Development of a <i>Pichia pastoris </i> strain for secretory production of recombinant alkaline phytase

Thi Hoang Lan Vo; Quoc Cuong Chau; Van Hau Nguyen; Hieu Nghia Nguyen

doi:10.15625/vjbt-21940

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Authors

Thi Hoang Lan Vo $^{1}$ Department of Molecular and Environmental Biotechnology, Faculty of Biology and Biotechnology, VNU-HCM University of Science, 227 Nguyen Van Cu street, Ward 4, District 5, Ho Chi Minh city, Vietnam
$^{2}$ Vietnam National University Ho Chi Minh City, Linh Trung Ward, Thu Duc city, Ho Chi Minh city, Vietnam https://orcid.org/0009-0005-3461-8954
Quoc Cuong Chau $^{1}$ Department of Molecular and Environmental Biotechnology, Faculty of Biology and Biotechnology, VNU-HCM University of Science, 227 Nguyen Van Cu street, Ward 4, District 5, Ho Chi Minh city, Vietnam
$^{2}$ Vietnam National University Ho Chi Minh City, Linh Trung Ward, Thu Duc city, Ho Chi Minh city, Vietnam https://orcid.org/0009-0008-8194-1318
Van Hau Nguyen $^{1}$ Department of Molecular and Environmental Biotechnology, Faculty of Biology and Biotechnology, VNU-HCM University of Science, 227 Nguyen Van Cu street, Ward 4, District 5, Ho Chi Minh city, Vietnam
$^{2}$ Vietnam National University Ho Chi Minh City, Linh Trung Ward, Thu Duc city, Ho Chi Minh city, Vietnam https://orcid.org/0000-0002-3939-4348
Hieu Nghia Nguyen $^{1}$ Department of Molecular and Environmental Biotechnology, Faculty of Biology and Biotechnology, VNU-HCM University of Science, 227 Nguyen Van Cu street, Ward 4, District 5, Ho Chi Minh city, Vietnam
$^{2}$ Vietnam National University Ho Chi Minh City, Linh Trung Ward, Thu Duc city, Ho Chi Minh city, Vietnam https://orcid.org/0000-0003-3151-2781

DOI:

https://doi.org/10.15625/vjbt-21940

Keywords:

alkaline phytase, aquaculture feed enzyme, Pichia pastoris, phytate hydrolysis, secretion expression

Abstract

Recombinant phytase produced by the Pichia pastoris expression system is widely utilized in the food and animal feed industries. Alkaline phytases are preferred as aquafeed supplements due to their efficiency in degrading antinutritional phytates. In this study, we engineered a recombinant P. pastoris strain to secrete alkaline phytase derived from Bacillus subtilis, with the aim of further application in the aquafeed industry. The recombinant vector, pPICZαA-phy, carrying the phytase-encoding gene (phy), was successfully constructed and integrated into the genome of P. pastoris X33. The resulting recombinant strain, P. pastoris X33::phy, exhibited a methanol-utilization positive phenotype (Mut⁺) and utilized methanol as a carbon source. Secretion of the recombinant alkaline phytase by P. pastoris X33::phy was confirmed through western blot analysis. Enzymatic activity assay demonstrated that this engineered yeast strain is capable of secreting alkaline phytase, which was able to hydrolyze phytate under mildly alkaline conditions (pH 7.5). These results suggest the potential of this strain for industrial applications, particularly in aquafeed production.

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Development of a Pichia pastoris strain for secretory production of recombinant alkaline phytase

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