高糖胁迫对小球藻生长和类胡萝卜素合成的影响
Effects of high glucose stress on growth and carotenoids synthesis of Chlorella vulgaris
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摘要: 【目的】 减轻小球藻发酵过程中初始葡萄糖质量浓度对其生长及类胡萝卜素合成的抑制效应。【方法】 以生物量和类胡萝卜素产量为指标,采用紫外诱变联合葡萄糖质量浓度梯度胁迫定向驯化技术筛选耐高糖小球藻,从转录组水平分析高糖胁迫对小球藻生长的影响机制,并利用逆转录-定量聚合酶链反应(RT-qPCR)对分析结果进行验证。【结果】 定向驯化获得一株能耐受30 g/L葡萄糖质量浓度的小球藻菌株,其糖耐受性较普通小球藻提高了约50%,类胡萝卜素产量为5.38 mg/L;耐高糖小球藻的基因表达量FPKM值在区间分布上与对照组有明显差异,且二者表达模式的相似度较低;耐高糖小球藻中有4个与半胱氨酸/蛋氨酸代谢通路相关的上调差异表达基因(DEGs)使其细胞内的丙酸及其衍生物累积,进而抑制其生长,其中gene2.1.1.14、gene2.5.1.6和gene3.5.99.7的表达相较于对照组分别上调184.74倍、175.68倍和179.28倍。【结论】 紫外诱变联合梯度葡萄糖质量浓度胁迫的定向驯化策略可有效提升小球藻的糖耐受性,而半胱氨酸/蛋氨酸代谢通路的异常调控是高糖环境抑制小球藻生长的重要分子机制之一。Abstract: 【Objective】 This study aimed to alleviate the inhibitory effect of high initial glucose concentration on the growth and carotenoid synthesis of Chlorella vulgaris during fermentation. 【Methods】 High glucose-tolerant C.vulgaris strains were screened with biomass and carotenoid yield as indicators, using UV mutagenesis combined with directional domestication under glucose concentration gradient stress. The mechanism underlying the effect of high glucose stress on the growth of C.vulgaris was analyzed at the transcriptome level, and the result were verified by RT-qPCR. 【Results】 A C.vulgaris strain capable of tolerating a glucose concentration of 30 g/L was obtained through directional domestication. Compared with the original strain, its glucose tolerance increased by approximately 50%, and the carotenoid yield reached 5.38 mg/L. The FPKM density distribution of gene expression in the high glucose-tolerant strain differed significantly from that of the control group, and the two expression patterns showed low similarity. Four upregulated differentially expressed genes (DEGs) related to the cysteine and methionine metabolism pathway in the high glucose-tolerant strain led to the accumulation of propionate and its derivatives in cells, thereby inhibiting growth. The expression levels of gene 2.1.1.14, gene 2.5.1.6, and gene 3.5.99.7 were upregulated 184.74-fold, 175.68-fold, and 179.28-fold, respectively, compared with the control group. 【Conclusion】 The strategy of UV mutagenesis combined with directional domestication under glucose concentration gradient stress can effectively improve the glucose tolerance of C.vulgaris, and the abnormal regulation of the cysteine/methionine metabolism pathway is one of the important molecular mechanisms by which high glucose stress inhibits its growth.
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[1]
郑力铭,王鹏超.微生物合成四萜类化合物研究进展[J].中国生物工程杂志,2023,43(10):96-108.
ZHENG L M,WANG P C.Progress in the synthesis of tetraterpenes by microorganisms[J].China Biotechnology,2023,43(10):96-108. -
[2]
张志平,段乃心,魏湘楠,等.基于光胁迫粘红酵母合成高值化类胡萝卜素发酵条件优化[J].轻工学报,2022,37(4):10-17.
ZHANG Z P,DUAN N X,WEI X N,et al.Optimization of fermentation conditions for high value carotenoid synthesis by Rhodotorula glutinis under light stress[J].Journal of Light Industry,2022,37(4):10-17. -
[3]
周琳,梁轩铭,赵磊.天然类胡萝卜素的生物合成研究进展[J].生物技术通报,2022,38(7):119-127.
ZHOU L,LIANG X M,ZHAO L.Biosynthesis of natural carotenoids:Progress and perspective[J].Biotechnology Bulletin,2022,38(7):119-127. -
[4]
MARTÍNEZ-CÁMARA S,IBAÑEZ A,RUBIO S,et al.Main carotenoids produced by microorganisms[J].Encyclopedia,2021,1(4):1223-1245.
-
[5]
LIU Z Y,YU W L,NOMURA C T,et al.Increased flux through the TCA cycle enhances bacitracin production by Bacillus licheniformis DW2[J].Applied Microbiology and Biotechnology,2018,102(16):6935-6946.
-
[6]
HAN J Y,SONG J M,SEO S H,et al.Ty1-fused protein-body formation for spatial organization of metabolic pathways in Saccharomyces cerevisiae[J].Biotechnology and Bioengineering,2018,115(3):694-704.
