马克斯克鲁维酵母重组蛋白高产突变株的转录组分析  

Transcriptome Analysis of Recombinant Protein High-yielding Mutant Strain of Kluyveromyces marxianus

作　　者：吴若男 尹安琪 余垚[1,2] 周峻岗 吕红[1,2] WU Ruonan;YIN Anqi;YU Yao;ZHOU Jungang;L Hong(State Key Laboratory of Genetic Engineering,Fudan University Shanghai 200438,China;Engineering Research Center of Industrial Microorganisms,School of Life Sciences,Fudan University,Shanghai 200438,China)

机构地区：[1]复旦大学遗传工程国家重点实验室,上海200438 [2]复旦大学生命科学学院上海工业菌株工程技术研究中心,上海200438

出　　处：《复旦学报(自然科学版)》2025年第2期148-159,共12页Journal of Fudan University:Natural Science

基　　金：科技部重点研发计划项目(2021YFA0910601);上海科学技术委员会基地项目(2023ZX01)。

摘　　要：马克斯克鲁维酵母是一种生长速度快、蛋白质合成能力强的食品级酵母,具有研究价值与工业应用潜力。本研究以豆血红蛋白A(Leghemoglobin A,Lba)为靶蛋白,通过H 2O 2和UV联合诱变筛选获得马克斯克鲁维酵母重组蛋白高产突变株F6-30,其Lba产量提高67%。F6-30表现出高产通用性,表达其他异源蛋白如阿魏酸酯酶、甘露聚糖酶和木聚糖酶时产量分别提升111%、119%和39.4%。转录组测序分析显示在表达Lba的情况下F6-30与出发菌株相比在发酵24 h和60 h分别有1629个和1841个基因有显著差异,上调基因分别有604个和540个,下调基因分别有1025个和1301个。GO富集分析显示显著差异基因富集在翻译过程、RNA加工以及溶酶体和自噬相关GO条目;KEGG富集分析显示富集通路主要为核糖体生物发生、氨基酸合成代谢、核苷酸代谢等,表明突变株中蛋白质翻译途径显著加强,自噬途径及蛋白酶体途径以及氨基酸合成代谢被削弱。Kluyveromyces marxianus is a kind of food-grade yeast with fast growth rate and strong protein synthesis ability,which has research value and industrial application potential.In this study,leghemoglobin A(Lba)was used as the target protein,and the high-yielding mutant F6-30 was selected after the combined hydrogen peroxide and UV mutagenesis,and the Lba yield was increased by 67%.F6-30 showed high yield versatility,with the expression of other proteins such as AnfaeA Man330 and Xyn-CDBFV increasing the yield by 111%,119%and 39.4%,respectively.Transcriptome sequencing analysis showed that there were significant differences in 1629 and 1841 genes of F6-30 expressing leghemoglobin at 24 h and 60 h,respectively.Besides,604 and 540 genes were up-regulated and 1025 and 1301 genes were down-regulated respectively in 24 h and 60 h.GO enrichment analysis showed that significantly differentially expressed genes were enriched in the translation process,RNA processing,lysosome and autophagy,and KEGG enrichment analysis showed that significantly differentially expressed genes were enriched in ribosome biogenesis,amino acid anabolic metabolism,nucleotide metabolism,etc.,indicating that translation pathways were significantly enhanced,the autophagy and proteasome pathways and amino acid anabolism were weakened,and the respiratory pattern was biased towards anaerobic fermentation in the mutant strain.

关 键 词：马克斯克鲁维酵母 转录组 重组蛋白表达 豆血红蛋白A 

分 类 号：Q933[生物学—微生物学]