Journal of Dairy Science and Biotechnology
- Volume 29 Issue 1
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- Pages.49-54
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- 2011
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- 2733-4554(pISSN)
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- 2733-4562(eISSN)
Cloning and Characterization of a Bile Salt Hydrolase from Enterococcus faecalis Strain Isolated from Healthy Elderly Volunteers
사람 분변에서 분리한 Enterococcusfaecalis가 생성하는 BileSaltHydrolase의 특징
- Eom, Seok-Jin (Dept. of Animal Science and Technology, Chung-Ang University) ;
- Kim, Geun-Bae (Dept. of Animal Science and Technology, Chung-Ang University)
- Received : 2011.06.10
- Accepted : 2011.06.27
- Published : 2011.06.30
Abstract
Bile salt hydrolase (BSH, EC 3.5.1.24) activity, which cleaves amide bond between carboxyl group (bile acid) and amino group (glycine or taurine), is commonly detected in gut-associated species of human and animal. During the screening of BSH active strains from the fecal samples of elderly human volunteers, strain CU30-2 was isolated on the basis of the highly active BSH producing activity. A bsh gene of the isolate was cloned into the pET22b expression vector and overexpressed in Escherichia coli BL21 (DE3) Gold by induction with 1mM IPTG. The overexpressed BSH enzyme with 6x His-tag was purified with apparent homogeneity using a
담즙산 분해효소(Bile salt hydrolase, EC 3.5.1.24) 활성은 담즙산의 카르복실기와 결합되어 있는 아미노기(glycine or taurine)와의 amide 결합을 끊는 작용을 하며, 이 효소 활성은 사람이나 동물의 장내 미생물들에서 널리 분포하고 있다. 노인 분변으로부터 분리한 여러 균주의 Enterococcus faecalis중에서 BSH activity가 가장 높은 CU30-2를 선발하였다. BSH 유전자를 pET22b expression vector에 클로닝하여 Escherichia coli BL21(DE3) Gold를 이용하여 단백질을 발현하였다. 6x His-tag이 있는 BSH 효소를