-
[7]
BIAN Q,JIAO X,CHEN Y,et al.Hierarchical dynamic regulation of Saccharomyces cerevisiae for enhanced lutein biosynthesis[J].Biotechnology and Bioengineering,2023,120(2):536-552.
-
[8]
刘晓桐,孙欣宇,刘玥,等.诱变筛选高产虾青素法夫酵母及发酵条件优化[J].大连工业大学学报,2021,40(6):396-400.
LIU X T,SUN X Y,LIU Y,et al.Screening of high astaxanthin producing Phaffia rhodozyma by mutagenesis and optimization of fermentation conditions[J].Journal of Dalian Polytechnic University,2021,40(6):396-400. -
[9]
VILLEGAS-MÉNDEZ M Á,MONTAÑEZ J,CONTRERAS-ESQUIVEL J C,et al.Scale-up and fed-batch cultivation strategy for the enhanced co-production of microbial lipids and carotenoids using renewable waste feedstock[J].Journal of Environmental Management,2023,339:117866.
-
[10]
JIN D,ZHANG X N,ZHOU L,et al.Emerging applications and mechanisms of algal-bacterial symbiosis on sustainable wastewater treatment:A comprehensive review[J].Journal of Water Process Engineering,2024,65:105912.
-
[11]
GONG G P,WU B,LIU L P,et al.Enhanced biomass and lipid production by light exposure with mixed culture of Rhodotorula glutinis and Chlorella vulgaris using acetate as sole carbon source[J].Bioresource Technology,2022,364:128139.
-
[12]
刘方舟,任洪艳,陈红芬,等.不同碳源对普通小球藻和粘红酵母共培养产油脂的影响[J].基因组学与应用生物学,2020,39(8):3612-3619.
LIU F Z,REN H Y,CHEN H F,et al.Effects of different carbon sources on lipid production in co-culture of Chlorella vulgaris and Rhodotorula glutinis[J].Genomics and Applied Biology,2020,39(8):3612-3619. -
[13]
杜梦帆,王露.不同碳源对小球藻生长的影响[J].生物化工,2022,8(4):93-95.
DU M F,WANG L.Effects of different carbon sources on the growth of Chlorella[J].Biological Chemical Engineering,2022,8(4):93-95. -
[14]
FERRO L,GOJKOVIC Z,MUÑOZ R,et al.Growth performance and nutrient removal of a Chlorella vulgaris-Rhizobium sp.co-culture during mixotrophic feed-batch cultivation in synthetic wastewater[J].Algal Research,2019,44:101690.
-
[15]
朱丽花,马延琴,纪彦宇,等.产类胡萝卜素酵母菌的筛选及色素稳定性分析[J].中国酿造,2021,40(9):139-144.
ZHU L H,MA Y Q,JI Y Y,et al.Screening of carotenoid-producing yeast and pigment stability analysis[J].China Brewing,2021,40(9):139-144. -
[16]
汤冬,张国森,赵晓芳.基于二代测序技术的转录组测序生物信息分析[J].河南大学学报(医学版),2019,38(1):67-76.
TANG D,ZHANG G S,ZHAO X F.Bioinformatics analysis of transcriptome sequencing based on next generation sequencing[J].Journal of Henan University (Medical Science),2019,38(1):67-76. -
[17]
ZHANG Z P,PANG Z J,XU S Y,et al.Improved carotenoid productivity and COD removal efficiency by co-culture of Rhodotorula glutinis and Chlorella vulgaris using starch wastewaters as raw material[J].Applied Biochemistry and Biotechnology,2019,189(1):193-205.
-
[18]
梁长利,郑训艺,郑祖凤,等.有机碳源对异养小球藻生长、蛋白质和叶绿素含量的影响[J].广东化工,2021,48(13):71-73.
LIANG C L,ZHENG X Y,ZHENG Z F,et al.Influence of organic carbon sources on the growth,protein and chlorophyll content of heterotrophic Chlorella vulgaris[J].Guangdong Chemical Industry,2021,48(13):71-73. -
[19]
JOUN J,SIROHI R,SIM S J.The effects of acetate and glucose on carbon fixation and carbon utilization in mixotrophy of Haematococcus Pluvialis[J].Bioresource Technology,2023,367:128218.
-
[20]
CAI Y H,LIU Y H,LIU T Y,et al.Heterotrophic cultivation of Chlorella vulgaris using broken rice hydrolysate as carbon source for biomass and pigment production[J].Bioresource Technology,2021,323:124607.
-
[21]
郎筱宇,刘志媛,徐梦,等.葡萄糖对小球藻(Chlorella sp.HN08)光合作用和生长的影响[J].微生物学报,2017,57(4):550-559.
LANG X Y,LIU Z Y,XU M,et al.Effects of glucose on photosynthesis and growth of Chlorella sp.HN08 cells[J].Acta Microbiologica Sinica,2017,57(4):550-559.
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