High-Level Production of Recombinant Chicken Cystatin by Pichia pastoris and Its Application in Mackerel Surimi

A high level of the secreted form of recombinant chicken cystatin was expressed in Pichia pastoris X-33 by chromosomal integration of multiple copies of an expression cassette containing chicken cystatin under the control of glyceraldehyde-3-phosphate dehydrogenase promoter. The inhibition ability o...

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Bibliographic Details
Published in:Journal of agricultural and food chemistry 2001-02, Vol.49 (2), p.641-646
Main Authors: Chen, Gen-Hung, Tang, Shye-Jye, Chen, Ching-San, Jiang, Shann-Tzong
Format: Article
Language:English
Subjects:
Animals
Base Sequence
Biological and medical sciences
Chickens
Cloning, Molecular - methods
cystatin
Cystatins - biosynthesis
Cystatins - genetics
Cystatins - metabolism
DNA Primers
Endopeptidases - metabolism
Fish and seafood industries
Fishes
Food Handling - methods
Food industries
Food Preservation - methods
Fundamental and applied biological sciences. Psychology
Glyceraldehyde-3-Phosphate Dehydrogenases - genetics
Marine
Molecular Sequence Data
Myosins - metabolism
Papain - metabolism
Pichia
Pichia pastoris
Promoter Regions, Genetic
Recombinant Proteins - biosynthesis
Recombinant Proteins - metabolism
Reverse Transcriptase Polymerase Chain Reaction
Scombridae
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Summary:A high level of the secreted form of recombinant chicken cystatin was expressed in Pichia pastoris X-33 by chromosomal integration of multiple copies of an expression cassette containing chicken cystatin under the control of glyceraldehyde-3-phosphate dehydrogenase promoter. The inhibition ability of the recombinant for papain-like proteinase was found to correspond to those of natural chicken cystatin. The recombinant cystatin substantially inhibited the proteolysis of myosin and gel softening, which consequently improved the gel properties of mackerel surimi. Keywords: Recombinant cystatin; mackerel surimi; Pichia pastoris; expression; cysteine proteinase inhibitor
ISSN:0021-8561
1520-5118
DOI:10.1021/jf000992s