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Klebsiella pneumoniae yggG gene product: a zinc-dependent metalloprotease


Kuan, Chee Sian and Wong, Mun Teng and Choi, Sy Bing and Chang, Ching Ching and Yoke, Hiang Yee and Abdul Wahab, Habibah and Mohd Yahaya, Normi and Wei, Cun See Too and Few, Ling Ling (2011) Klebsiella pneumoniae yggG gene product: a zinc-dependent metalloprotease. International Journal of Molecular Sciences, 12 (7). pp. 4441-4455. ISSN 1661-6596; ESSN: 1422-0067


Klebsiella pneumoniae causes neonatal sepsis and nosocomial infections. One of the strains, K. pneumoniae MGH 78578, shows high level of resistance to multiple microbial agents. In this study, domain family, amino acid sequence and topology analyses were performed on one of its hypothetical protein, YggG (KPN_03358). Structural bioinformatics approaches were used to predict the structure and functionality of YggG protein. The open reading frame (ORF) of yggG, which was a putative metalloprotease gene, was also cloned, expressed and characterized. The ORF was PCR amplified from K. pneumoniae MGH 78578 genomic DNA and cloned into a pET14-b vector for heterologous expression in Escherichia coli. The purified YggG protein was subsequently assayed for casein hydrolysis under different conditions. This protein was classified as peptidase M48 family and subclan gluzincin. It was predicted to contain one transmembrane domain by TMpred. Optimal protein expression was achieved by induction with 0.6 mM isopropyl thiogalactoside (IPTG) at 25 °C for six hours. YggG was purified as soluble protein and confirmed to be proteolytically active under the presence of 1.25 mM zinc acetate and showed optimum activity at 37 °C and pH 7.4. We confirmed for the first time that the yggG gene product is a zinc-dependent metalloprotease.

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Additional Metadata

Item Type: Article
Divisions: Faculty of Biotechnology and Biomolecular Sciences
DOI Number: https://doi.org/10.3390/ijms12074441
Publisher: Multidisciplinary Digital Publishing Institute
Keywords: Klebsiella pneumoniae MGH 78578; yggG gene; KPN_03358; Metalloprotease; HEXXH motif
Depositing User: Nurul Ainie Mokhtar
Date Deposited: 01 Sep 2015 06:12
Last Modified: 01 Sep 2015 06:12
Altmetrics: http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.3390/ijms12074441
URI: http://psasir.upm.edu.my/id/eprint/22358
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