional de Pesquisas da Amazônia – INPA. Av. André Araújo, 2936 - Petrópolis, 69067-375, Manaus AM, Brazil
| |
Collapse
|
|
4
|
Gong G, Wu B, Liu L, Li J, He M. Engineering oleaginous red yeasts as versatile chassis for the production of oleochemicals and valuable compounds: Current advances and perspectives. Biotechnol Adv 2024; 76:108432. [PMID: 39163921 DOI: 10.1016/j.biotechadv.2024.108432] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/11/2024] [Revised: 07/04/2024] [Accepted: 08/16/2024] [Indexed: 08/22/2024]
Abstract
Enabling the transition towards a future circular bioeconomy based on industrial biomanufacturing necessitates the development of efficient and versatile microbial platforms for sustainable chemical and fuel production. Recently, there has been growing interest in engineering non-model microbes as superior biomanufacturing platforms due to their broad substrate range and high resistance to stress conditions. Among these non-conventional microbes, red yeasts belonging to the genus Rhodotorula have emerged as promising industrial chassis for the production of specialty chemicals such as oleochemicals, organic acids, fatty acid derivatives, terpenoids, and other valuable compounds. Advancements in genetic and metabolic engineering techniques, coupled with systems biology analysis, have significantly enhanced the production capacity of red yeasts. These developments have also expanded the range of substrates and products that can be utilized or synthesized by these yeast species. This review comprehensively examines the current efforts and recent progress made in red yeast research. It encompasses the exploration of available substrates, systems analysis using multi-omics data, establishment of genome-scale models, development of efficient molecular tools, identification of genetic elements, and engineering approaches for the production of various industrially relevant bioproducts. Furthermore, strategies to improve substrate conversion and product formation both with systematic and synthetic biology approaches are discussed, along with future directions and perspectives in improving red yeasts as more versatile biotechnological chassis in contributing to a circular bioeconomy. The review aims to provide insights and directions for further research in this rapidly evolving field. Ultimately, harnessing the capabilities of red yeasts will play a crucial role in paving the way towards next-generation sustainable bioeconomy.
Collapse
Affiliation(s)
- Guiping Gong
- Biomass Energy Technology Research Centre, Rural Energy and Ecology Research Center of CAAS, Key Laboratory of Development and Application of Rural Renewable Energy (Ministry of Agriculture and Rural Affairs), Biogas Institute of Ministry of Agriculture and Rural Affairs, Chengdu 610041, PR China.
| | - Bo Wu
- Biomass Energy Technology Research Centre, Rural Energy and Ecology Research Center of CAAS, Key Laboratory of Development and Application of Rural Renewable Energy (Ministry of Agriculture and Rural Affairs), Biogas Institute of Ministry of Agriculture and Rural Affairs, Chengdu 610041, PR China
| | - Linpei Liu
- Biomass Energy Technology Research Centre, Rural Energy and Ecology Research Center of CAAS, Key Laboratory of Development and Application of Rural Renewable Energy (Ministry of Agriculture and Rural Affairs), Biogas Institute of Ministry of Agriculture and Rural Affairs, Chengdu 610041, PR China
| | - Jianting Li
- Biomass Energy Technology Research Centre, Rural Energy and Ecology Research Center of CAAS, Key Laboratory of Development and Application of Rural Renewable Energy (Ministry of Agriculture and Rural Affairs), Biogas Institute of Ministry of Agriculture and Rural Affairs, Chengdu 610041, PR China
| | - Mingxiong He
- Biomass Energy Technology Research Centre, Rural Energy and Ecology Research Center of CAAS, Key Laboratory of Development and Application of Rural Renewable Energy (Ministry of Agriculture and Rural Affairs), Biogas Institute of Ministry of Agriculture and Rural Affairs, Chengdu 610041, PR China
| |
Collapse
|
|
5
|
Mummaleti G, Udo T, Mohan A, Kong F. Synthesis, characterization and application of microbial pigments in foods as natural colors. Crit Rev Food Sci Nutr 2024:1-30. [PMID: 39466660 DOI: 10.1080/10408398.2024.2417802] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/30/2024]
Abstract
Colorants have played a crucial role in various applications, particularly in food processing, with natural sources such as mineral ores, plants, insects, and animals being commonly used. However, the nineteenth century saw the development of synthetic dyes, which replaced these natural colorants. In recent years, there has been a growing demand for natural products, driving an increased interest in natural colorants. Microbial pigments have emerged as promising sources of natural pigments due to their numerous health benefits. They can be produced in large quantities rapidly and from more affordable substrates, making them economically attractive. This review focuses on the current advancements in the low-cost synthesis of microbial pigments, exploring their biological activities and commercial applications. Microbial pigments offer a sustainable and economically viable alternative to natural and synthetic colorants, meeting the growing demand for natural products. These pigments are relatively nontoxic and exhibit significant health benefits, making them suitable for a wide range of applications. As interest in natural products continues to rise, microbial pigments hold great potential in shaping the future of colorant production across various sectors.
Collapse
Affiliation(s)
- Gopinath Mummaleti
- Department of Food Science and Technology, The University of Georgia, Athens, Georgia, USA
| | - Toshifumi Udo
- Department of Food Science and Technology, The University of Georgia, Athens, Georgia, USA
| | - Anand Mohan
- Department of Food Science and Technology, The University of Georgia, Athens, Georgia, USA
| | - Fanbin Kong
- Department of Food Science and Technology, The University of Georgia, Athens, Georgia, USA
| |
Collapse
|
|
6
|
He Q, Bai S, Chen C, Yang X, Li Z, Sun S, Qu X, Yang X, Pan J, Liu W, Hou C, Deng Y. A chromosome-scale genome provides new insights into the typical carotenoid biosynthesis in the important red yeast Rhodotorula glutinis QYH-2023 with anti-inflammatory effects. Int J Biol Macromol 2024; 269:132103. [PMID: 38719011 DOI: 10.1016/j.ijbiomac.2024.132103] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/24/2024] [Revised: 04/30/2024] [Accepted: 05/03/2024] [Indexed: 05/12/2024]
Abstract
Rhodotorula spp. has been studied as one powerful source for a novel cell factory with fast growth and its high added-value biomolecules. However, its inadequate genome and genomic annotation have hindered its widespread use in cosmetics and food industries. Rhodotorula glutinis QYH-2023, was isolated from rice rhizosphere soil, and the highest quality of the genome of the strain was obtained at chromosome level (18 chromosomes) than ever before in red yeast in this study. Comparative genomics analysis revealed that there are more key gene copies of carotenoids biosynthesis in R. glutinis QYH-2023 than other species of Rhodotorula spp. Integrated transcriptome and metabolome analysis revealed that lipids and carotenoids biosynthesis was significantly enriched during fermentation. Subsequent investigation revealed that the over-expression of the strain three genes related to carotenoids biosynthesis in Komagataella phaffii significantly promoted the carotenoid production. Furthermore, in vitro tests initially confirmed that the longer the fermentation period, the synthesized metabolites controlled by R. glutinis QYH-2023 genome had the stronger anti-inflammatory properties. All of the findings revealed a high-quality reference genome which highlight the potential of R. glutinis strains to be employed as chassis cells for biosynthesizing carotenoids and other active chemicals.
