Frontiers in Bioengineering and Biotechnology
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2022
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IF 6.064
Zhang Yue,Zhang Sufang,Chu Yadong,Zhang Qi,Zhou Renhui,Yu Di,Wang Shuang,Lyu Liting,Xu Guowang,Zhao Zongbao Kent
The basidiomycetous yeast Rhodosporidium toruloides (R. toruloides) is an excellent producer for neutral lipids, including triacylglycerols (TAG). Partially because genetic tools for this yeast were less developed, limited efforts were shown to explore its capacity for the production of higher-value lipids such as diacylglycerols (DAG). Here, four genes linked to the interconversion between DAG and TAG were manipulated to promote the production of DAG and free fatty acids (FFA). Among them, three TAG synthesis-related genes, DGA1, LRO1, and ARE1, were down-regulated successively via the RNA interference technology, and an endogenous TAG lipase encoded by TGL5 was fused with LDP1 and over-expressed to convert TAG into DAG and FFA. Results showed that those engineered R. toruloides strains grew normally under nutrient-rich conditions but notably slower than the parental strain NP11 in the lipid production stage. When cultivated in nitrogen-limited media, engineered strains were able to produce total lipids with improved contents of DAG and FFA by up to two-fold and three-fold, respectively. Further correlation analysis between lipid composition and cell density indicated that the formation of TAG correlated positively with cell growth; however, other lipids including DAG did negatively. This study offered valuable information and strains to engineer R. toruloides for advanced production of fatty acid derivatives.
研究领域:
微生物学
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遗传学
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代谢工程
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DOI
PubMed
高影响因子
Frontiers in Bioengineering and Biotechnology
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2021
·
IF 5.89
Lyu Liting,Chu Yadong,Zhang Sufang,Zhang Yue,Huang Qitian,Wang Shuang,Zhao Zongbao K.
Conversion of lignocellulosic biomass into lipids and related chemicals has attracted much attention in the past two decades, and the oleaginous yeast Rhodosporidium toruloides has been widely used in this area. While R. toruloides species naturally have physiological advantages in terms of substrate utilization, lipid accumulation, and inhibitor resistance, reduced lipid production and cell growth are noticed when biomass hydrolysates are used as feedstocks. To improve the robustness of R. toruloides, here, we devised engineered strains by overexpressing genes responsible for phenolic compound degradation. Specifically, gene expression cassettes of the manganese peroxidase gene (MNP) and versatile peroxidase gene (VP) were constructed and integrated into the genome of R. toruloides NP11. A series of engineered strains were evaluated for lipid production in the presence of typical phenolic inhibitors. The results showed that R. toruloides strains with proper expression of MNP or VP indeed grew faster in the presence of vanillin and 5-hydroxymethylfurfural than the parental strain. When cultivated in concentrated mode biomass hydrolysates, the strain VP18 had improved performance as the cell mass and lipid content increased by 30% and 25%, respectively. This study provides more robust oleaginous yeast strains for microbial lipid production from lignocellulosic biomass, and similar efforts may be used to devise more advanced lipid producers.
研究领域:
生物技术
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微生物学
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DOI
PubMed
高影响因子
Food & Function
·
2023
·Vol. 14
·期 14
·6363-6375
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IF 6.1
Mengyang Liu,Haitao Wang,Xu Yan,Sufang Zhang,Chaofan Ji,Yingxi Chen,Beiwei Zhu,Xinping Lin
A carotenoid production strain Rhodosporidium toruloides NP11 and its mutant strain R. toruloides A1-15 were studied under chemostat nitrogen-limited cultivation. Multi-omics analysis (metabolomics, lipidomics and transcriptomics) was used to investigate the different mechanisms of torularhodin accumulation between NP11 and A1-15. The results showed that the carotenoid synthesis pathway was significantly enhanced in A1-15 compared to NP11 under nitrogen limitation, due to the significant increase of torularhodin. Under nitrogen-limited conditions, higher levels of β-oxidation were present in A1-15 compared to those in NP11, which provided sufficient precursors for carotenoid synthesis. In addition, ROS stress accelerated the intracellular transport of iron ions, promoted the expression of CRTI and CRTY genes, and reduced the transcript levels of FNTB1 and FNTB2 in the bypass pathway, and these factors may be responsible for the regulation of high torularhodin production in A1-15. This study provided insights into the selective production of torularhodin.
