BAT基因改造对酿酒酵母高级醇生成量的影响

发布时间：2018-01-13 16:06

本文关键词：BAT基因改造对酿酒酵母高级醇生成量的影响　出处：《现代食品科技》2016年06期 　论文类型：期刊论文

【摘要】：酿酒酵母BAT基因编码支链氨基酸转氨酶,其中BAT1和BAT2基因分别编码线粒体和细胞质氨基酸转氨酶,位于细胞不同的位置导致二者的生理功能有所差异,BAT1基因在线粒体中倾向催化α-酮酸合成氨基酸,细胞质中的BAT2基因将氨基酸转化为α-酮酸,通过敲除BAT2以减少α-酮酸合成,过表达BAT1以增加α-酮酸消耗达到降低酿酒酵母高级醇的合成的目的。本研究以酿酒酵母AY15单倍体α5为出发菌株,结合融合PCR技术构建重组质粒p UC-BABPB1K,获得BA-PGK-BAT1-BB重组盒,并利用醋酸锂转化法和同源重组技术筛选出缺失BAT2基因同时过表达BAT1基因的突变株B-8,将其和亲本菌株α5、BAT2基因缺失菌株α5ΔBAT2进行酒精发酵实验,发酵结束后进行发酵性能和高级醇的测定。实验结果表明,与亲本菌株相比,异丁醇降低了25%,异戊醇降低了15%,活性戊醇降低了30%;与α5ΔBAT2菌株相比,异丁醇提高了0.5倍,异戊醇增加了0.1倍,活性戊醇增加了0.3倍。
[Abstract]:The Saccharomyces cerevisiae BAT gene encoding the branched chain amino acid aminotransferase, which BAT1 and BAT2 gene encoding the mitochondrial and cytoplasmic amino acid aminotransferase, located in cells lead to different physiological functions of the two different BAT1 gene in mitochondrial tendency in catalytic alpha keto acid amino acid synthesis, BAT2 gene in the cytoplasm of the conversion of amino acids to alpha keto acid. By knocking out the BAT2 to reduce the alpha keto acid synthesis, overexpression of BAT1 in order to increase the alpha keto acid to reduce the consumption of higher alcohol synthesis in Saccharomyces cerevisiae. In this study, AY15 of Saccharomyces cerevisiae haploid alpha 5 strain with recombinant plasmid P UC-BABPB1K fusion PCR, BA-PGK-BAT1-BB recombinant box, and homologous recombination technology screening of BAT2 gene deletion mutant B-8 and BAT1 gene expression by lithium acetate transformation method, the parent strain and alpha 5, alpha 5 Delta BAT2 gene deletion strain BAT 2 experiments of alcohol fermentation, determination of fermentation after fermentation performance and higher alcohols. The experimental results show that compared with the parent strain, isobutanol, isoamyl alcohol decreased by 25%, down 15%, active amyl alcohol decreased by 30%; compared with alpha 5 Delta BAT2 strains, isobutanol was 0.5 times higher than that of isoprene alcohol increased 0.1 times, active amyl alcohol increased 0.3 times.

【作者单位】：天津科技大学生物工程学院工业发酵微生物教育部重点实验室;
【基金】：天津市科委自然基金重点项目(14JCZDJC32900) 国家自然科学基金项目(31471724)
【分类号】：TS261.11
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