Collapse
Affiliation(s)
- Qiaoyun He
- Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha 410205, PR China; Graduate School of Chinese Academy of Agricultural Sciences, Beijing 100081, PR China
| | - Shasha Bai
- Beijing Institute of Nutritional Resources Co., LTD, Beijing Academy of Science and Technology, Beijing 100069, PR China; Department of Biomedical Sciences, Beijing city university, Beijing 100083, PR China
| | - Chenxiao Chen
- Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha 410205, PR China
| | - Xiai Yang
- Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha 410205, PR China
| | - Zhimin Li
- Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha 410205, PR China
| | - Shitao Sun
- Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha 410205, PR China
| | - Xiaoxin Qu
- Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha 410205, PR China
| | - Xiushi Yang
- Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha 410205, PR China
| | - Jiangpeng Pan
- Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha 410205, PR China
| | - Wei Liu
- Hangzhou Base Array Biotechnology Co., Ltd., Hangzhou 310000, PR China
| | - Chunsheng Hou
- Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha 410205, PR China.
| | - Yanchun Deng
- Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha 410205, PR China.
| |
Collapse
|
|
7
|
Gallego-García M, Susmozas A, Negro MJ, Moreno AD. Challenges and prospects of yeast-based microbial oil production within a biorefinery concept. Microb Cell Fact 2023; 22:246. [PMID: 38053171 DOI: 10.1186/s12934-023-02254-4] [Citation(s) in RCA: 2] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/06/2023] [Accepted: 11/17/2023] [Indexed: 12/07/2023] Open
Abstract
Biodiesel, unlike to its fossil-based homologue (diesel), is renewable. Its use contributes to greater sustainability in the energy sector, mainly by reducing greenhouse gas emissions. Current biodiesel production relies on plant- and animal-related feedstocks, resulting in high final costs to the prices of those raw materials. In addition, the production of those materials competes for arable land and has provoked a heated debate involving their use food vs. fuel. As an alternative, single-cell oils (SCOs) obtained from oleaginous microorganisms are attractive sources as a biofuel precursor due to their high lipid content, and composition similar to vegetable oils and animal fats. To make SCOs competitive from an economic point of view, the use of readily available low-cost substrates becomes essential. This work reviews the most recent advances in microbial oil production from non-synthetic sugar-rich media, particularly sugars from lignocellulosic wastes, highlighting the main challenges and prospects for deploying this technology fully in the framework of a Biorefinery concept.
Collapse
Affiliation(s)
- María Gallego-García
- Advanced Biofuels and Bioproducts Unit, Department of Energy, Research Center for Energy, Environment and Technology (CIEMAT), Avda. Complutense 40, Madrid, 28040, Spain
- Department of Biomedicine and Biotechnology, University of Alcalá de Henares, Alcalá de Henares, Spain
| | - Ana Susmozas
- Advanced Biofuels and Bioproducts Unit, Department of Energy, Research Center for Energy, Environment and Technology (CIEMAT), Avda. Complutense 40, Madrid, 28040, Spain
| | - María José Negro
- Advanced Biofuels and Bioproducts Unit, Department of Energy, Research Center for Energy, Environment and Technology (CIEMAT), Avda. Complutense 40, Madrid, 28040, Spain.
| | - Antonio D Moreno
- Advanced Biofuels and Bioproducts Unit, Department of Energy, Research Center for Energy, Environment and Technology (CIEMAT), Avda. Complutense 40, Madrid, 28040, Spain
| |
Collapse
|
|
8
|
Rocha Balbino T, Sánchez-Muñoz S, Díaz-Ruíz E, Moura Rocha T, Mier-Alba E, Custódio Inácio S, Jose Castro-Alonso M, de Carvalho Santos-Ebinuma V, Fernando Brandão Pereira J, César Santos J, Silvério da Silva S. Lignocellulosic biorefineries as a platform for the production of high-value yeast derived pigments - A review. BIORESOURCE TECHNOLOGY 2023; 386:129549. [PMID: 37499926 DOI: 10.1016/j.biortech.2023.129549] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 05/30/2023] [Revised: 07/21/2023] [Accepted: 07/22/2023] [Indexed: 07/29/2023]
Abstract
Lignocellulosic byproducts, mainly generated by the agro-industrial sector, have great potential as cost-effective feedstocks for bioprocesses because of their abundant availability and high content of sugar-rich and nutrient-rich elements. This biomass can be employed as a carbon source to produce various molecules using several microorganisms. Yeast strains have shown their capability to metabolize diverse C5 and C6 carbon sources, thereby facilitating their use in the bioprocessing of lignocellulosic biomass. Furthermore, yeasts can produce a wide range of valuable products, including biofuels, enzymes, proteins, and pigments, making them attractive for use in integrated biorefineries. Yeast-derived pigments have versatile applications and are environmentally friendly alternatives to their synthetic counterparts. This review emphasizes the potential of lignocellulosic biomass as a feedstock for producing yeast-derived products with a focus on pigments as valuable molecules. It also proposes a yeast-derived pigment platform utilizing lignocellulosic byproducts and explores its potential integration in biorefineries.
Collapse
Affiliation(s)
- Thercia Rocha Balbino
- Bioprocesses and Sustainable Products Laboratory, Department of Biotechnology, Engineering School of Lorena, University of São Paulo (EEL-USP), 12.602.810. Lorena, São Paulo, Brazil.
| | - Salvador Sánchez-Muñoz
- Bioprocesses and Sustainable Products Laboratory, Department of Biotechnology, Engineering School of Lorena, University of São Paulo (EEL-USP), 12.602.810. Lorena, São Paulo, Brazil
| | - Erick Díaz-Ruíz
- Bioprocesses and Sustainable Products Laboratory, Department of Biotechnology, Engineering School of Lorena, University of São Paulo (EEL-USP), 12.602.810. Lorena, São Paulo, Brazil
| | - Thiago Moura Rocha
- Bioprocesses and Sustainable Products Laboratory, Department of Biotechnology, Engineering School of Lorena, University of São Paulo (EEL-USP), 12.602.810. Lorena, São Paulo, Brazil
| | - Edith Mier-Alba
- Bioprocesses and Sustainable Products Laboratory, Department of Biotechnology, Engineering School of Lorena, University of São Paulo (EEL-USP), 12.602.810. Lorena, São Paulo, Brazil
| | - Stephanie Custódio Inácio
- Bioprocesses and Sustainable Products Laboratory, Department of Biotechnology, Engineering School of Lorena, University of São Paulo (EEL-USP), 12.602.810. Lorena, São Paulo, Brazil
| | - Maria Jose Castro-Alonso
- Bioprocesses and Sustainable Products Laboratory, Department of Biotechnology, Engineering School of Lorena, University of São Paulo (EEL-USP), 12.602.810. Lorena, São Paulo, Brazil
| | - Valéria de Carvalho Santos-Ebinuma
- School of Pharmaceutical Sciences, Department of Bioprocess Engineering and Biotechnology, São Paulo State University (UNESP), Araraquara, São Paulo 14801-902, Brazil
| | - Jorge Fernando Brandão Pereira
- University of Coimbra, CIEPQPF, FCTUC, Department of Chemical Engineering, Rua Sílvio Lima, Pólo II - Pinhal de Marrocos, 3030-790, Coimbra 30-790, Portugal
| | - Júlio César Santos
- Laboratory of Biopolymers, Bioreactors and Process Simulation, Department of Biotechnology, Engineering School of Lorena, University of São Paulo (EEL-USP), 12.602.810. Lorena, São Paulo, Brazil
| | - Silvio Silvério da Silva
- Bioprocesses and Sustainable Products Laboratory, Department of Biotechnology, Engineering School of Lorena, University of São Paulo (EEL-USP), 12.602.810. Lorena, São Paulo, Brazil
| |
Collapse
|
|
9
|
Mussagy CU, Ribeiro HF, Pereira JFB. Rhodotorula sp. as a cell factory for production of valuable biomolecules. ADVANCES IN APPLIED MICROBIOLOGY 2023; 123:133-156. [PMID: 37400173 DOI: 10.1016/bs.aambs.2023.04.001] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 07/05/2023]
Abstract
Rhodotorula sp. are well-known for their ability to biosynthesize a diverse range of valuable biomolecules, including carotenoids, lipids, enzymes, and polysaccharides. Despite the high number of studies conducted using Rhodotorula sp. at the laboratory scale, most of these do not address all processual aspects necessary for scaling up these processes for industrial applications. This chapter explores the potential of Rhodotorula sp. as a cell factory for the production of distinct biomolecules, with a particular emphasis on exploring their use from a biorefinery perspective. Through in-depth discussions of the latest research and insights into non-conventional applications, we aim to provide a comprehensive understanding of Rhodotorula sp.'s ability to produce biofuels, bioplastics, pharmaceuticals, and other valuable biochemicals. This book chapter also examines the fundamentals and challenges associated with the optimizing upstream and downstream processing of Rhodotorula sp-based processes. We believe that through this chapter, readers with different levels of expertise will gain insights into strategies for enhancing the sustainability, efficiency, and effectiveness of producing biomolecules using Rhodotorula sp.
