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 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.
Deaprtment of Food Science and Institute of Marine Biotechnology, National Taiwan Ocean University, Keelung, Taiwan.
G H
GH
Chen
S J
SJ
Tang
C S
CS
Chen
S T
ST
Jiang
D000818:Animals; D001483:Base Sequence; D002645:Chickens; D003001:Cloning, Molecular / Q000379:methods; D015891:Cystatins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D017931:DNA Primers; D010450:Endopeptidases / Q000378:metabolism; D005399:Fishes; D005511:Food Handling / Q000379:methods; D005519:Food Preservation / Q000379:methods*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D008969:Molecular Sequence Data; D009218:Myosins / Q000378:metabolism; D010206:Papain / Q000378:metabolism; D010843:Pichia; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D015891:Cystatins; D017931:DNA Primers; D011994:Recombinant Proteins; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D010450:Endopeptidases; D010206:Papain; D009218:Myosins