Affinity Chromatographic Puriﬁcation and Enzymatic Characterization of Trypsin from Grass Carp (Ctenopharyngodon idellus)

doi:10.7506/spkx1002-6630-20200304-062

FOOD SCIENCE ›› 2021, Vol. 42 ›› Issue (10): 178-183.doi: 10.7506/spkx1002-6630-20200304-062

• Bioengineering • Previous Articles Next Articles

Affinity Chromatographic Puriﬁcation and Enzymatic Characterization of Trypsin from Grass Carp (Ctenopharyngodon idellus)

LI Chen, GAO Liufang, CUI Xiaodong, HAN Yuhang, ZHU Danxu, LI Jiao

(1. School of Life Science, Shanxi University, Taiyuan 030006, China; 2. Institute of Biochemistry, Shanxi University, Taiyuan 030006, China)

Online:2021-05-25 Published:2021-06-02

Abstract

Abstract: Trypsin was extracted from the hepatopancreas of grass carps in this study. After homogenization and ammonium sulfate precipitation, crude enzyme solution was prepared by dialysis. Then, trypsin of electrophoretic purity was obtained by BTI-Sepharose affinity chromatography in only one step. The results showed that the molecular mass of the trypsin was about 27 kDa, and the Km was 3.4 × 10-5 mol/L. The optimum reaction temperature of the enzyme was 60 ℃ and it remained stabile at temperatures lower than 60 ℃. Its optimal reaction pH was 9.5, and the pH tolerance range was 6.0–12.0. Ba2+, Mg2+ and Fe2+ all could activate the enzyme in the concentration range of 0–10 mmol/L. EDTA and K+ inhibited the enzyme in the concentration range of 0–10 mmol/L, the former being more effective than the latter.

Key words: grass carp; trypsin; affinity chromatographic purification; enzymatic properties

CLC Number:

TS264.2

LI Chen, GAO Liufang, CUI Xiaodong, HAN Yuhang, ZHU Danxu, LI Jiao. Affinity Chromatographic Puriﬁcation and Enzymatic Characterization of Trypsin from Grass Carp (Ctenopharyngodon idellus)[J]. FOOD SCIENCE, 2021, 42(10): 178-183.

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Affinity Chromatographic Puriﬁcation and Enzymatic Characterization of Trypsin from Grass Carp (Ctenopharyngodon idellus)

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