Collapse
Affiliation(s)
- Cassamo U Mussagy
- Escuela de Agronomía, Facultad de Ciencias Agronómicas y de los Alimentos, Pontificia Universidad Católica de Valparaíso, Quillota, Chile.
| | - Helena F Ribeiro
- Department of Chemical Engineering, University of Coimbra, CIEPQPF, Coimbra, Portugal
| | - Jorge F B Pereira
- Department of Chemical Engineering, University of Coimbra, CIEPQPF, Coimbra, Portugal
| |
Collapse
|
|
10
|
Li J, Chen Y, Gao A, Wei L, Wei D, Wang W. Simultaneous Production of Cellulase and β-Carotene in the Filamentous Fungus Trichoderma reesei. JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 2023; 71:6358-6365. [PMID: 37042195 DOI: 10.1021/acs.jafc.3c00690] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 05/03/2023]
Abstract
β-Carotene is an indispensable additive in beverage, cosmetic, feed, and pharmaceutical production. The fermentation industry annually generates abundant waste mycelia from Trichoderma reesei (T. reesei), a pivotal industrial strain for cellulase and heterologous protein production. In this study, we constructed a T. reesei cell factory for β-carotene production for the first time. Four key enzymes, CarRP, CarB, GGS1/CrtE, and HMG1, were overexpressed in T. reesei. The concentrations of medium components, including tryptone and glucose, were optimized. The modified strain accumulated β-carotene at a titer of 218.8 mg/L in flask culture. We achieved cellulase production (FPase, 22.33 IU/mL) with the concomitant production of β-carotene (286.63 mg/L) from T. reesei in a jar. Overall, this study offers a novel and unique approach to address the costly waste mycelium management process using T. reesei industrial strains that simultaneously produce proteins and carotenoids.
Collapse
Affiliation(s)
- Jing Li
- State Key Lab of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China
| | - Yumeng Chen
- State Key Lab of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China
| | - Ao Gao
- State Key Lab of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China
| | - Liujing Wei
- State Key Lab of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China
| | - Dongzhi Wei
- State Key Lab of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China
| | - Wei Wang
- State Key Lab of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China
| |
Collapse
|
|
11
|
Watcharawipas A, Runguphan W. Red yeasts and their carotenogenic enzymes for microbial carotenoid production. FEMS Yeast Res 2023; 23:6895548. [PMID: 36513367 DOI: 10.1093/femsyr/foac063] [Citation(s) in RCA: 6] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/24/2022] [Revised: 12/08/2022] [Accepted: 12/12/2022] [Indexed: 12/15/2022] Open
Abstract
Carotenoids are C40 isoprene-based compounds with significant commercial interests that harbor diverse bioactivities. Prominent examples of carotenoids are beta-carotene, a precursor to vitamin A essential for proper eye health, and lycopene and astaxanthin, powerful antioxidants implicated in preventing cancers and atherosclerosis. Due to their benefits to human health, the market value for carotenoids is rapidly increasing and is projected to reach USD 1.7 billion by 2025. However, their production now relies on chemical synthesis and extraction from plants that pose risks to food management and numerous biological safety issues. Thus, carotenoid production from microbes is considered a promising strategy for achieving a healthy society with more sustainability. Red yeast is a heterogeneous group of basidiomycetous fungi capable of producing carotenoids. It is a critical source of microbial carotenoids from low-cost substrates. Carotenogenic enzymes from red yeasts have also been highly efficient, invaluable biological resources for biotechnological applications. In this minireview, we focus on red yeast as a promising source for microbial carotenoids, strain engineering strategies for improving carotenoid production in red yeasts, and potential applications of carotenogenic enzymes from red yeasts in conventional and nonconventional yeasts.
Collapse
Affiliation(s)
- Akaraphol Watcharawipas
- Department of Microbiology, Faculty of Science, Mahidol University, 272 Rama VI Road, Ratchathewi, Bangkok 10400, Thailand
| | - Weerawat Runguphan
- National Center for Genetic Engineering and Biotechnology, 113 Thailand Science Park, Paholyothin Road, Klong 1, Klong Luang, Pathumthani 12120, Thailand
| |
Collapse
|
|
12
|
Li Z, Li C, Cheng P, Yu G. Rhodotorula mucilaginosa-alternative sources of natural carotenoids, lipids, and enzymes for industrial use. Heliyon 2022. [PMID: 36419653 DOI: 10.1016/j.heliyon.2022.e1150] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 05/05/2023] Open
Abstract
Biotechnologically useful yeast strains have been receiving important attention worldwide for the demand of a wide range of industries. Rhodotorula mucilaginosa is recognized as a biotechnologically important yeast that has gained great interest as a promising platform strain, owing to the diverse substrate appetites, robust stress resistance, and other gratifying features. Due to its attractive properties, R. mucilaginosa has been regarded as an excellent candidate for the biorefinery of carotenoids, lipids, enzymes, and other functional bioproducts by utilizing low-cost agricultural waste materials as substrates. These compounds have aroused great interest as the potential alternative sources of health-promoting food products, substrates for so-called third-generation biodiesel, and dyes or functional ingredients for cosmetics. Furthermore, the use of R. mucilaginosa has rapidly increased as a result of advancements in fermentation for enhanced production of these valuable bioactive compounds. This review focuses on R. mucilaginosa in these advancements and summarizes its potential prospects as alternative sources of natural bioproducts.
Collapse
Affiliation(s)
- Zhiheng Li
- Innovative Institute for Plant Health, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China
- College of Agriculture and Biology, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China
| | - Chunji Li
- Innovative Institute for Plant Health, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China
- College of Agriculture and Biology, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China
| | - Ping Cheng
- Innovative Institute for Plant Health, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China
- College of Agriculture and Biology, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China
| | - Guohui Yu
- Innovative Institute for Plant Health, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China
- College of Agriculture and Biology, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China
| |
Collapse
|
|
13
|
Li Z, Li C, Cheng P, Yu G. Rhodotorula mucilaginosa—alternative sources of natural carotenoids, lipids, and enzymes for industrial use. Heliyon 2022; 8:e11505. [DOI: 10.1016/j.heliyon.2022.e11505] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/26/2022] [Revised: 08/19/2022] [Accepted: 11/04/2022] [Indexed: 11/16/2022] Open
|
|
14
|
Deeba F, Kiran Kumar K, Ali Wani S, Kumar Singh A, Sharma J, Gaur NA. Enhanced biodiesel and β-carotene production in Rhodotorula pacifica INDKK using sugarcane bagasse and molasses by an integrated biorefinery framework. BIORESOURCE TECHNOLOGY 2022; 351:127067. [PMID: 35351564 DOI: 10.1016/j.biortech.2022.127067] [Citation(s) in RCA: 18] [Impact Index Per Article: 6.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 01/31/2022] [Revised: 03/22/2022] [Accepted: 03/23/2022] [Indexed: 06/14/2023]
Abstract
Dependency on fossil fuels raises an economic and ecological concern that has urged to look for alternative sources of energy. Bio-refinery concept is one of the alternate frameworks for the biomass conversion into biofuel and other value-added by-products. The present work illustrates importance of an oleaginous yeast Rhodotorula pacifica INDKK in an integrated bio-refinery field by utilizing renewable sugars generated from lignocellulosic biomass. The maximum 11.8 g/L lipid titer, 210.4 mg/L β-carotene and 7.1 g animal feed were produced by R. pacifica INDKK in bioreactor containing 5% (v/v) molasses supplemented with enzymatically hydrolyzed and alkali-pretreated sugarcane bagasse hydrolysate (35% v/v). Furthermore, xylooligosaccharides (20.6 g/L), a beneficial prebiotics were also produced from the hemicellulosic fraction separated after alkali pretreatment of bagasse. This novel concept of integrated yeast bio-refinery for concomitant production of biodiesel and multiple value-added products with minimum waste generation is proposed as a sustainable and profitable process.
