多形汉逊酵母提高生长性能的培养基优化  

Medium optimization for growth of Ogataea polymorpha

作　　者：刘爽[1,2] 高教琪[1] 薛闯 周雍进[1] LIU Shuang;GAO Jiaoqi;XUE Chuang;ZHOU Yongjin(Dalian Institute of Chemical Physics,Chinese Academy of Sciences,Dalian 116031,China;School of Bioengineering,Dalian University of Technology,Dalian 116024,China)

机构地区：[1]中国科学院大连化学物理研究所,辽宁大连116031 [2]大连理工大学生物工程学院,辽宁大连116024

出　　处：《生物加工过程》2020年第1期116-125,共10页Chinese Journal of Bioprocess Engineering

基　　金：国家自然科学基金青年项目(21808216);中国科学院大连化学物理研究所创新基金(DICP I201920);大连市创新基金(2019J12GX030)

摘　　要：为了提高甲醇酵母多形汉逊的生长性能并降低发酵成本,对其培养基中维生素和氨基酸有效成分进行优化。通过单因素实验和响应面实验相结合,最终确定了两种优化后的培养基组成,分别是以葡萄糖为碳源时的培养基,(NH4)2 SO42.5 g/L、KH2 PO414.4 g/L、MgSO4·7H2O 0.5 g/L、trace-metal 2 mL/L、亮氨酸60 mg/L、葡萄糖20 g/L、生物素0.005 mg/L、甲硫氨酸40 mg/L、谷氨酸200 mg/L、天冬氨酸200 mg/L;以甲醇为碳源时的培养基,(NH4)2 SO42.5 g/L、KH2 PO414.4 g/L、MgSO4·7H2O 0.5 g/L、trace-metal 2 mL/L、亮氨酸60 mg/L、甲醇10 g/L、生物素0.005 mg/L、维生素B11.04 mg/L。这两种培养基分别将菌种生物量(OD 600)提高了6倍和1.5倍。优化后的葡萄糖培养基细胞中丙酮酸羧化酶活性提高了1倍,甲醇培养基细胞中丙酮酸脱氢酶酶活性提高3倍以上,丙酮酸羧化酶活性提高了8倍左右。综上,本研究为汉逊酵母大规模发酵提供了参考。In order to improve Ogataea polymorpha cell growth and reduce the fermentation costs,vitamin and amino acid components of culture medium were optimized.Through the combination of single factor and response surface experiments,two optimized media were finally determined in glucose and methanol-based culture,respectively.When using glucose as the sole carbon source,the optimal medium was composed of(NH4)2 SO4 2.5 g/L,KH2 PO4 14.4 g/L,MgSO4·7H2O 0.5 g/L,trace-metal 2 mL/L,leucine 60 mg/L,glucose 20 g/L,biotin 0.005 mg/L,methionine 40 mg/L,glutamic acid 200 mg/L,and aspartic acid 200 mg/L.When using methanol as the sole carbon source,the components of optimized culture were:(NH4)2SO4 2.5 g/L,KH2PO4 14.4 g/L,MgSO4·7H2O 0.5 g/L,trace-metal 2 mL/L,leucine 60 mg/L,methanol 10 g/L,biotin 0.005 mg/L and thiamin 1.04 mg/L.These two media improved the biomass yields(OD 600)by 6 times and 1.5 times,respectively.Optimized glucose-basea medium increased mraceuarar activity of pyrurate carboxylase by over 1 fold,and the activities of pyruvate dehydrogenase and pyruvate carboxylase were improved by 3-,and 8-fold,respectively,in methand medium.In a word,this study may lay a good foundation for industrial-scode cultivation of O.polymorpha cell.

关 键 词：多形汉逊酵母 细胞生长 维生素 氨基酸 培养基优化 基础盐培养基 甲醇 

分 类 号：Q815[生物学—生物工程]