Expression and identification of a minor extracellular fibrinolytic enzyme (Vpr) from Bacillus subtilis KCTC 3014 = Bacillus subtilis KCTC3014 유래 세포 외 분비형 혈전용해 부효소의 동정 및 발현

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Title
Expression and identification of a minor extracellular fibrinolytic enzyme (Vpr) from Bacillus subtilis KCTC 3014 = Bacillus subtilis KCTC3014 유래 세포 외 분비형 혈전용해 부효소의 동정 및 발현
Author(s)
Nack-Shick Choi; Dong-Min Jeong; Chan Sun Park; Keug Hyun AhnJoong Su KimJae Jun Song; Seung Ho Kim; Byung Dae Yoon; Min-Soo Kim
Bibliographic Citation
Biotechnology and Bioprocess Engineering, vol. 15, no. 3, pp. 446-452
Publication Year
2010
Abstract
Previously, three extracellular proteases, Vpr, PepT, and subtilisin were identified from Bacillus subtilis KCTC 3014. To confirm the activity of Vpr, two recombinant Vpr proteins, full Vpr with TTG (pGST-fTTG-Vpr) and full Vpr with ATG (pGST-fATG-Vpr) as an initiation codon were expressed using a pGEX-2T vector encoding glutathione S-transferase (GST) in Escherichia coli. Vpr was produced in two forms, occurring as four spots on a 2- DE gel, 68 and 75 kDa proteins with similar pI values (4.0 ? 4.5). Activity was detected in a fibrin zymography at the expected molecular size of 68 kDa (mature form) processed from full Vpr. However, the recombinant 75 kDa of GST-fVpr did not exhibit activity. Replacement of the TTG codon with ATG led to 1.9-fold increased enzyme activity in 68 kDa. Interestingly, the expression of GSTVpr resulted in the proteolytic degradation of the protein and no GST fusion Vpr protein was detected.
Keyword
Bacillus subtilisMass spectrometryUUG start codonVprZymography
ISSN
1226-8372
Publisher
Springer
DOI
http://dx.doi.org/10.1007/s12257-009-0191-z
Type
Article
Appears in Collections:
Jeonbuk Branch Institute > Immunoregulatory materials Research Center > 1. Journal Articles
Jeonbuk Branch Institute > Functional Biomaterial Research Center > 1. Journal Articles
Division of Bio Technology Innovation > SME Support Center > 1. Journal Articles
Jeonbuk Branch Institute > Microbial Biotechnology Research Center > 1. Journal Articles
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