Collapse
Affiliation(s)
- Farha Deeba
- Yeast Biofuel Group, DBT-ICGEB Centre for Advanced Bioenergy Research, International Centre for Genetic Engineering and Biotechnology, New Delhi 110067, India
| | - Kukkala Kiran Kumar
- Yeast Biofuel Group, DBT-ICGEB Centre for Advanced Bioenergy Research, International Centre for Genetic Engineering and Biotechnology, New Delhi 110067, India
| | - Shahid Ali Wani
- Yeast Biofuel Group, DBT-ICGEB Centre for Advanced Bioenergy Research, International Centre for Genetic Engineering and Biotechnology, New Delhi 110067, India
| | - Anup Kumar Singh
- Yeast Biofuel Group, DBT-ICGEB Centre for Advanced Bioenergy Research, International Centre for Genetic Engineering and Biotechnology, New Delhi 110067, India
| | - Juhi Sharma
- Yeast Biofuel Group, DBT-ICGEB Centre for Advanced Bioenergy Research, International Centre for Genetic Engineering and Biotechnology, New Delhi 110067, India
| | - Naseem A Gaur
- Yeast Biofuel Group, DBT-ICGEB Centre for Advanced Bioenergy Research, International Centre for Genetic Engineering and Biotechnology, New Delhi 110067, India.
| |
Collapse
|
|
15
|
Industrial-Scale Production of Mycotoxin Binder from the Red Yeast Sporidiobolus pararoseus KM281507. J Fungi (Basel) 2022; 8:jof8040353. [PMID: 35448584 PMCID: PMC9029514 DOI: 10.3390/jof8040353] [Citation(s) in RCA: 7] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/07/2022] [Revised: 03/24/2022] [Accepted: 03/28/2022] [Indexed: 02/06/2023] Open
Abstract
Red yeast Sporidiobolus pararoseus KM281507 has been recognized as a potential feed additive. Beyond their nutritional value (carotenoids and lipids), red yeast cells (RYCs) containing high levels of β-glucan can bind mycotoxins. This study investigated the industrial feasibility of the large-scale production of RYCs, along with their ability to act as a mycotoxin binder. Under a semi-controlled pH condition in a 300 L bioreactor, 28.70-g/L biomass, 8.67-g/L lipids, and 96.10-mg/L total carotenoids were obtained, and the RYCs were found to contain 5.73% (w/w) β-glucan. The encapsulated RYC was in vitro tested for its mycotoxin adsorption capacity, including for aflatoxin B1 (AFB1), zearalenone (ZEA), ochratoxin A (OTA), T-2 toxin (T-2) and deoxynivalenol (DON). The RYCs had the highest binding capacity for OTA and T-2 at concentrations of 0.31–1.25 and 0.31–2.5 µg/mL, respectively. The mycotoxin adsorption capacity was further tested using a gastrointestinal poultry model. The adsorption capacities of the RYCs and a commercial mycotoxin binder (CMB) were comparable. The RYCs not only are rich in lipids and carotenoids but also play an important role in mycotoxin binding. Since the industrial-scale production and downstream processing of RYCs were successfully demonstrated, RYCs could be applied as possible feed additives.
Collapse
|
|
16
|
Near Chromosome-Level Genome Assembly and Annotation of Rhodotorula babjevae Strains Reveals High Intraspecific Divergence. J Fungi (Basel) 2022; 8:jof8040323. [PMID: 35448555 PMCID: PMC9027234 DOI: 10.3390/jof8040323] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/26/2021] [Revised: 03/16/2022] [Accepted: 03/19/2022] [Indexed: 02/04/2023] Open
Abstract
The genus Rhodotorula includes basidiomycetous oleaginous yeast species. Rhodotorula babjevae can produce compounds of biotechnological interest such as lipids, carotenoids, and biosurfactants from low value substrates such as lignocellulose hydrolysate. High-quality genome assemblies are needed to develop genetic tools and to understand fungal evolution and genetics. Here, we combined short- and long-read sequencing to resolve the genomes of two R. babjevae strains, CBS 7808 (type strain) and DBVPG 8058, at chromosomal level. Both genomes are 21 Mbp in size and have a GC content of 68.2%. Allele frequency analysis indicates that both strains are tetraploid. The genomes consist of a maximum of 21 chromosomes with a size of 0.4 to 2.4 Mbp. In both assemblies, the mitochondrial genome was recovered in a single contig, that shared 97% pairwise identity. Pairwise identity between most chromosomes ranges from 82 to 87%. We also found indications for strain-specific extrachromosomal endogenous DNA. A total of 7591 and 7481 protein-coding genes were annotated in CBS 7808 and DBVPG 8058, respectively. CBS 7808 accumulated a higher number of tandem duplications than DBVPG 8058. We identified large translocation events between putative chromosomes. Genome divergence values between the two strains indicate that they may belong to different species.
Collapse
|
|
17
|
Carotenoids and Their Biosynthesis in Fungi. Molecules 2022; 27:molecules27041431. [PMID: 35209220 PMCID: PMC8879039 DOI: 10.3390/molecules27041431] [Citation(s) in RCA: 27] [Impact Index Per Article: 9.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/26/2022] [Revised: 02/16/2022] [Accepted: 02/16/2022] [Indexed: 12/04/2022] Open
Abstract
Carotenoids represent a class of pigmented terpenoids. They are distributed in all taxonomic groups of fungi. Most of the fungal carotenoids differ in their chemical structures to those from other organisms. The general function of carotenoids in heterotrophic organisms is protection as antioxidants against reactive oxygen species generated by photosensitized reactions. Furthermore, carotenoids are metabolized to apocarotenoids by oxidative cleavage. This review presents the current knowledge on fungal-specific carotenoids, their occurrence in different taxonomic groups, and their biosynthesis and conversion into trisporic acids. The outline of the different pathways was focused on the reactions and genes involved in not only the known pathways, but also suggested the possible mechanisms of reactions, which may occur in several non-characterized pathways in different fungi. Finally, efforts and strategies for genetic engineering to enhance or establish pathways for the production of various carotenoids in carotenogenic or non-carotenogenic yeasts were highlighted, addressing the most-advanced producers of each engineered yeast, which offered the highest biotechnological potentials as production systems.
Collapse
|
|
18
|
Watcharawipas A, Sansatchanon K, Phithakrotchanakoon C, Tanapongpipat S, Runguphan W, Kocharin K. Novel carotenogenic gene combinations from red yeasts enhanced lycopene and beta-carotene production in Saccharomyces cerevisiae from the low-cost substrate sucrose. FEMS Yeast Res 2021; 21:6449371. [PMID: 34865010 DOI: 10.1093/femsyr/foab062] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/16/2021] [Accepted: 12/01/2021] [Indexed: 12/14/2022] Open
Abstract
Carotenoids (C40H56) including lycopene and beta-carotene are relatively strong antioxidants that provide benefits to human health. Here, we screened highly efficient crt variants from red yeasts to improve lycopene and beta-carotene production in Saccharomyces cerevisiae. We identified that crt variants from Sporidiobolus pararoseus TBRC-BCC 63403 isolated from rice leaf in Thailand exhibited the highest activity in term of lycopene and beta-carotene production in the context of yeast. Specifically, the phytoene desaturase SpCrtI possessed up to 4-fold higher in vivo activity based on lycopene content than the benchmark enzyme BtCrtI from Blakeslea trispora in our engineered WWY005 strain. Also, the geranylgeranyl pyrophosphate (GGPP) synthase SpCrtE, the bifunctional phytoene synthase-lycopene cyclase SpCrtYB, and SpCrtI when combined led to 7-fold improvement in beta-carotene content over the benchmark enzymes from Xanthophyllomyces dendrorhous in the laboratory strain CEN.PK2-1C. Sucrose as an alternative to glucose was found to enhance lycopene production in cells lacking GAL80. Lastly, we demonstrated a step-wise improvement in lycopene production from shake-flasks to a 5-L fermenter using the strain with GAL80 intact. Altogether, our study represents novel findings on more effective crt genes from Sp. pararoseus over the previously reported benchmark genes and their potential applications in scale-up lycopene production.
