Abstract

A DNA sequence from Kosakonia sacchari that, according to automated computer analysis, is believed to correspond to the gene for histidine-acid phytase has been selected from the GenBank database. The sequence was optimized for codon composition, synthesized, cloned, and expressed in Pichia pastoris. The main characteristics of the purified recombinant enzyme were determined. It was established that a pH of 4.5 and a temperature of 50°C are optimal for phytase functioning. The specific activity, Michaelis constant (K_m), and maximum reaction rate (V_max) with phytate as a substrate were 1470 U/mg, 193 μM and 2167 μmol/(min mg), respectively. It was shown that the enzyme was characterized by a wide range of working pH levels. Therefore, due to its properties, this new recombinant phytase can be considered a high-potential enzyme for agrobiotechnology.

中文翻译：

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糖酵母（Kosakonia sacchari）的新型重组植酸酶：特征及其生物技术潜力。

摘要

从GenBank数据库中选择了根据自动计算机分析，被认为对应于组氨酸-酸植酸酶基因的来自Kosakonia sacchar i的DNA序列。该序列针对密码子组成进行了优化，合成，克隆并在巴斯德毕赤酵母中表达。确定了纯化的重组酶的主要特征。确定植酸酶功能的最佳pH值为4.5，温度为50°C。比活，米氏常数（K _m）和最大反应速率（V _max）以肌醇六磷酸为底物的）分别为1470 U / mg，193μM和2167μmol/（min mg）。结果表明，该酶具有广泛的工作pH值。因此，由于其特性，这种新的重组植酸酶可以被认为是农业生物技术的一种高潜力酶。