A phosphinothricin dehydrogenase mutant, a genetic engineering bacterium, and a one-pot multi-enzyme synchronous directed evolution method. The phosphinothricin dehydrogenase mutant is obtained by mutating the 164th amino acid from alanine to glycine, mutating the 205th arginine to lysine and mutating the 332nd threonine to alanine of phosphinothricin dehydrogenase derived from Pseudomonas fluorescens, and an amino acid sequence is as shown in SEQ ID No.1. The genetically engineered bacterium is obtained by introducing a gene of the phosphinothricin dehydrogenase mutant into a host cell. A coding gene of glucose dehydrogenase or a coding gene of formate dehydrogenase can also be introduced into the host cell to perform simultaneous directed evolution to overexpress the double genes. The one-pot multi-enzyme synchronous directed evolution method can obtain, by means of screening, a genetically engineered bacterium having greatly improved activity. Compared with catalytic processes such as transaminase, the L-PPT preparation method has a relatively simple process, a high conversion rate of raw materials, a conversion rate of up to 100%, and high stereo-selectivity.
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机译：磷酸丁蛋白脱氢酶突变体，遗传工程细菌和一种单盆多酶同步定向演化方法。通过将来自丙氨酸的第164个氨基酸突变为甘氨酸的第164个氨基酸来获得膦腈蛋白脱氢酶突变体，将第205天精氨酸突变并突变332nd苏氨酸至来自荧光荧光素的磷素脱氢酶的丙氨酸，并且氨基酸序列如SEQ ID NO所示.1。通过将膦酸脱氢酶突变体的基因引入宿主细胞来获得遗传工程细菌。还可以将葡萄糖脱氢酶的编码基因或甲酸脱氢酶的编码基因引入宿主细胞中，以进行同时定向的进化以过度表达双基因。通过筛选可以获得具有大大提高活性的基因工程细菌可以获得单盆多酶同步定向的进化方法。与催化方法如转氨酶等相比，L-PPT制备方法具有相对简单的方法，原料的高转化率，转化率高达100％，并且立体选择性高。
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