Collapse
Affiliation(s)
- Akaraphol Watcharawipas
- National Center for Genetic Engineering and Biotechnology, 113 Thailand Science Park, Paholyothin Road, Klong 1, Klong Luang, Pathumthani 12120, Thailand
| | - Kitisak Sansatchanon
- National Center for Genetic Engineering and Biotechnology, 113 Thailand Science Park, Paholyothin Road, Klong 1, Klong Luang, Pathumthani 12120, Thailand
| | - Chitwadee Phithakrotchanakoon
- National Center for Genetic Engineering and Biotechnology, 113 Thailand Science Park, Paholyothin Road, Klong 1, Klong Luang, Pathumthani 12120, Thailand
| | - Sutipa Tanapongpipat
- National Center for Genetic Engineering and Biotechnology, 113 Thailand Science Park, Paholyothin Road, Klong 1, Klong Luang, Pathumthani 12120, Thailand
| | - Weerawat Runguphan
- National Center for Genetic Engineering and Biotechnology, 113 Thailand Science Park, Paholyothin Road, Klong 1, Klong Luang, Pathumthani 12120, Thailand
| | - Kanokarn Kocharin
- National Center for Genetic Engineering and Biotechnology, 113 Thailand Science Park, Paholyothin Road, Klong 1, Klong Luang, Pathumthani 12120, Thailand
| |
Collapse
|
|
19
|
Impairment of carotenoid biosynthesis through CAR1 gene mutation results in CoQ 10, sterols, and phytoene accumulation in Rhodotorula mucilaginosa. Appl Microbiol Biotechnol 2021; 106:317-327. [PMID: 34910239 DOI: 10.1007/s00253-021-11673-5] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/11/2021] [Revised: 10/26/2021] [Accepted: 10/28/2021] [Indexed: 10/19/2022]
Abstract
Red yeasts, mainly included in the genera Rhodotorula, Rhodosporidiobolus, and Sporobolomyces, are renowned biocatalysts for the production of a wide range of secondary metabolites of commercial interest, among which lipids, carotenoids, and other isoprenoids. The production of all these compounds is tightly interrelated as they share acetyl-CoA and the mevalonate pathway as common intermediates. Here, T-DNA insertional mutagenesis was applied to the wild type strain C2.5t1 of Rhodotorula mucilaginosa for the isolation of albino mutants with impaired carotenoids biosynthesis. The rationale behind this approach was that a blockage in carotenoid biosynthetic pathway could divert carbon flux toward the production of lipids and/or other molecules deriving from terpenoid precursors. One characterized albino mutant, namely, strain W4, carries a T-DNA insertion in the CAR1 gene coding for phytoene desaturase. When cultured in glycerol-containing medium, W4 strain showed significant decreases in cell density and fatty acids content in respect to the wild type strain. Conversely, it reached significantly higher productions of phytoene, CoQ10, and sterols. These were supported by an increased expression of CAR2 gene that codes for phytoene synthase/lycopene cyclase. Thus, in accordance with the starting hypothesis, the impairment of carotenoids biosynthesis can be explored to pursue the biotechnological exploitation of red yeasts for enhanced production of secondary metabolites with several commercial applications. KEY POINTS: • The production of lipids, carotenoids, and other isoprenoids is tightly interrelated. • CAR1 gene mutation results in the overproduction of phytoene, CoQ10, and sterols. • Albino mutants are promising tools for the production of secondary metabolites.
Collapse
|
|
20
|
Abeln F, Chuck CJ. The history, state of the art and future prospects for oleaginous yeast research. Microb Cell Fact 2021; 20:221. [PMID: 34876155 PMCID: PMC8650507 DOI: 10.1186/s12934-021-01712-1] [Citation(s) in RCA: 54] [Impact Index Per Article: 13.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/29/2021] [Accepted: 11/23/2021] [Indexed: 12/25/2022] Open
Abstract
Lipid-based biofuels, such as biodiesel and hydroprocessed esters, are a central part of the global initiative to reduce the environmental impact of the transport sector. The vast majority of production is currently from first-generation feedstocks, such as rapeseed oil, and waste cooking oils. However, the increased exploitation of soybean oil and palm oil has led to vast deforestation, smog emissions and heavily impacted on biodiversity in tropical regions. One promising alternative, potentially capable of meeting future demand sustainably, are oleaginous yeasts. Despite being known about for 143 years, there has been an increasing effort in the last decade to develop a viable industrial system, with currently around 100 research papers published annually. In the academic literature, approximately 160 native yeasts have been reported to produce over 20% of their dry weight in a glyceride-rich oil. The most intensively studied oleaginous yeast have been Cutaneotrichosporon oleaginosus (20% of publications), Rhodotorula toruloides (19%) and Yarrowia lipolytica (19%). Oleaginous yeasts have been primarily grown on single saccharides (60%), hydrolysates (26%) or glycerol (19%), and mainly on the mL scale (66%). Process development and genetic modification (7%) have been applied to alter yeast performance and the lipids, towards the production of biofuels (77%), food/supplements (24%), oleochemicals (19%) or animal feed (3%). Despite over a century of research and the recent application of advanced genetic engineering techniques, the industrial production of an economically viable commodity oil substitute remains elusive. This is mainly due to the estimated high production cost, however, over the course of the twenty-first century where climate change will drastically change global food supply networks and direct governmental action will likely be levied at more destructive crops, yeast lipids offer a flexible platform for localised, sustainable lipid production. Based on data from the large majority of oleaginous yeast academic publications, this review is a guide through the history of oleaginous yeast research, an assessment of the best growth and lipid production achieved to date, the various strategies employed towards industrial production and importantly, a critical discussion about what needs to be built on this huge body of work to make producing a yeast-derived, more sustainable, glyceride oil a commercial reality.
Collapse
Affiliation(s)
- Felix Abeln
- Department of Chemical Engineering, University of Bath, Bath, BA2 7AY, UK.
- Centre for Sustainable and Circular Technologies, University of Bath, Bath, BA2 7AY, UK.
| | | |
Collapse
|
|
21
|
Martín-Hernández GC, Müller B, Chmielarz M, Brandt C, Hölzer M, Viehweger A, Passoth V. Chromosome-level genome assembly and transcriptome-based annotation of the oleaginous yeast Rhodotorula toruloides CBS 14. Genomics 2021; 113:4022-4027. [PMID: 34648882 DOI: 10.1016/j.ygeno.2021.10.006] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/08/2021] [Revised: 08/26/2021] [Accepted: 10/07/2021] [Indexed: 11/30/2022]
Abstract
Rhodotorula toruloides is an oleaginous yeast with high biotechnological potential. In order to understand the molecular physiology of lipid synthesis in R. toruloides and to advance metabolic engineering, a high-resolution genome is required. We constructed a genome draft of R. toruloides CBS 14, using a hybrid assembly approach, consisting of short and long reads generated by Illumina and Nanopore sequencing, respectively. The genome draft consists of 23 contigs and 3 scaffolds, with a N50 length of 1,529,952 bp, thus largely representing chromosomal organization. The total size of the genome is 20,534,857 bp and the overall GC content is 61.83%. Transcriptomic data from different growth conditions was used to aid species-specific gene annotation. We annotated 9464 genes and identified 11,691 transcripts. Furthermore, we demonstrated the presence of a potential plasmid, an extrachromosomal circular structure of about 11 kb with a copy number about three times as high as the other chromosomes.
