摘要
为了验证脱羧酶在S.cerevisiaeβ-苯乙醇合成途径中的调控作用,本文从S.cerevisiae S288C中克隆脱羧酶基因ARO10,并构建由3-磷酸甘油酸激酶基因PGK1组成型强启动子控制的ARO10基因表达载体pYES2-Ppgk-ARO10,将重组载体导入S.cerevisiae S288C,研究ARO10基因过量表达对重组菌株中β-苯乙醇合成的影响。经摇瓶实验测定,携带pYES2-Ppgk-ARO10的转化子SP10在发酵60h时β-苯乙醇产量达到最大量1.0g/L,较野生型的对照菌提高16.3%。研究结果表明,脱羧酶是S.cerevisiae S288C中β-苯乙醇生物合成途径的关键酶之一,增加ARO10基因表达量有利于提高β-苯乙醇产量,研究为构建β-苯乙醇高产工程菌株奠定了重要基础。
In order to verify the key role that the decarboxylase probably played in the synthesis pathway of β-phenethyl alcohol in S.cerevisiae,the decarboxylase gene ARO10 was cloned from S.cerevisiae S288C and then inserted into a constitutive expression vector controlled by the promoter of phosphoglyceate kinase gene PGK], generating a recombinant vector pYES2- Ppgk- ARO10. After pYES2- Ppgk- ARO10 was introduced into S. cerevisiae S288C by the lithium acetate transformation method, the influence of gene ARO10 over-expression on theβ-phenethyl alcohol production in the recombinant strain was evaluated.A maximum yield up to 1.0g/L of β-phenethyl alcohol was achieved when the recombinant strain SP10 was incubated for 60h in flask fermentation experiments,which was 16.3% higher than that of the wild strain.The results indicated that ARO10 over-expression significantly increased the β- phenethyl alcohol production in S. cerevisiae S288C, which demonstrated that the decarboxylase played a key role in the biosynthesis of β-phenethyl alcohol.This work established an important foundation for construction of the β-phenethyl alcohol high-producing genetically engineered strains.
出处
《食品工业科技》
CAS
CSCD
北大核心
2014年第3期155-159,共5页
Science and Technology of Food Industry
基金
北京市自然科学基金项目(5122008)
国家863计划项目(2012AA021502)
教育部科学技术研究重点项目(211101)
北京市教委KM201110011001)资助
关键词
Β-苯乙醇
酿酒酵母
脱羧酶
ARO10基因
过量表达
β- phenethyl alcohol
Saccharomyces cerevisiae
decarboxylase
gene AROIO
over- expression
