Klebsiella pneumoniae yggG gene product: a zinc-dependent metalloprotease
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,...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Multidisciplinary Digital Publishing Institute
2011
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| Online Access: | http://psasir.upm.edu.my/id/eprint/22358/ http://psasir.upm.edu.my/id/eprint/22358/1/Klebsiella%20pneumoniae%20yggG%20Gene%20Product.pdf |
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| author | Kuan, Chee Sian Wong, Mun Teng Choi, Sy Bing Chang, Ching Ching Yoke, Hiang Yee Abdul Wahab, Habibah Mohd Yahaya, Normi Wei, Cun See Too Few, Ling Ling |
| author_facet | Kuan, Chee Sian Wong, Mun Teng Choi, Sy Bing Chang, Ching Ching Yoke, Hiang Yee Abdul Wahab, Habibah Mohd Yahaya, Normi Wei, Cun See Too Few, Ling Ling |
| author_sort | Kuan, Chee Sian |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | 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. |
| first_indexed | 2025-11-15T08:31:20Z |
| format | Article |
| id | upm-22358 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T08:31:20Z |
| publishDate | 2011 |
| publisher | Multidisciplinary Digital Publishing Institute |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-223582015-09-01T06:12:52Z http://psasir.upm.edu.my/id/eprint/22358/ Klebsiella pneumoniae yggG gene product: a zinc-dependent metalloprotease Kuan, Chee Sian Wong, Mun Teng Choi, Sy Bing Chang, Ching Ching Yoke, Hiang Yee Abdul Wahab, Habibah Mohd Yahaya, Normi Wei, Cun See Too Few, Ling Ling 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. Multidisciplinary Digital Publishing Institute 2011 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/22358/1/Klebsiella%20pneumoniae%20yggG%20Gene%20Product.pdf 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 http://www.mdpi.com/1422-0067/12/7/4441?trendmd-shared=0 10.3390/ijms12074441 |
| spellingShingle | Kuan, Chee Sian Wong, Mun Teng Choi, Sy Bing Chang, Ching Ching Yoke, Hiang Yee Abdul Wahab, Habibah Mohd Yahaya, Normi Wei, Cun See Too Few, Ling Ling Klebsiella pneumoniae yggG gene product: a zinc-dependent metalloprotease |
| title | Klebsiella pneumoniae yggG gene product: a zinc-dependent metalloprotease |
| title_full | Klebsiella pneumoniae yggG gene product: a zinc-dependent metalloprotease |
| title_fullStr | Klebsiella pneumoniae yggG gene product: a zinc-dependent metalloprotease |
| title_full_unstemmed | Klebsiella pneumoniae yggG gene product: a zinc-dependent metalloprotease |
| title_short | Klebsiella pneumoniae yggG gene product: a zinc-dependent metalloprotease |
| title_sort | klebsiella pneumoniae yggg gene product: a zinc-dependent metalloprotease |
| url | http://psasir.upm.edu.my/id/eprint/22358/ http://psasir.upm.edu.my/id/eprint/22358/ http://psasir.upm.edu.my/id/eprint/22358/ http://psasir.upm.edu.my/id/eprint/22358/1/Klebsiella%20pneumoniae%20yggG%20Gene%20Product.pdf |