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中国精品科技期刊2020
李童, 孙军勇, 吴殿辉, 李晓敏, 谢广发, 陆健. YDL080C和LEU2基因敲除对工业黄酒酵母异戊醇生成量的影响[J]. 食品工业科技, 2015, (15): 189-193. DOI: 10.13386/j.issn1002-0306.2015.15.032
引用本文: 李童, 孙军勇, 吴殿辉, 李晓敏, 谢广发, 陆健. YDL080C和LEU2基因敲除对工业黄酒酵母异戊醇生成量的影响[J]. 食品工业科技, 2015, (15): 189-193. DOI: 10.13386/j.issn1002-0306.2015.15.032
LI Tong, SUN Jun-yong, WU Dian-hui, LI Xiao-min, XIE Guang-fa, LU Jian. Effect of YDL080C and LEU2 gene knockout on isoamyl alcohol production in industrial yellow rice wine yeast[J]. Science and Technology of Food Industry, 2015, (15): 189-193. DOI: 10.13386/j.issn1002-0306.2015.15.032
Citation: LI Tong, SUN Jun-yong, WU Dian-hui, LI Xiao-min, XIE Guang-fa, LU Jian. Effect of YDL080C and LEU2 gene knockout on isoamyl alcohol production in industrial yellow rice wine yeast[J]. Science and Technology of Food Industry, 2015, (15): 189-193. DOI: 10.13386/j.issn1002-0306.2015.15.032

YDL080C和LEU2基因敲除对工业黄酒酵母异戊醇生成量的影响

Effect of YDL080C and LEU2 gene knockout on isoamyl alcohol production in industrial yellow rice wine yeast

  • 摘要: 通过敲除生成途径中关键酶的编码基因,对异戊醇在工业黄酒酵母N85中的生成进行了研究。采用融合PCR技术,分别构建类丙酮酸脱羧酶基因(YDL080C)缺失组件"YDL080C-URA3-YDL080C"和β-异丙基苹果酸脱氢酶基因(LEU2)缺失组件"LEU2-URA3-LEU2",转化工业黄酒酵母N85尿嘧啶缺陷型单倍体Na-u(MATa ura3),获得单倍体工程菌Na-y和Na-l。将转化子与亲本分别进行实验室规模的黄酒发酵实验,结果 YDL080C缺失工程菌的异戊醇含量与亲本相比没有明显变化,说明类丙酮酸脱羧酶不是该途径关键酶;LEU2缺失工程菌的异戊醇含量降低16.16%,说明糖代谢途径存在,但只占小部分,而其他理化指标无明显差异。 

     

    Abstract: The production of isoamyl alcohol,was studied in industrial rice wine yeast N85 with the method of gene knockout.Disruption cassette of “ YDL080C- URA3- YDL080 C ”and “ LEU2- URA3- LEU2 ”were constructed using fusion PCR,and then electro- transformed into the Δura3 haploid yeast strain( MATa ura3),respectively.Finally the engineered haploids Na- y and Na- l were obtained. Lab- scale fermentation of yellow rice wine was preceded with Na- y and Na- l,respectively. The results showed that the concentration of isoamyl alcohol was almost invariable between the engineered strain Na- y and the parental haploid. On the contrary,the content of isoamyl alcohol was reduced by 16.16% with the engineered strain Na- l.There was no difference in physical and chemical indexes between the engineered strain and parental strain.

     

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