Collapse
Affiliation(s)
| | - Bettina Müller
- Department of Molecular Sciences, Swedish University of Agricultural Sciences, Uppsala, Sweden
| | - Mikołaj Chmielarz
- Department of Molecular Sciences, Swedish University of Agricultural Sciences, Uppsala, Sweden
| | - Christian Brandt
- nanozoo GmbH, Leipzig, Germany; Institute for Infectious Diseases and Infection Control, Jena University Hospital, Jena, Germany
| | - Martin Hölzer
- nanozoo GmbH, Leipzig, Germany; RNA Bioinformatics and High-Throughput Analysis, Friedrich Schiller University Jena, Jena, Germany
| | - Adrian Viehweger
- nanozoo GmbH, Leipzig, Germany; Department of Medical Microbiology, University Hospital Leipzig, Germany
| | - Volkmar Passoth
- Department of Molecular Sciences, Swedish University of Agricultural Sciences, Uppsala, Sweden.
| |
Collapse
|
|
22
|
Chen QH, Wu BK, Pan D, Sang LX, Chang B. Beta-carotene and its protective effect on gastric cancer. World J Clin Cases 2021; 9:6591-6607. [PMID: 34447808 PMCID: PMC8362528 DOI: 10.12998/wjcc.v9.i23.6591] [Citation(s) in RCA: 28] [Impact Index Per Article: 7.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 01/12/2021] [Revised: 05/16/2021] [Accepted: 06/22/2021] [Indexed: 02/06/2023] Open
Abstract
Beta-carotene is an important natural pigment that is very beneficial to human health. It is widely found in vegetables and fruits. The three main functions are antioxidant effects, cell gap junction-related functions and immune-related functions. Because of its diverse functions, beta-carotene is believed to prevent and treat many chronic diseases. Gastric cancer is one of the most important diseases it can treat. Gastric cancer is a type of cancer with a high incidence. Its etiology varies, and the pathogenesis is complex. Gastric cancer seriously affects human health. The role of beta-carotene, a natural nutrient, in gastric cancer has been explored by many researchers, including molecular mechanisms and epidemiological studies. Molecular studies have mainly focused on oxidative stress, cell cycle, signal transduction pathways and immune-related mechanisms of beta-carotene in gastric cancer. Many epidemiological surveys and cohort studies of patients with gastric cancer have been conducted, and the results of these epidemiological studies vary due to the use of different research methods and analysis of different regions. This paper will summarize the results of these studies, mainly in terms of molecular mechanisms and epidemiological research results, which will provide a systematic basis for future studies of the treatment and prognosis of gastric cancer. This paper will help researchers identify new research directions.
Collapse
Affiliation(s)
- Qian-Hui Chen
- Department of Intensive Care Unit, First Affiliated Hospital of China Medical University, Shenyang 110001, Liaoning Province, China
| | - Bao-Kang Wu
- Department of General Surgery, Shengjing Hospital of China Medical University, Shenyang 110004, Liaoning Province, China
| | - Dan Pan
- Department of Geriatrics, First Affiliated Hospital of China Medical University, Shenyang 110001, Liaoning Province, China
| | - Li-Xuan Sang
- Department of Geriatrics, First Affiliated Hospital of China Medical University, Shenyang 110001, Liaoning Province, China
| | - Bing Chang
- Department of Gastroenterology, First Affiliated Hospital of China Medical University, Shenyang 110001, Liaoning Province, China
| |
Collapse
|
|
23
|
Byrtusová D, Szotkowski M, Kurowska K, Shapaval V, Márová I. Rhodotorula kratochvilovae CCY 20-2-26-The Source of Multifunctional Metabolites. Microorganisms 2021; 9:1280. [PMID: 34208382 PMCID: PMC8231246 DOI: 10.3390/microorganisms9061280] [Citation(s) in RCA: 11] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/29/2021] [Revised: 06/01/2021] [Accepted: 06/08/2021] [Indexed: 01/23/2023] Open
Abstract
Multifunctional biomass is able to provide more than one valuable product, and thus, it is attractive in the field of microbial biotechnology due to its economic feasibility. Carotenogenic yeasts are effective microbial factories for the biosynthesis of a broad spectrum of biomolecules that can be used in the food and feed industry and the pharmaceutical industry, as well as a source of biofuels. In the study, we examined the effect of different nitrogen sources, carbon sources and CN ratios on the co-production of intracellular lipids, carotenoids, β-glucans and extracellular glycolipids. Yeast strain R. kratochvilovae CCY 20-2-26 was identified as the best co-producer of lipids (66.7 ± 1.5% of DCW), exoglycolipids (2.42 ± 0.08 g/L), β-glucan (11.33 ± 1.34% of DCW) and carotenoids (1.35 ± 0.11 mg/g), with a biomass content of 15.2 ± 0.8 g/L, by using the synthetic medium with potassium nitrate and mannose as a carbon source. It was shown that an increased C/N ratio positively affected the biomass yield and production of lipids and β-glucans.
Collapse
Affiliation(s)
- Dana Byrtusová
- Faculty of Science and Technology, Norwegian University of Life Sciences, P.O. Box 5003, 1432 Ås, Norway; (D.B.); (V.S.)
- Faculty of Chemistry, Brno University of Technology, Purkyňova 464/118, 612 00 Brno, Czech Republic; (M.S.); (K.K.)
| | - Martin Szotkowski
- Faculty of Chemistry, Brno University of Technology, Purkyňova 464/118, 612 00 Brno, Czech Republic; (M.S.); (K.K.)
| | - Klára Kurowska
- Faculty of Chemistry, Brno University of Technology, Purkyňova 464/118, 612 00 Brno, Czech Republic; (M.S.); (K.K.)
| | - Volha Shapaval
- Faculty of Science and Technology, Norwegian University of Life Sciences, P.O. Box 5003, 1432 Ås, Norway; (D.B.); (V.S.)
| | - Ivana Márová
- Faculty of Chemistry, Brno University of Technology, Purkyňova 464/118, 612 00 Brno, Czech Republic; (M.S.); (K.K.)
| |
Collapse
|
|
24
|
Chattopadhyay A, Maiti MK. Lipid production by oleaginous yeasts. ADVANCES IN APPLIED MICROBIOLOGY 2021; 116:1-98. [PMID: 34353502 DOI: 10.1016/bs.aambs.2021.03.003] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 02/06/2023]
Abstract
Microbial lipid production has been studied extensively for years; however, lipid metabolic engineering in many of the extraordinarily high lipid-accumulating yeasts was impeded by inadequate understanding of the metabolic pathways including regulatory mechanisms defining their oleaginicity and the limited genetic tools available. The aim of this review is to highlight the prominent oleaginous yeast genera, emphasizing their oleaginous characteristics, in conjunction with diverse other features such as cheap carbon source utilization, withstanding the effect of inhibitory compounds, commercially favorable fatty acid composition-all supporting their future development as economically viable lipid feedstock. The unique aspects of metabolism attributing to their oleaginicity are accentuated in the pretext of outlining the various strategies successfully implemented to improve the production of lipid and lipid-derived metabolites. A large number of in silico data generated on the lipid accumulation in certain oleaginous yeasts have been carefully curated, as suggestive evidences in line with the exceptional oleaginicity of these organisms. The different genetic elements developed in these yeasts to execute such strategies have been scrupulously inspected, underlining the major types of newly-found and synthetically constructed promoters, transcription terminators, and selection markers. Additionally, there is a plethora of advanced genetic toolboxes and techniques described, which have been successfully used in oleaginous yeasts in the recent years, promoting homologous recombination, genome editing, DNA assembly, and transformation at remarkable efficiencies. They can accelerate and effectively guide the rational designing of system-wide metabolic engineering approaches pinpointing the key targets for developing industrially suitable yeast strains.
Collapse
Affiliation(s)
- Atrayee Chattopadhyay
- Department of Biotechnology, Indian Institute of Technology Kharagpur, Kharagpur, India
| | - Mrinal K Maiti
- Department of Biotechnology, Indian Institute of Technology Kharagpur, Kharagpur, India.
| |
Collapse
|
|
25
|
Chattopadhyay A, Mitra M, Maiti MK. Recent advances in lipid metabolic engineering of oleaginous yeasts. Biotechnol Adv 2021; 53:107722. [PMID: 33631187 DOI: 10.1016/j.biotechadv.2021.107722] [Citation(s) in RCA: 31] [Impact Index Per Article: 7.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/17/2020] [Revised: 02/15/2021] [Accepted: 02/15/2021] [Indexed: 01/12/2023]
Abstract
With the increasing demand to develop a renewable and sustainable biolipid feedstock, several species of non-conventional oleaginous yeasts are being explored. Apart from the platform oleaginous yeast Yarrowia lipolytica, the understanding of metabolic pathway and, therefore, exploiting the engineering prospects of most of the oleaginous species are still in infancy. However, in the past few years, enormous efforts have been invested in Rhodotorula, Rhodosporidium, Lipomyces, Trichosporon, and Candida genera of yeasts among others, with the rapid advancement of engineering strategies, significant improvement in genetic tools and techniques, generation of extensive bioinformatics and omics data. In this review, we have collated these recent progresses to make a detailed and insightful summary of the major developments in metabolic engineering of the prominent oleaginous yeast species. Such a comprehensive overview would be a useful resource for future strain improvement and metabolic engineering studies for enhanced production of lipid and lipid-derived chemicals in oleaginous yeasts.
