Purification and biochemical characterization of Trypsin-like enzyme from Antarctic Hydrobiont Adamussium Colbecki

被引：0

作者：

Raksha, Nataliia ^{[1
]}

Halenova, Tetiana ^{[1
]}

Kravchenko, Olga ^{[1
]}

Vovk, Tetiana ^{[1
]}

Savchuk, Olexiy ^{[1
]}

Ostapchenko, Lydmila ^{[1
]}

机构：

[1] Taras Shevchenko Natl Univ, Inst Biol & Med, Educ & Sci Ctr, 64-13 Volodymyrska Str, UA-01601 Kiev, Ukraine

来源：

RESEARCH JOURNAL OF BIOTECHNOLOGY | 2020年 / 15卷 / 01期

关键词：

Hydrobiont; trypsin-like enzyme; isolation; biochemical properties; MOLECULAR-BASIS; PYLORIC CECA; LEPIDOPTERA; PROTEASES; PROTEINS; SMITH;

D O I：

暂无

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

A two-step high-yielding procedure was described for the purification of a trypsin-like enzyme from hydrobiont Adamussium colbecki, consisting of affinity chromatography on SBTI-sepharose 4B followed by size exclusion chromatography on Superdex 75 PG column. After the final purification step, trypsin-like enzyme was purified 7.52-fold with 50.3 % yield. The purified enzyme (24 kDa) was homogeneous as confirmed by SDS-PAGE. The pH and temperature optimum were estimated to be 9.0 and 24 degrees C. Trypsin-like enzyme was unstable above pH 12.0 and below pH 5.0. The enzyme was partially inhibited by PMSF, fully inhibited by SBTI and was not inhibited by EDTA, characterizing it as serine protease. The trypsin-like enzyme kinetic constant Km was lower at 8 degrees C compared to result at 24 degrees C while the catalytic efficiency kcat/Km was the same at both temperatures.

引用

页码：62 / 68

页数：7

共 23 条

[1]

ASGEIRSSON B, 1989, EUR J BIOCHEM, V180, P85

[2]

Bradford MM, 1976, Anal Biochem, V86, P193

[3] Epibionts of the scallop Adamussium colbecki (Smith, 1902) in the Ross Sea, Antarctica [J].