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dc.contributor.authorWang, Qin
dc.contributor.author王勤
dc.contributor.authorGao, Zhi-Xing
dc.contributor.authorShi, Yan
dc.contributor.author石艳
dc.contributor.authorChen, Qing-Xi
dc.contributor.author陈清西
dc.contributor.authorZhang, Nong
dc.contributor.authorXie, Xiao-Lan(Quangzhou Normal Univ)
dc.date.accessioned2011-07-13T05:56:07Z
dc.date.available2011-07-13T05:56:07Z
dc.date.issued2010
dc.identifier.citationJ. Agric. Food Chem., 2010, 58 (1), pp 655–659zh_CN
dc.identifier.issn0021-8561
dc.identifier.urihttp://dx.doi.org/doi:10.1021/jf903052s
dc.identifier.urihttps://dspace.xmu.edu.cn/handle/2288/10136
dc.description.abstractTrypsin from the intestine of hybrid tilapia (Oreochromis niloticus x O.aureus) was purified by the following techniques: acetone precipitation, ammonium sulfate fractionation, Sephacryl S-200 gel filtration, and DEAE-sephacel ion exchange chromatography. The purified enzyme was determined to be homogeneous by polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl sulfate (SDS)-PAGE. The molecular weight was estimated as 22,000 Da. The optimum pH and temperature of the enzyme for the hydrolysis of casein were determined to be 9.0 and 60 degrees C, respectively. The enzyme was stable over a broad pH range from 7.0 to 12.0 at 30 degrees C, and the enzyme was inactive at temperatures above 50 degrees C. The behavior of the enzyme for the hydrolysis of casein followed Michaelis-Menten kinetics with K-m of 0.46 mg/mL. The purified enzyme was inhibited by the general serine protease inhibitor phenyl methyl sulphonyl fluoride (PMSF) and also by the specific trypsin inhibitor N-p-tosyl-L-lysine chloromethyl ketone (TLCK) using N-alpha-CBZ-L-lysine p-nitrophenyl ester hydrochloride (CBZ-Lys.pNP) as a substrate. The protease was inhibited by the following ions in decreasing order: Zn2+ > Fe3+ > Cu2+ > Al3+ > Co2+ = Pb2+ > Cd2+ > Mn2+. The ions Li+, Na+, K+, Mg2+, and Ba2+ had little effect on the enzyme, and Ca2+, can partially promote its activity at low concentration.zh_CN
dc.description.sponsorshipedu.cnzh_CN
dc.language.isoenzh_CN
dc.publisherAMER CHEMICAL SOCzh_CN
dc.subjectTrypsin; purificationzh_CN
dc.subjectcharacterizationzh_CN
dc.subjectintestinezh_CN
dc.subjecthybrid tilapiazh_CN
dc.titlePurification and Characterization of Trypsin from the Intestine of Hybrid Tilapia (Oreochromis niloticus x O.aureus)zh_CN
dc.typeArticlezh_CN


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