![](/images/graphics-bg.png)
Molecular Cloning and Optimization for High Level Expression of Cold-Adapted Serine Protease from Antarctic Yeast Glaciozyma antarctica PI12
Joint Authors
Basri, Mahiran
Raja Abd. Rahman, Raja Noor Zaliha
Alias, Norsyuhada
Ahmad Mazian, Mu’adz
Salleh, Abu Bakar
Source
Issue
Vol. 2014, Issue 2014 (31 Dec. 2014), pp.1-20, 20 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2014-06-30
Country of Publication
Egypt
No. of Pages
20
Main Subjects
Abstract EN
Psychrophilic basidiomycete yeast, Glaciozyma antarctica strain PI12, was shown to be a protease-producer.
Isolation of the PI12 protease gene from genomic and mRNA sequences allowed determination of 19 exons and 18 introns.
Full-length cDNA of PI12 protease gene was amplified by rapid amplification of cDNA ends (RACE) strategy with an open reading frame (ORF) of 2892 bp, coded for 963 amino acids.
PI12 protease showed low homology with the subtilisin-like protease from fungus Rhodosporidium toruloides (42% identity) and no homology to other psychrophilic proteases.
The gene encoding mature PI12 protease was cloned into Pichia pastoris expression vector, pPIC9, and positioned under the induction of methanol-alcohol oxidase (AOX) promoter.
The recombinant PI12 protease was efficiently secreted into the culture medium driven by the Saccharomyces cerevisiae α-factor signal sequence.
The highest protease production (28.3 U/ml) was obtained from P.
pastoris GS115 host (GpPro2) at 20°C after 72 hours of postinduction time with 0.5% (v/v) of methanol inducer.
The expressed protein was detected by SDS-PAGE and activity staining with a molecular weight of 99 kDa.
American Psychological Association (APA)
Alias, Norsyuhada& Ahmad Mazian, Mu’adz& Salleh, Abu Bakar& Basri, Mahiran& Raja Abd. Rahman, Raja Noor Zaliha. 2014. Molecular Cloning and Optimization for High Level Expression of Cold-Adapted Serine Protease from Antarctic Yeast Glaciozyma antarctica PI12. Enzyme Research،Vol. 2014, no. 2014, pp.1-20.
https://search.emarefa.net/detail/BIM-453837
Modern Language Association (MLA)
Alias, Norsyuhada…[et al.]. Molecular Cloning and Optimization for High Level Expression of Cold-Adapted Serine Protease from Antarctic Yeast Glaciozyma antarctica PI12. Enzyme Research No. 2014 (2014), pp.1-20.
https://search.emarefa.net/detail/BIM-453837
American Medical Association (AMA)
Alias, Norsyuhada& Ahmad Mazian, Mu’adz& Salleh, Abu Bakar& Basri, Mahiran& Raja Abd. Rahman, Raja Noor Zaliha. Molecular Cloning and Optimization for High Level Expression of Cold-Adapted Serine Protease from Antarctic Yeast Glaciozyma antarctica PI12. Enzyme Research. 2014. Vol. 2014, no. 2014, pp.1-20.
https://search.emarefa.net/detail/BIM-453837
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-453837