研究领域:
微生物学
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代谢组学
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脂质组学
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转录组学
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生物技术
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DOI
PubMed
高影响因子
BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY
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2020
·Vol. 68
·期 1
·71-81
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IF 1.638
Zeping Sun,Jing Lv,Chaofan Ji,Huipeng Liang,Shengjie Li,Zhaoxia Yang,Wenhuan Xu,Sufang Zhang,Xinping Lin
Rhodosporidium toruloides has been reported as a potential biotechnological microorganism to produce carotenoids. The most commonly used molecular and genetic manipulation methods based on Agrobacterium -mediated transformation (ATMT). However, this method was of relatively lower transformation efficiency. In this study, we optimized the ATMT method for R. toruloides on account of the promoter on T-DNA, the ratio of A. tumefaciens to R. toruloides NP11, acetosyringone concentration, cocultivation temperature and time, and a transformation efficiency of 2,369 cells per 10 5 recipient cells was obtained and was 24 times as that of the previous report. With this optimized method, four redder mutants and four yellower mutants were selected out with torularhodin and β-carotene production preference, respectively. The highest torularhodin production was 1,638.15 µg/g dry cell weight in A1-13. The yellower mutants were found to divert the metabolic flux from torularhodin and torulene to γ-carotene and β-carotene, and the proportion of γ-carotene and β-carotene were all over 92%. TAIL-PCR was carried out to found T-DNA insertion in these mutants, and insertion hotspot was found. RT-qPCR results showed that CTA 1 genes in these mutants were closely related to the synthesis of total carotenoids, especially torularhodin, and was a potenial metabolic engineering site in the future.
研究领域:
微生物学
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分子生物学
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生物技术
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PubMed
高影响因子
RSC Advances
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2014
·Vol. 4
·期 66
·34944-34949
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IF 3.708
Jintao Lin,Shuangyue Li,Mingzhong Sun,Cuili Zhang,Wenbo Yang,Zehui Zhang,Xiaojie Li,Shao Li
The economical production of biodiesel from lignocellulose hydrolysates requires a robust D-xylose fermentation path. The present work found that Lipomyces starkeyi AS 2.1560 and Rhodosporidium toruloides AS 2.1389 could produce micro-lipid effectively from D-xylose solution by a two-stage culture mode. Results demonstrated that both L. starkeyi and R. toruloides could achieve high lipid production in xylose solution. High cell density fermentation was carried out by a feed-batch fermentation mode. After fermentation for 48 h in a bioreactor using unsterile xylose solution by L. starkeyi, the biomass, lipid and lipid content reached 97.4 g L−1, 63.9 g L−1, and 65.5%, respectively. Accordingly, the lipid productivity reached 1.3 g L−1 h−1 in the second stage. And the overall lipid productivity was 0.55 g L−1 h−1. Therefore, L. starkeyi and R. toruloides are highly effective D-xylose-fermenting oleaginous yeasts for lipid production in xylose solution. The two-stage culture method should provide a new opportunity to make the industrial production of biodiesel possible in the future.
研究领域:
微生物学
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生物技术
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可再生能源
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DOI
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2019
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IF 3.571
Ruiqi Bao,Ning Gao,Jing Lv,Chaofan Ji,Huipeng Liang,Shengjie Li,Chenxu Yu,Zhenyu Wang,Xinping Lin
Nine transformants of Rhodosporidium toruloides with significant changes in the carotenoid profile were obtained by Agrobacterium tumefaciens-mediated transformation, including a white, three red, and four yellow mutants. A red mutant A1-15-BRQ that showed a high torularhodin production was selected for culture condition optimization. Results indicated that the torularhodin yield was boosted with glucose as the carbon source, at a carbon/nitrogen ratio of 22, a loading volume of 75 mL, and 28 °C. The torularhodin yield of 21.3 mg/L consisting of 94.4% total carotenoids was obtained by Box–Behnken design experiments. The torularhodin yield was 17.0-fold higher than that of the wild type, with time shortened from 9 to 3 days. This study reports an effective strategy for improving torularhodin production and provides a candidate R. toruloides strain for highly selective production of torularhodin.
研究领域:
微生物学
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生物技术
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代谢工程
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DOI
PubMed
高影响因子
BIORESOURCE TECHNOLOGY
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2019
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IF 6.669
Xiaoyu Ma,Zhen Gao,Ming Gao,Chuanfu Wu,Qunhui Wang
This study aimed to clarify the composition changes of food waste after being placed for few days and propose a two-stage fermentation method to effectively convert food waste saccharified liquid (FWSL) into lipids by Rhodosporidium toruloides . Food waste generally needs 3–5 days to be transported and stored before treatment. The lactic acid concentration of FWSL produced from 5-days-placed-at-room-temperature food waste reached to 15 g/L. Lactic acid promoted yeast proliferation, and its main mechanism was the conversion of lactic acid into pyruvic acid, which could provide energy for yeast growth through TCA cycle. The optimal lipid concentration in the two-stage fermentation reached to 9.19 g/L, and lipid yield amounted to 0.204 g lipid/g total sugar; the values increased by 44.27% and 60.63%, respectively, when compared with those in traditional fermentation. This study could provide a strategy for food waste treatment closer to industrial production.
研究领域:
生物技术
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环境科学
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微生物学
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产品关联
DOI
PubMed
高影响因子
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