A novel phytase from Citrobacter gillenii: characterization and expression in Pichia pastoris (Komagataella pastoris)

被引：4
|
作者：
Tkachenko, Artur A. [1 ]
Kalinina, Anna N. [2 ]
Borshchevskaya, Larisa N. [2 ]
Sineoky, Sergey P. [2 ]
Gordeeva, Tatiana L. [1 ,2 ]
机构：
[1] NRC Kurchatov Inst, NRC Kurchatov Inst GOSNIIGENETIKA, Kurchatov Genom Ctr, 1 St Dorozhnyy Proyezd,1, Moscow 117545, Russia
[2] NRC Kurchatov Inst, Akad Kurchatova Pl 1, Moscow 123182, Russia
来源：
FEMS MICROBIOLOGY LETTERS | 2021年 / 368卷 / 02期
关键词：
phytate; phytase; Citrobacter gillenii; Pichia pastoris; protein expression; characterization; ESCHERICHIA-COLI PHYTASE; HIGH-LEVEL EXPRESSION; HETEROLOGOUS EXPRESSION; CODON OPTIMIZATION; MICROBIAL PHYTASE; STABLE PHYTASE; GENE; APPA; OVEREXPRESSION; FREUNDII;
D O I：
10.1093/femsle/fnaa217
中图分类号：
Q93 [微生物学];
学科分类号：
071005 ; 100705 ;
摘要：
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 degrees C, respectively. The specific activity of the enzyme was 1577 U/mg. The Michaelis constant (K-m) and the maximum reaction rate (V-max) 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.
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页数：8
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