N-Terminal seven-amino-acid extension simultaneously improves the pH stability, optimal temperature, thermostability and catalytic efficiency of chitosanase CsnA

被引：9
|
作者：
Han, Yujuan [1 ,2 ,3 ,4 ]
Gao, Peixin [1 ,2 ,3 ,4 ]
Yu, Wengong [1 ,2 ,3 ,4 ]
Lu, Xinzhi [1 ,2 ,3 ,4 ]
机构：
[1] Ocean Univ China, Key Lab Glycosci & Glycotechnol Shandong Prov, Sch Med & Pharm, 5 Yushan Rd, Qingdao 266003, Peoples R China
[2] Ocean Univ China, Chinese Minist Educ, Key Lab Marine Drugs, Sch Med & Pharm, 5 Yushan Rd, Qingdao 266003, Peoples R China
[3] Ocean Univ China, Qingdao Natl Lab Marine Sci & Technol, Lab Marine Drugs & Bioprod, Sch Med & Pharm, 5 Yushan Rd, Qingdao 266003, Peoples R China
[4] Ocean Univ China, Sch Med & Pharm, Dept Glycobiol, 5 Yushan Rd, Qingdao 266003, Peoples R China
来源：
BIOTECHNOLOGY LETTERS | 2018年 / 40卷 / 01期
基金：
中国国家自然科学基金;
关键词：
Catalytic efficiency; Chitosanase; Kinetic parameters; N-terminal sequence; Thermostability; AMINO-ACIDS; GH10; XYLANASE; STABILIZATION; N174;
D O I：
10.1007/s10529-017-2436-9
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：
To determine the effects of the extra N-terminal seven-amino-acid sequence on the function of chitosanase CsnA. Sequence and structure analysis indicated that the mature CsnA contains a seven-amino-acid extension in a disordered form at the N-terminus. To determine the function of this sequence, both mature CsnA and its N-terminus-truncated mutant, CsnA Delta N, were expressed in Escherichia coli and characterized. Compared with CsnA Delta N, CsnA exhibited a 15 A degrees C higher temperature optimum, enhanced pH stability, thermostability and catalytic efficiency. The underlying mechanisms responsible for these changes were analyzed by circular dichroism (CD) spectroscopy. CD analysis revealed that the deletion of the N-terminal sequence resulted in a decrease in the T-m of 4.3 A degrees C and this sequence altered the secondary structure of the enzyme. The N-terminal sequence is essential for the stability and activity of chitosanase CsnA.
引用
收藏
页码：75 / 82
页数：8
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