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Biochemical and molecular characterization of a novel calcium-dependent metalloprotease from Bacillus cereus SV1

Paper ID Volume ID Publish Year Pages File Format Full-Text
35634 45099 2008 9 PDF Available
Title
Biochemical and molecular characterization of a novel calcium-dependent metalloprotease from Bacillus cereus SV1
Abstract

An extracellular calcium-dependent protease from a newly isolated Bacillus cereus SV1 was purified, characterized and the gene was isolated and sequenced. The enzyme was purified to homogeneity by ultrafiltration, Sephacryl S-200 gel filtration, DEAE-cellulose ion exchange chromatography, then by a second gel filtration on Sephacryl S-200 with a six-fold increase in specific activity and 28% recovery. The production of SV1 protease was observed in media containing CaCl2 but not in media containing BaCl2, MgCl2, MnCl2 and ZnCl2. The molecular weight of the enzyme was estimated to be 35.5 kDa on SDS-PAGE. The enzyme exhibited maximal activity at pH 8.0 and 55 °C, without the presence of CaCl2. Thermoactivity and thermostability of the enzyme were significantly increased in the presence of Ca2+. The activity was totally lost in the presence of EDTA, suggesting that the purified enzyme is a metalloprotease.The nprC1 gene, which encodes the calcium-dependent protease from B. cereus SV1, was isolated and its DNA sequence was determined. The nprC1 gene consists of 1701 bp encoding a preproprotein of 566 amino acids with three distinct domains: a signal peptide (27 amino acids), a pro-peptide (222 amino acids) and a mature enzyme (317 amino acids). The nprC1 gene coding sequence presents only 91% and 89% identity to those encoding neutral protease from B. cereus ATCC 14579 and bacillolysin from B. cereus DSM 3101, respectively. The deduced amino acid sequence of the mature protease (NprC1) differs from those of DSM 3101 and ATCC 14579 with 16 and 17 amino acids, respectively, suggesting that the nprC1 is a new protease gene.

Keywords
Bacillus cereus; Metalloprotease; Calcium-dependent; Thermolysin-like; Purification; DNA sequence
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Biochemical and molecular characterization of a novel calcium-dependent metalloprotease from Bacillus cereus SV1
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 43, Issue 5, May 2008, Pages 522–530
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us