FRAGMENTATION AND CHARACTERISTICS OF RECOMBINANT ENDOCHITINASE EXPRESSED IN ESCHERICHIA COLI

被引：0
作者：
Chen, Wei-Ming [1 ,2 ]
Chen, Gen-Hung [3 ]
Jiang, Shann-Tzong [1 ,4 ]
机构：
[1] Natl Taiwan Ocean Univ, Dept Food Sci, Keelung, Taiwan
[2] Taiwan Agr Chem & Tox Subst Res Inst, Taichung, Taiwan
[3] Providence Univ, Dept Cosmet Sci, Taichung, Taiwan
[4] Providence Univ, Dept Food & Nutr, Taichung, Taiwan
来源：
JOURNAL OF MARINE SCIENCE AND TECHNOLOGY-TAIWAN | 2012年 / 20卷 / 04期
关键词：
expression; endochitinase; Escherichia coli; proteolysis; BIOCONTROL AGENT; BACILLUS-CEREUS; CELL ATTACHMENT; CHITINASE; ENZYMES; PURIFICATION; EXTRACTION; CONVERSION; CLONING; GENES;
D O I：
10.6119/JMST-011-0225-1
中图分类号：
T [工业技术];
学科分类号：
08 ;
摘要：
A recombinant endochitinase fused with hexahistidine (rEC-H) was expressed and characterized. After Ni affinity chromatography, the recovery, purification-fold and specific activity of rEC-H were 88.8%, 13.4 and 142.1 U/mg, respectively. The purified rEC-H had optimal pH and temperature at pH 7.5 and 60 degrees C, respectively and was stable at pH 4.0-9.0 and <50 degrees C. It was activated by Ca2+, Sr2+, Ba2+, Co2+ and beta-mercaptoethanol, but highly inhibited by Cu2+, Hg2+ and SDS. The thiol group may play an important role on rEC-H and rEC-H may easily be destructed when hydrophobic interaction is disrupted by sodium dodecyl sulfate. The molecular mass was lower than that of expected clue to cleavage at 2 specific sites, (33)Ala-(34) Asp and (472)Glu-(473)Leu within rEC-H, suggesting the fragmentation of rEC-H occurred during expression.
引用
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页码：369 / 375
页数：7
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