摘要
【目的】研究转录调控因子Bas1p和Bas2p协同作用对重组酿酒酵母(Saccharomyces cerevisiae)胞外c AMP产生的影响,初步优化发酵培养基。【方法】通过共整合表达策略,在c AMP产生菌酿酒酵母G5中超表达Bas1p和Bas2p,摇瓶发酵实验考察了Bas1p和Bas2p协同作用对菌株生长及胞外c AMP产生的影响,进一步考察了酵母粉和蛋白胨含量及前体物腺嘌呤对菌株生长和c AMP产生的影响。【结果】超表达Bas1p和Bas2p使菌株在1×YP培养基中发酵120 h时的c AMP产量较出发菌株提高51.4%,达到2 253.8μmol/L;将1×YP中的酵母粉和蛋白胨含量翻倍(即2×YP培养基)发酵120 h时的c AMP产量提高至4 450.4μmol/L;在2×YP培养基中添加0.5 g/L浓度的腺嘌呤时,c AMP产量进一步提高至5 314.3μmol/L。【结论】强化Bas1p和Bas2p的协同作用及相应地优化培养基组分有助于酿酒酵母胞外c AMP生产。
[Objective] The effect of cooperative interaction between Bas1 p and Bas2 p on c AMP production by recombinant Saccharomyces cerevisiae strain was studied, and fermentation medium was preliminarily optimized. [Methods] Bas1 p and Bas2 p were over-expressed by using the co-integrated expressing strategy in the c AMP-producing strain G5. The growth and extracellular c AMP production of the resultant strain were investigated by fermentation in shaking flasks. The effects of the amount of yeast powder and peptone, and the precursor adenine added in media were studied. [Results] The c AMP concentration at 120 h produced by the G5 strain over-expressing Bas1 p and Bas2 p was 2 253.8 μmol/L when fermentation medium 1×YP was used, which was increased by 51.4% co mpared with control strain. The product concentration was furt her increased to 4 450.4 μmol/L when the content of yeast powder and peptone in media was doubled. By adding 0.5 g/L adenine in media, the c AMP concentration at 120 h was further increased to 5 314.3 μmol/L. [Conclusion] All those results proved that the reinforced cooperative interaction between Bas1 p and Bas2 p as well as medium optimization can improve c AMP production.
出处
《微生物学通报》
CAS
CSCD
北大核心
2016年第2期370-378,共9页
Microbiology China
关键词
酿酒酵母
整合载体
环磷酸腺苷
BAS1-BAS2
腺嘌呤
Saccharomyces cerevisae
Integrating vector
Cyclic adenosine monophosphate
BAS1-BAS2
Adenine
