一株希瓦氏菌植酸酶的特性分析及其在毕赤酵母中的表达。
A novel phytase from Citrobactergillenii: characterization and expression in Pichia pastoris (Komagataella pastoris).
机构信息
NRC «Kurchatov Institute» - GOSNIIGENETIKA, Kurchatov Genomic Center, NRC «Kurchatov Institute», 1-st Dorozhnyy proyezd, 1, Moscow, 117545, Russia.
NRC "Kurchatov Institute", Akademika Kurchatova pl., 1, Moscow, 123182, Russia.
出版信息
FEMS Microbiol Lett. 2021 Feb 4;368(2). doi: 10.1093/femsle/fnaa217.
The phyCg gene encoding a new phytase from Citrobacter gillenii was optimized, synthesized, cloned and expressed in Pichia pastoris. Analysis of the amino acid sequence of the enzyme showed that it belongs to the histidine acid phosphatase family. The amino acid sequence of the PhyCg phytase has the highest homology (73.49%) with a phytase sequence from Citrobacter braakii. The main characteristics for the purified recombinant phytase were established. The optimum pH and temperature were 4.5 and 50°C, respectively. The specific activity of the enzyme was 1577 U/mg. The Michaelis constant (Km) and the maximum reaction rate (Vmax) for sodium phytate were 0.185 mM and 2185 U/mg, respectively. The enzyme showed the pH and trypsin stability and had a high activity over a wide pH range.
从柠檬酸杆菌（Citrobacter gillenii）中优化、合成、克隆并表达了编码新植酸酶的 phyCg 基因，在毕赤酵母（Pichia pastoris）中表达。对该酶的氨基酸序列分析表明，它属于组氨酸酸性磷酸酶家族。PhyCg 植酸酶的氨基酸序列与柠檬酸杆菌（Citrobacter braakii）的植酸酶序列具有最高的同源性（73.49%）。纯化的重组植酸酶的主要特性得以确定。最佳 pH 值和温度分别为 4.5 和 50°C。该酶的比活为 1577 U/mg。植酸钠的米氏常数（Km）和最大反应速率（Vmax）分别为 0.185 mM 和 2185 U/mg。该酶表现出对 pH 值和胰蛋白酶的稳定性，在较宽的 pH 范围内具有较高的活性。
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