分泌糖化酶的工業啤酒酵母菌的構建

Construction of Brewing Yeast Secreting Glucoamylase

唐國敏(Guo-Min Tang)

《Fungal Science》 12卷1&2期 (1997/06) Pp. 43-49

https://doi.org/10.7099/FS.199706.0043

摘要

以黑麯黴糖化酶高產株Aspergillus niger T21為基因源合成糖化酶cDNA，構建了由酵母磷酸甘油激酶(PGK)基因啓動子和終止子引導表達，糖化酶自身信號肽序列引導分泌，由酵母Ty因數的δ序列引導同源整合的整合型糖化酶表達分泌質粒pKG1。採用pKG1與G418抗性質粒的共轉化，將糖化醃cDNA引入啤酒酵母B48並整合到細胞染色體DNA上，獲得了穩定的轉化子。搖瓶發酵試驗表明，引入糖化酶基因可將啤酒發酵度從69%提高到80%，但並不影響菌株生長和啤酒風味。

關鍵字

啤酒酵母 ； 表達 ； 糖化醣cDNA ； 整合 ； 分泌

並列摘要

Glucoamylase (GA) cDNA was synthesized from a GA overproducing Aspergillus niger strain T21. The integrative type plasmid pKG1 for expression and secretion of glusoamylase was constructed by insertion of GA cDNA between yeast phosphoglycerate kinase promoter and terminator regions. The plasmid contained the δ sequence of yeast Ty element, which was used as a homologous fragment for integration. The GA cDNA was introduced into a brewing yeast B48 and integrated onto its chromosomal DNA by cotransfromation of pKG1 and a YEP type plasmid pBEJ16 carrying G418 resistance. The stable transformants were selected and used for flask fermentation test. The results of test showed that the fermentation rate was raised from original 69% to 80%. The growth properties of the transformants and the beer taste were judged to be good.

並列關鍵字

Brewing yeast ； expression ； glucoamylase cDNA ； integration ； secretion