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首页> 外文会议>International conference on applied biotechnology >Effect of Proteinase A Propeptide Deletion on its Enzyme Activity in Saccharomyces cerevisiae
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Effect of Proteinase A Propeptide Deletion on its Enzyme Activity in Saccharomyces cerevisiae

机译：酿酒酵母中蛋白酶A前肽的缺失对其酶活性的影响

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Two expression plasmids YCP-EP and YCP-54A respectively expressing the complete PEP4 gene and the incomplete PEP4 gene without the prosequence (encode the propeptide of proteinase A) from Saccharomyces cerevisiae W303-1A were constructed and transferred into S. cerevisiae W303-K22 (ΔPEP4). The yeast transformants WYCP-EP and WYCP-54A were obtained. Meanwhile, a mutant strain W54A deleted the prosequence of PEP4 was obtained by homologous recombination. The proteinase A enzyme activities of the yeast transformants WYCP-EP, WYCP-54A and the mutant strain W54A were detected by fluorescent substrate, respectively. The results showed that only the strain WYCP-EP was able to produce the active proteinase A. Consequently, the results confirmed that the propeptide was required for formation of active mature proteinase A, and it provided the theoretical basis for the construction of genetic engineering strain with low-proteinase A by the deletion of prosequence.
机译：构建了两个表达质粒YCP-EP和YCP-54A，它们分别表达酿酒酵母W303-1A的不带有序列（编码蛋白酶A的前肽）的完整PEP4基因和不完整PEP4基因，并转移到酿酒酵母W303-K22中（ ΔPEP4）。获得酵母转化体WYCP-EP和WYCP-54A。同时，通过同源重组获得了缺失PEP4序列的突变株W54A。通过荧光底物分别检测酵母转化体WYCP-EP，WYCP-54A和突变株W54A的蛋白酶A酶活性。结果表明，只有菌株WYCP-EP能够产生活性蛋白酶A。因此，结果证实前肽是形成活性成熟蛋白酶A所必需的，这为基因工程菌株的构建提供了理论依据。与低蛋白酶A有关的前序缺失。

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