CRISPR/Cas9-Mediated Genome Editing of the Komagataella phaffii to Obtain a Phytase-Producer Markerless Strain.

Using CRISPR/Cas9 system, the recipient strains K. phaffii VKPM Y-5013 (His phenotype) and K. phaffii VKPM Y-5014 (Leu phenotype) were derived from the K.

Biochemical characterisation of glycosylated and deglycosylated forms of phytase from Cronobacter turicensis expressed in Pichia pastoris.

Phytases are valuable industrial enzymes widely used in animal feed production, and when expressed in yeast, they undergo glycosylation. Herein, heterologous expression in Pichia pastoris and biochemical characterisation of glycosylated and deglycosylated forms of a novel phytase from Cronobacter turicensis belonging to the histidine acid phosphatase family were successfully carried out. Mutants with deleted N-glycosylation sites (N136, N171, and N202) were constructed by site-directed mutagenesis and characterised.

A novel phytase from Citrobactergillenii: characterization and expression in Pichia pastoris (Komagataella pastoris).

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.