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Updated: Nov 24, 2025
Expression of Recombinant Proteins in the Methylotrophic Yeast Pichia pastoris
Published on: February 25, 2010
A novel phytase from Citrobactergillenii: characterization and expression in Pichia pastoris (Komagataella pastoris)
Artur A Tkachenko1, Anna N Kalinina2, Larisa N Borshchevskaya2
1NRC «Kurchatov Institute» - GOSNIIGENETIKA, Kurchatov Genomic Center, NRC «Kurchatov Institute», 1-st Dorozhnyy proyezd, 1, Moscow, 117545, Russia.
Abstract:
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.
