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首页> 外文期刊>BMC Biotechnology >Optimization of glutathione production in batch and fed-batch cultures by the wild-type and recombinant strains of the methylotrophic yeast Hansenula polymorpha DL-1
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Optimization of glutathione production in batch and fed-batch cultures by the wild-type and recombinant strains of the methylotrophic yeast Hansenula polymorpha DL-1

机译：甲基营养酵母多形汉逊酵母DL-1的野生型和重组菌株优化分批和补料分批培养中谷胱甘肽的生产

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Background Tripeptide glutathione (gamma-glutamyl-L-cysteinyl-glycine) is the most abundant non-protein thiol that protects cells from metabolic and oxidative stresses and is widely used as medicine, food additives and in cosmetic industry. The methylotrophic yeast Hansenula polymorpha is regarded as a rich source of glutathione due to the role of this thiol in detoxifications of key intermediates of methanol metabolism. Cellular and extracellular glutathione production of H. polymorpha DL-1 in the wild type and recombinant strains which overexpress genes of glutathione biosynthesis ( GSH2 ) and its precursor cysteine ( MET4 ) was studied. Results Glutathione producing capacity of H. polymorpha DL-1 depending on parameters of cultivation (dissolved oxygen tension, pH, stirrer speed), carbon substrate (glucose, methanol) and type of overexpressed genes of glutathione and its precursor biosynthesis during batch and fed-batch fermentations were studied. Under optimized conditions of glucose fed-batch cultivation, the glutathione productivity of the engineered strains was increased from ~900 up to ~ 2300 mg of Total Intracellular Glutathione (TIG) or GSH+GSSGin, per liter of culture medium. Meantime, methanol fed-batch cultivation of one of the recombinant strains allowed achieving the extracellular glutathione productivity up to 250 mg of Total Extracellular Glutathione (TEG) or GSH+GSSGex, per liter of the culture medium. Conclusions H. polymorpha is an competitive glutathione producer as compared to other known yeast and bacteria strains ( Saccharomyces cerevisiae, Candida utilis, Escherichia coli, Lactococcus lactis etc.) with good perspectives for further improvement especially for production of extracellular form of glutathione.
机译：背景技术三肽谷胱甘肽（γ-谷氨酰基-L-半胱氨酸-甘氨酸）是最丰富的非蛋白质硫醇，可保护细胞免受代谢和氧化应激，广泛用作药物，食品添加剂和化妆品工业。甲基营养型多形汉逊酵母被认为是谷胱甘肽的丰富来源，因为该硫醇在甲醇代谢关键中间体的解毒中发挥了作用。研究了野生型多形汉逊酵母DL-1的细胞和细胞外谷胱甘肽的产生以及过量表达谷胱甘肽生物合成基因（GSH2）及其前体半胱氨酸（MET4）的重组菌株。结果多形汉逊酵母DL-1的谷胱甘肽生产能力取决于培养参数（溶解氧张力，pH，搅拌器速度），碳底物（葡萄糖，甲醇）以及谷胱甘肽过表达基因的类型及其在分批和进料期间的前体生物合成。研究了分批发酵。在分批补料葡萄糖培养的最佳条件下，工程菌株的谷胱甘肽生产率从约900 mg增至〜2300 mg总细胞内谷胱甘肽（TIG）或GSH + GSSG 每升培养基中同时，对其中一种重组菌株进行甲醇补料分批培养，可实现高达250 mg的总细胞外谷胱甘肽（TEG）或GSH + GSSG ex ，每升培养基。结论与其他已知的酵母和细菌菌株（酿酒酵母，假丝酵母​​，大肠杆菌，乳酸乳球菌等）相比，多形汉逊酵母是竞争性的谷胱甘肽生产者，具有进一步改进的良好前景，尤其是用于生产细胞外形式的谷胱甘肽。

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