Collapse
Affiliation(s)
- Atrayee Chattopadhyay
- Department of Biotechnology, Indian Institute of Technology Kharagpur, Kharagpur 721302, India
| | - Mohor Mitra
- Department of Biotechnology, Indian Institute of Technology Kharagpur, Kharagpur 721302, India
| | - Mrinal K Maiti
- Department of Biotechnology, Indian Institute of Technology Kharagpur, Kharagpur 721302, India.
| |
Collapse
|
|
26
|
Parasar DP, Ramakrishnan E, Kabilan S, Kotoky J, Sarma HK. Characterization of β-Cryptoxanthin and Other Carotenoid Derivatives from Rhodotorula taiwanensis, A Novel Yeast Isolated from Traditional Starter Culture of Assam. Chem Biodivers 2020; 17:e2000198. [PMID: 33094905 DOI: 10.1002/cbdv.202000198] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/18/2020] [Accepted: 10/20/2020] [Indexed: 12/31/2022]
Abstract
The present investigation was intended to characterize pigments for the first time from Rhodotorula taiwanensis (LC011412) yeast isolated from the ethic fermentation starter culture source meant to evaluate its carotenoid contents for beneficial applications. The pigments were extracted by an optimized solvent system, purified by flash chromatography and were identified by TLC and UV/VIS spectroscopy. The absorbance spectra confirmed the presence of β-carotene, β-cryptoxanthin, torulene and torularhodin that showed maximum absorbance (λmax ) within the ranges. The fractions were further characterized by LC/MS and analyzed through FT-IR and NMR for structure elucidation. Spectral analyses also confirmed the presence of the compounds mentioned above. These compounds promise great commercial value and could be useful for large scale production anticipated for potential applications in food, nutraceutical, pharmaceutical and cosmetic sectors. It is pertinent that the characterized carotenoid pigments from the isolate have incredible prospects in industrial applications which require profound attention.
Collapse
Affiliation(s)
- Deep Prakash Parasar
- Microbial Communication and Fungal Biology Group, Department of Biotechnology, Gauhati University, 781014, Guwahati, Assam, India.,Department of Biotechnology, Assam Down Town University, 781026, Panikhaiti, Guwahati, Assam, India
| | - Elancheran Ramakrishnan
- Drug Discovery Lab, Department of Chemistry, Annamalai University, Annamalai Nagar, 608002, Tamil Nadu, India
| | - Senthamaraikannan Kabilan
- Drug Discovery Lab, Department of Chemistry, Annamalai University, Annamalai Nagar, 608002, Tamil Nadu, India
| | - Jibon Kotoky
- Drug Discovery Laboratory, Life Science Division, Institute of Advanced Study in Science and Technology (IASST), Paschim Boragaon, 781035, Guwahati, Assam, India
| | - Hridip Kumar Sarma
- Microbial Communication and Fungal Biology Group, Department of Biotechnology, Gauhati University, 781014, Guwahati, Assam, India
| |
Collapse
|
|
27
|
Liu X, Li Y, Yu Z, Liu X, Hardie WJ, Huang M. Screening and characterisation of β-glucosidase production strains from Rosa roxburghii Tratt. INTERNATIONAL JOURNAL OF FOOD ENGINEERING 2020. [DOI: 10.1515/ijfe-2020-0152] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 11/15/2022]
Abstract
Abstract
The β-glucosidase properties from one yeast isolate identified as Wickerhamomyces anomalus C4 were characterised. The β-glucosidase activity of W. anomalus C4 was 41.83 ± 0.25 mU/mL, and the optimum temperature and pH were 40 °C and 5.0, respectively. The glucose, 10% v/v of ethanol and 10 mmol/L of Cu2+ inhibited the β-glycosidases activities. The isolate W. anomalus C4 had a stronger alcohol metabolism capacity than commercial Saccharomyces cerevisiae X16. Besides, fermentation with W. anomalus C4 alone and co-fermentations with S. cerevisiae X16 and W. anomalus C4 reduced the volatile acids content and the sourness value compared to S. cerevisiae X16 control. Moreover, inoculation with W. anomalus C4 could enhance volatile aroma richness and complexity of Rosa roxburghii wines, regardless of type or amount thereof. Therefore, the R. roxburghii native yeast isolate W. anomalus C4 may have some application potentials for R. roxburghii wine-making.
Collapse
Affiliation(s)
- Xiaozhu Liu
- Guizhou Institute of Technology , Guiyang 550003 , Guizhou , China
| | - Yinfeng Li
- Guizhou Institute of Technology , Guiyang 550003 , Guizhou , China
| | - Zhihai Yu
- Guizhou Institute of Technology , Guiyang 550003 , Guizhou , China
| | - Xiaohui Liu
- Guizhou Institute of Technology , Guiyang 550003 , Guizhou , China
| | - William James Hardie
- Guizhou Institute of Technology , Guiyang 550003 , Guizhou , China
- Changzhou University , Changzhou 213000 , Jiangsu , China
| | - Mingzheng Huang
- Guizhou Institute of Technology , Guiyang 550003 , Guizhou , China
| |
Collapse
|
|
28
|
Carotenoid-producing yeasts: Identification and Characteristics of Environmental Isolates with a Valuable Extracellular Enzymatic Activity. Microorganisms 2019; 7:microorganisms7120653. [PMID: 31817221 PMCID: PMC6956281 DOI: 10.3390/microorganisms7120653] [Citation(s) in RCA: 17] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/31/2019] [Revised: 12/03/2019] [Accepted: 12/03/2019] [Indexed: 12/03/2022] Open
Abstract
Sixteen cold-adapted reddish-pigmented yeast strains were obtained from environmental samples. According to the PCR-based detection of classical yeast markers combined with phylogenetic studies, the yeasts belong mainly to the genera Rhodotorula, Sporobolomyces and Cystobasidium, all within the subphylum Pucciniomycotina. All strains produced carotenoids within a 0.25–10.33 mg/L range under non-optimized conditions. Noteworthily, among them, representatives of the Cystobasidium genus were found; of particular value are the strains C. laryngis and C. psychroaquaticum, poorly described in the literature to date. Interestingly, carotenoid production with representatives of Cystobasidium was improved 1.8- to 10-fold at reduced temperature. As expected, most of the isolated yeasts biosynthesized extracellular lipases, but within them also one proteolytic and four cellulolytic strains were revealed. We succeeded in isolating strain Cystofilobasidium macerans WUT145 with extraordinarily high cellulolytic activity at 22°C (66.23 ± 0.15 µmol/mg protein·min) that is described here for the first time. Consequently, a set of yeasts capable of producing both carotenoids and extracellular enzymes was identified. Taking into account those abilities, the strains might be applicable for a development of carotenoids production on an agro-industrial waste, e.g., lignocellulose.
