Purification and characterization of cell-envelopeproteinase from Lactobacillus casei DI-1
Purification and characterization of cell-envelopeproteinase from Lactobacillus casei DI-1
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干酪乳杆菌 DI-1 细胞膜蛋白酶的纯化和表征
DOI：
10.5897/ajb11.1784
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发表时间：
2012-10
影响因子：
--
通讯作者：
潘道东
中科院分区：
文献类型：
--
作者：
郭宇星;潘道东
采用无钙法从鸭小肠分离出干酪乳杆菌DI-1的细胞膜蛋白酶（CEP），经硫酸铵沉淀、DEAE-SephadexA-25和SephadexG-100凝胶层析纯化。纯化的CEP具有单体结构，分子量约为35 kDa。最佳活性出现在pH 7.0和37°C。纯化的CEP是一种金属肽酶，能被Co2+、Ba 2+、Mg 2+和Fe 3+激活，被Ca 2+、Zn 2+、K +、Ni 2+、Mn 2+和乙二胺四乙酸（EDTA）抑制。它是一种丝氨酸蛋白酶，可被苯甲基磺酰氟（PMSF）抑制。酶解动力学常数Km为0.29 mM，N端前10个氨基酸序列为Asp-Asn-Asp-Phe-Glu-Ile-Phe-Glu-Ser-Ser，酶解产物对α-、β-和κ-酪蛋白的ACE抑制活性不同，β-酪蛋白酶解产物的ACE抑制活性最强。关键词：细胞膜蛋白酶，纯化，性质。
Using a Ca 2+ -free method, the cell-envelope proteinase (CEP) of Lactobacillus casei DI-1 isolated from duck small intestine was released from cells and purified by ammonium sulfate precipitation, and by diethylaminoethyl (DEAE)-Sephadex A-25 and Sephadex G-100 gel chromatography. The purified CEP had a monomer structure with a molecular mass of about 35 kDa. Optimal activity occurred at pH 7.0 and 37°C. The purified CEP was a metallopeptidase, which was activated by Co 2+ , Ba 2+ , Mg 2+ and Fe 3+ , and inhibited by Ca 2+ , Zn 2+ , K + , Ni 2+ , Mn 2+ , and ethylenediaminetetraacetic acid (EDTA). It was a serine proteinase which was inhibited by phenylmethylsulfonyl fluoride (PMSF). Its kinetic constant (Km) is 0.29 mM and the first 10 amino acids of the CEP’s N-terminal sequences were Asp-Asn-Asp-Phe-Glu-Ile-Phe-Glu-Ser-Ser. The hydrolysates of α-, β- and κ-casein produced by CEP showed different angiotensin-I-converting enzyme (ACE) inhibitory activity; the hydrolysates of β-casein displayed the greatest ACE inhibitory activity. Key words : Cell-envelope proteinase, purification, characterization.
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影响因子：
6
作者：
Sinsuwan, Sornchai;Rodtong, Sureelak;Yongsawatdigul, Hrawat
通讯作者：
Yongsawatdigul, Hrawat
影响因子：
3.1
作者：
W. Bockelmann
通讯作者：
W. Bockelmann
影响因子：
--
作者：
R. Mirnejad;J. Hossein;Abdolla Ardebilli;H. Babavalian
通讯作者：
R. Mirnejad;J. Hossein;Abdolla Ardebilli;H. Babavalian
影响因子：
4.4
作者：
KOJIC, M;FIRA, D;TOPISIROVIC, L
通讯作者：
TOPISIROVIC, L
影响因子：
3.5
作者：
N. Yamamoto;A. Akino;T. Takano
通讯作者：
N. Yamamoto;A. Akino;T. Takano