CAJ | 학술논문

LEU1基因编码异丙基苹果酸合成酶，为了研究该基因对酿酒酵母高级醇生成量的影响，以酿酒酵母AY15单倍体A8为出发菌株，通过构建重组质粒pUC-LABK获得LA-KanMX-LB重组盒，并利用醋酸锂转化法和同源重组技术，筛选出LEU1基因缺失的突变株A-L9。将突变株和亲本菌株分别进行酒精发酵实验，发酵结束后进行发酵性能和高级醇生成量的测定。两种发酵条件下的实验结果表明，与亲本菌株相比，突变株的正丙醇生成量分别提高了0.18倍和0.47倍，异丁醇的生成量分别提高了0.52倍和1.58倍，而异戊醇的生成量则分别降低了0.29倍和0.51倍。
LEU1기인편마이병기평과산합성매，위료연구해기인대양주효모고급순생성량적영향，이양주효모AY15단배체A8위출발균주，통과구건중조질립pUC-LABK획득LA-KanMX-LB중조합，병이용작산리전화법화동원중조기술，사선출LEU1기인결실적돌변주A-L9。장돌변주화친본균주분별진행주정발효실험，발효결속후진행발효성능화고급순생성량적측정。량충발효조건하적실험결과표명，여친본균주상비，돌변주적정병순생성량분별제고료0.18배화0.47배，이정순적생성량분별제고료0.52배화1.58배，이이무순적생성량칙분별강저료0.29배화0.51배。
In order to study the effects of LEU1 gene (encoding isopropylmalate isomerase) deletion on higher alcohols yield, the recombinant plasmid pUC-LABK was constructed and the recombinant cassette LA-KanMX-LB was gained by PCR amplification. Then LEU1 gene defi-cient mutant A-L9 was constructed from S.cerevisiae haploid A8 by yeast transformation and homologous recombination. Then fermentation performance and higher alcohols yield of parental haploid and LEU1 mutant were determined at the end of ethanol fermentation. The results of two different kinds of culture conditions showed that, the yield of n-propanol in A-L9 were 0.18 and 0.47 fold higher than that of parental hap-loid respectively, and the yield of isobutanol increased by 0.52 and 1.58 fold, compared with parental haploid. However, the content of isoamyl alcohol in A-L9 reduced by 29.19%and 51.03%.