Collapse
|
|
29
|
Tiukova IA, Prigent S, Nielsen J, Sandgren M, Kerkhoven EJ. Genome‐scale model of
Rhodotorula toruloides
metabolism. Biotechnol Bioeng 2019; 116:3396-3408. [DOI: 10.1002/bit.27162] [Citation(s) in RCA: 33] [Impact Index Per Article: 5.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/01/2019] [Revised: 08/08/2019] [Accepted: 09/05/2019] [Indexed: 12/31/2022]
Affiliation(s)
- Ievgeniia A. Tiukova
- Systems and Synthetic Biology, Department of Biology and Biological EngineeringChalmers University of TechnologyGothenburg Sweden
- Department of Molecular SciencesSwedish University of Agricultural SciencesUppsala Sweden
| | | | - Jens Nielsen
- Systems and Synthetic Biology, Department of Biology and Biological EngineeringChalmers University of TechnologyGothenburg Sweden
| | - Mats Sandgren
- Department of Molecular SciencesSwedish University of Agricultural SciencesUppsala Sweden
| | - Eduard J. Kerkhoven
- Systems and Synthetic Biology, Department of Biology and Biological EngineeringChalmers University of TechnologyGothenburg Sweden
| |
Collapse
|
|
30
|
Gong G, Liu L, Zhang X, Tan T. Comparative evaluation of different carbon sources supply on simultaneous production of lipid and carotene of Rhodotorula glutinis with irradiation and the assessment of key gene transcription. BIORESOURCE TECHNOLOGY 2019; 288:121559. [PMID: 31152958 DOI: 10.1016/j.biortech.2019.121559] [Citation(s) in RCA: 18] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 04/16/2019] [Revised: 05/22/2019] [Accepted: 05/23/2019] [Indexed: 06/09/2023]
Abstract
To investigate the feasibility of simultaneously enhancing lipid and carotene production by irradiation with different carbon sources, a strategy by controlling the carbon sources supply were selected to culture Rhodotorula glutinis under the irradiation condition. The results demonstrated that the irradiation indeed enhanced cell growth, lipid and carotene production with different carbon sources supply. Besides, the fatty acids profiling as revealed by more unsaturated fatty acids (mainly C16:1, C18:2 and C18:3) and less saturated fatty acids (C18:0, C22:0 and C24:0) were found during the process of irradiation. Compared with the control, the increase of the transcription levels in genes connected with substrates assimilation, lipid production and carotene accumulation were observed under the irradiation condition. The results suggest the possibility of using irradiation as an effective strategy to increase the production of both lipid and carotene with the controlled carbon sources supply.
Collapse
Affiliation(s)
- Guiping Gong
- Beijing Key Lab of Bioprocess, National Energy R&D Center for Biorefinery, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, PR China
| | - Luo Liu
- Beijing Key Lab of Bioprocess, National Energy R&D Center for Biorefinery, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, PR China
| | - Xu Zhang
- Beijing Key Lab of Bioprocess, National Energy R&D Center for Biorefinery, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, PR China.
| | - Tianwei Tan
- Beijing Key Lab of Bioprocess, National Energy R&D Center for Biorefinery, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, PR China
| |
Collapse
|
|
31
|
Arhar S, Natter K. Common aspects in the engineering of yeasts for fatty acid- and isoprene-based products. Biochim Biophys Acta Mol Cell Biol Lipids 2019; 1864:158513. [PMID: 31465888 DOI: 10.1016/j.bbalip.2019.08.009] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/10/2019] [Revised: 06/26/2019] [Accepted: 08/20/2019] [Indexed: 11/18/2022]
Abstract
The biosynthetic pathways for most lipophilic metabolites share several common principles. These substances are built almost exclusively from acetyl-CoA as the donor for the carbon scaffold and NADPH is required for the reductive steps during biosynthesis. Due to their hydrophobicity, the end products are sequestered into the same cellular compartment, the lipid droplet. In this review, we will summarize the efforts in the metabolic engineering of yeasts for the production of two major hydrophobic substance classes, fatty acid-based lipids and isoprenoids, with regard to these common aspects. We will compare and discuss the results of genetic engineering strategies to construct strains with enhanced synthesis of the precursor acetyl-CoA and with modified redox metabolism for improved NADPH supply. We will also discuss the role of the lipid droplet in the storage of the hydrophobic product and review the strategies to either optimize this organelle for higher capacity or to achieve excretion of the product into the medium.
Collapse
Affiliation(s)
- Simon Arhar
- Institute of Molecular Biosciences, NAWI Graz, University of Graz, Humboldtstrasse 50/II, 8010 Graz, Austria
| | - Klaus Natter
- Institute of Molecular Biosciences, NAWI Graz, University of Graz, Humboldtstrasse 50/II, 8010 Graz, Austria.
| |
Collapse
|
|
32
|
Schultz JC, Cao M, Zhao H. Development of a CRISPR/Cas9 system for high efficiency multiplexed gene deletion in Rhodosporidium toruloides. Biotechnol Bioeng 2019; 116:2103-2109. [PMID: 31038202 DOI: 10.1002/bit.27001] [Citation(s) in RCA: 37] [Impact Index Per Article: 6.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/07/2019] [Revised: 04/11/2019] [Accepted: 04/18/2019] [Indexed: 01/06/2023]
Abstract
The oleaginous yeast Rhodosporidium toruloides is considered a promising candidate for production of chemicals and biofuels thanks to its ability to grow on lignocellulosic biomass, and its high production of lipids and carotenoids. However, efforts to engineer this organism are hindered by a lack of suitable genetic tools. Here we report the development of a CRISPR/Cas9 system for genome editing in R. toruloides based on a fusion 5S rRNA-tRNA promoter for guide RNA (gRNA) expression, capable of greater than 95% gene knockout for various genetic targets. Additionally, multiplexed double-gene knockout mutants were obtained using this method with an efficiency of 78%. This tool can be used to accelerate future metabolic engineering work in this yeast.
Collapse
Affiliation(s)
- J Carl Schultz
- Department of Chemical and Biomolecular Engineering, Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois
| | - Mingfeng Cao
- Department of Chemical and Biomolecular Engineering, Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois
| | - Huimin Zhao
- Department of Chemical and Biomolecular Engineering, Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois
- Departments of Chemistry, Biochemistry, and Bioengineering, University of Illinois at Urbana-Champaign, Urbana, Illinois
| |
Collapse
|
|
33
|
Jinendiran S, Dileep Kumar BS, Dahms HU, Arulanandam CD, Sivakumar N. Optimization of submerged fermentation process for improved production of β-carotene by Exiguobacterium acetylicum S01. Heliyon 2019; 5:e01730. [PMID: 31193511 PMCID: PMC6535579 DOI: 10.1016/j.heliyon.2019.e01730] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/20/2019] [Revised: 04/15/2019] [Accepted: 05/10/2019] [Indexed: 11/23/2022] Open
Abstract
Carotenoids are natural pigments with substantial applications in nutraceutical, pharmaceutical, and food industries. In this study, optimization of the fermentation process for enhancement of β-carotene and biomass production by Exiguobacterium acetylicum S01 was achieved by employing statistical designs including the Placket-Burman design (PBD) and response surface methodology (RSM). Among the seven variables investigated by two levels in PBD, glucose, peptone, pH and temperature were indicated as crucial variables (p < 0.0001) for β-carotene and biomass productivity. Response surface methodology was further applied to evaluate the optimal concentrations of these four variables for maximum β-carotene and biomass productivity. The optimized medium contained glucose 1.4 g/L, peptone 26.5 g/L, pH 8.5, and temperature 30 °C, respectively. A significant increase in β-carotene (40.32 ± 2.55 mg/L) and biomass (2.19 ± 0.10 g/L) productivities in E. acetylicum S01 were achieved by using RSM, which was 3.47-fold and 2.36-fold higher in the optimized medium compared to the un-optimized medium. Further, the optimum fermentation condition in the 5-L bioreactor was achieved a maximal β-carotene yield of 107.22 ± 5.78 mg/L within 96 h. Moreover, the expression levels of carotenoid biosynthetic genes (phytoene desaturase (CrtI) and phytoene synthase (CrtB)) were up-regulated (2.89-fold and 3.71-fold) in E. acetylicum under the optimized medium conditions. Overall, these results suggest that E. acetylicum S01 can be used as a promising microorganism for the commercial production of β-carotene.
Collapse
Affiliation(s)
- Sekar Jinendiran
- Department of Molecular Microbiology, School of Biotechnology, Madurai Kamaraj University, Madurai, 625021, India
| | - B S Dileep Kumar
- Agro-Processing and Technology Division, CSIR-National Institute for Interdisciplinary Science and Technology, Thiruvananthapuram, 695019, India
| | - Hans-Uwe Dahms
- Department of Biomedical Science and Environmental Biology, Kaohsiung Medical University, Kaohsiung, 80708, Taiwan.,Department of Marine Biotechnology and Bioresources, National Sun Yat-sen University, Kaohsiung, 80424, Taiwan
| | - Charli Deepak Arulanandam
- Department of Biomedical Science and Environmental Biology, Kaohsiung Medical University, Kaohsiung, 80708, Taiwan
| | - Natesan Sivakumar
- Department of Molecular Microbiology, School of Biotechnology, Madurai Kamaraj University, Madurai, 625021, India
| |
Collapse
|