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首页> 外文期刊>African Journal of Biotechnology >Gene cloning of phenolic acid decarboxylase from Bacillus subtilis and expression in top-fermenting yeast strain
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Gene cloning of phenolic acid decarboxylase from Bacillus subtilis and expression in top-fermenting yeast strain

机译：枯草芽孢杆菌酚酸脱羧酶的基因克隆及其在发酵酵母菌株中的表达

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摘要

Phenolic acid decarboxylase (PADC) gene, encoding phenolic acid decarboxylase, was cloned from?Bacillus subtilis?and ligated with a shuttle vector YEp352 to generate a novel plasmid YPADC. By analysis of sequencing and the restriction endonuclease digestion, the validity of construction was proved. Subsequently, the new vector was successfully transformed into wild-type top-fermenting yeast strain W303-1A; the mutant yeast strain W303+padc?was obtained, which was tested on the laboratory-scale mashing and fermentation experiments. At the end of fermentation, the results showed an obvious increase of 4-vinylguaiacol content in top-fermented beers brewed with mutant yeasts. The final 4-vinylguaiacol concentration obtained with wild-type and mutant yeasts was 1.20 and 1.70mg/l, respectively. Additionally, the level of esters produced by the mutant strain was higher than that of the wild-type; there were therefore a marked clove-like and ester aroma in top-fermented beers brewed with the former. However, no evident differences were found in brewing characteristic between wild-type and mutant strains, especially the ability of utilizing fermentable sugar and reducing diacetyl. Taken together, these approaches indicated the possibility of cloning?PADC?gene and enhancing the concentration of 4-vinylguaiacol in top-fermented beers.
机译：从枯草芽孢杆菌中克隆出编码酚酸脱羧酶的苯酚酸脱羧酶（PADC）基因，并与穿梭载体YEp352连接，以产生新的质粒YPADC。通过测序分析和限制性内切酶消化，证明了构建的有效性。随后，新载体成功转化为野生型顶级发酵酵母菌株W303-1A;获得了突变酵母菌株W303 + padc ?，并在实验室规模的糖化和发酵实验中进行了测试。发酵结束时，结果显示，用突变酵母酿造的顶级发酵啤酒中4-乙烯基愈创木酚的含量明显增加。用野生型和突变型酵母获得的最终4-乙烯基愈创木酚的终浓度分别为1.20和1.70mg / l。另外，突变菌株产生的酯水平高于野生型。因此，用前者酿造的顶级发酵啤酒有明显的丁香和酯香气。但是，野生型和突变型菌株的酿造特性，特别是利用可发酵糖和还原二乙酰基的能力，没有发现明显的差异。综上所述，这些方法表明克隆顶部发酵啤酒中的PADCα基因和提高4-乙烯基愈创木酚浓度的可能性。

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