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Miniscale Identification and Characterization of Subtilisins from Bacillus sp. Strains

Journal of microbiology and biotechnology v.15 no.3 , 2005년, pp.537 - 543  
CHOI NACK-SHICK (Proteome Research Laboratory, Korea Research Institute of Bioscience and Biotechnology ) ;  JU SUNG-KYU ( Department of Biology, Chungnam National University ) ;  LEE TAE YOUNG ( Proteome Research Laboratory, Korea Research Institute of Bioscience and Biotechnology ) ;  YOON KAB-SEOG ( Proteome Research Laboratory, Korea Research Institute of Bioscience and Biotechnology ) ;  CHANG KYU-TAE ( Primate Research Laboratory, Korea Research Institute of Bioscience and Biotechnology ) ;  MAENG PIL JAE ( Microbiology, Chungnam National University ) ;  KIM SEUNG-HO ( Proteome Research Laboratory, Korea Research Institute of Bioscience and Biotechnology)
Abstract

Subtilisin (EC 3.4.21.14) is the major extracellular alkaline serine protease of Bacillus species. Previously, we found that subtilisins did not migrate in the electrophoretic field in the Laemmili buffer system due to their high pI values (over 8.8); however, it formed a 'binding mode' at the top of the separating gel [5]. Utilizing this characteristic, four subtilisins from Bacillus sp. strains (e.g., B. subtilis 168, B. subtilis KCTC 1021, B. amyloliquefaciens KCTC 3002, and Bacillus sp. DJ-1 and DJ-4) were easily and quickly identified by an over-running electrophoretic technique with a miniscale culture supernatant (less than 20 ml) without any column chromatographic steps. Two subtilisins (DJ-l and a recombinant version) from Bacillus sp. DJ-l were characterized, and the enzymatic properties were determined by SDS-fibrin zymography and densitometric analysis. Based on this observation, the recombinant pro-subtilisin DJ-l showed the same 'binding mode,' similar to native subtilisin DJ-l. On the other hand, mature subtilisin DJ -1 without pro-peptide showed no enzymatic activity.

주제어

#Subtilisin   #binding mode   #Bacillus   #over-running electrophoresis   #zymography  

저자의 다른 논문

최낙식(7) Yoon, Kab-Seog(7) Chang, Kyu-Tae(11) Maeng, Pil Jae(26) Kim, Seung-Ho(16)

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