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Coexpression of fungal phytase and xylanase utilizing the cis-acting hydrolase element in Pichia pastoris
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Metadata
Document Title
Coexpression of fungal phytase and xylanase utilizing the cis-acting hydrolase element in Pichia pastoris
Author
Roongsawang N., Promdonkoy P., Wongwanichpokhin M., Sornlake W., Puseenam A., Eurwilaichitr L., Tanapongpipat S.
Name from Authors Collection
Scopus Author ID
55314049000
Scopus Author ID
26427756600
Scopus Author ID
6602764100
Affiliations
Microbial Cell Factory Laboratory, Bioresources Technology Unit, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Pathumthani, Thailand; Enzyme Technology Laboratory, Bioresources Technology Unit, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Pathumthani, Thailand
Type
Article
Source Title
FEMS Yeast Research
ISSN
15671356
Year
2010
Volume
10
Issue
7
Page
909-916
Open Access
All Open Access, Bronze
DOI
10.1111/j.1567-1364.2010.00669.x
Format
Abstract
Plant-based animal feed contains antinutritive agents, necessitating the addition of digestive enzymes in commercial feeds. Enzyme additives are costly because they are currently produced separately from recombinant sources. The coexpression of digestive enzymes in a single recombinant cell system would thus be advantageous. A coexpression system for the extracellular production of phytase and xylanase was established in Pichia pastoris yeast. The genes for each enzyme were fused in-frame with the α-factor secretion signal and linked by the 2A-peptide-encoding sequence. Each enzyme was expressed extracellularly as individual functional proteins. The specific activities of 2A-expressed phytase (PhyA-2A) and 2A-expressed xylanase (XylB-2A) were 9.3 and 97.3 U mg-1, respectively. Optimal PhyA-2A activity was observed at 55 °C and pH 5.0. PhyA-2A also exhibited broad pH stability from 2.5 to 7.0 and retained ~70% activity after heating at 90 °C for 5 min. Meanwhile, XylB-2A exhibited optimal activity at 50 °C and pH 5.5 and showed pH stability from 5.0 to 8.0. It retained >50% activity after incubation at 50 °C for 10 min. These enzyme properties are similar to those of individually expressed recombinant enzymes. In vitro digestibility test showed that PhyA-2A and XylB-2A are as efficient as individually expressed enzymes for hydrolyzing phytate and crude fiber in feedstuff, respectively. © 2010 Federation of European Microbiological Societies.
Keyword
2A peptide | cis-acting hydrolase element | Coexpression | Phytase | Xylanase
Industrial Classification
Knowledge Taxonomy Level 1
Knowledge Taxonomy Level 2
Knowledge Taxonomy Level 3
License
N/A
Rights
N/A
Publication Source
Scopus