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Partial purification and model structure of BPSL2774, a hypothetical protein from Burkholderia pseudomallei predicted to be a Glycosyltransferase

Melioidosis is a disease that infects humans and animals, and can be detrimental in humans. Mortality rate from melioidosis septic shock due to infection from Gram negative Burkholderia pseudomallei (B. pseudomallei) in endemic regions of Malaysia and Thailand remains high despite available antim...

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Main Authors: Drahaman, Siti Marhamah, Ujang, Hanisah, Mat Akhir, Nor Azurah, Muhamad Bunnori, Noraslinda, Mohamed Rehan, Aisyah
Format: Article
Language:English
English
Published: Universiti Putra Malaysia 2019
Subjects:
QR Microbiology
TP248.13 Biotechnology
Online Access:http://irep.iium.edu.my/72576/1/15%20JTAS-1245-2017.pdf
http://irep.iium.edu.my/72576/7/72576_Partial%20Purification%20and%20Model%20Structure_scopus.pdf
http://irep.iium.edu.my/72576/
http://www.pertanika.upm.edu.my/Pertanika%20PAPERS/JTAS%20Vol.%2042%20(2)%20May.%202019/15%20JTAS-1245-2017.pdf
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spelling my.iium.irep.725762019-11-24T13:38:20Z http://irep.iium.edu.my/72576/ Partial purification and model structure of BPSL2774, a hypothetical protein from Burkholderia pseudomallei predicted to be a Glycosyltransferase Drahaman, Siti Marhamah Ujang, Hanisah Mat Akhir, Nor Azurah Muhamad Bunnori, Noraslinda Mohamed Rehan, Aisyah QR Microbiology TP248.13 Biotechnology Melioidosis is a disease that infects humans and animals, and can be detrimental in humans. Mortality rate from melioidosis septic shock due to infection from Gram negative Burkholderia pseudomallei (B. pseudomallei) in endemic regions of Malaysia and Thailand remains high despite available antimicrobial therapy. Multiple strategies are employed to identify essential genes and drug targets in this bacterium to improve current antimicrobial therapies. This is important as B. pseudomallei is intrinsically resistant to many commonly used antibiotics. In this study, hypothetical genes predicted to be essential for B. pseudomallei by transposon-directed insertion site sequencing (TraDIS) technique were selected. One target gene, BPSL2774, has been successfully amplified and cloned from genomic DNA of B. pseudomallei strain K96243. Glutathione S-transferase (GST) affinity tag chromatography was performed for partial protein purification. The target protein was successfully expressed in soluble form with satisfactory yield output. Mass spectrometry analysis of 60 kDa Coomassie-stained gel band confirmed the presence of the soluble expressed taggedtarget protein, co-purified with Escherichia coli chaperonin proteins, possibly due to their interaction with the target protein. BPSL2774 protein have considerable homology to glycosyltransferase GTB type superfamily and RfaB superfamily. On the basis of this similarity, the three-dimensional structure of BPSL2774 has been modelled and assessed by protein model quality servers. Taking all the results into account, the functional annotation of BPSL2774 protein as a glycosyltransferase is recommended, though future validation from biochemical experiments will be needed to support this. Universiti Putra Malaysia 2019-05 Article PeerReviewed application/pdf en http://irep.iium.edu.my/72576/1/15%20JTAS-1245-2017.pdf application/pdf en http://irep.iium.edu.my/72576/7/72576_Partial%20Purification%20and%20Model%20Structure_scopus.pdf Drahaman, Siti Marhamah and Ujang, Hanisah and Mat Akhir, Nor Azurah and Muhamad Bunnori, Noraslinda and Mohamed Rehan, Aisyah (2019) Partial purification and model structure of BPSL2774, a hypothetical protein from Burkholderia pseudomallei predicted to be a Glycosyltransferase. Journal of Tropical Agricultural Science, 42 (2). pp. 609-625. ISSN 1511-3701 E-ISSN 2231-8542 http://www.pertanika.upm.edu.my/Pertanika%20PAPERS/JTAS%20Vol.%2042%20(2)%20May.%202019/15%20JTAS-1245-2017.pdf
institution Universiti Islam Antarabangsa Malaysia
building IIUM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider International Islamic University Malaysia
content_source IIUM Repository (IREP)
url_provider http://irep.iium.edu.my/
language English
English
topic QR Microbiology
TP248.13 Biotechnology
spellingShingle QR Microbiology
TP248.13 Biotechnology
Drahaman, Siti Marhamah
Ujang, Hanisah
Mat Akhir, Nor Azurah
Muhamad Bunnori, Noraslinda
Mohamed Rehan, Aisyah
Partial purification and model structure of BPSL2774, a hypothetical protein from Burkholderia pseudomallei predicted to be a Glycosyltransferase
description Melioidosis is a disease that infects humans and animals, and can be detrimental in humans. Mortality rate from melioidosis septic shock due to infection from Gram negative Burkholderia pseudomallei (B. pseudomallei) in endemic regions of Malaysia and Thailand remains high despite available antimicrobial therapy. Multiple strategies are employed to identify essential genes and drug targets in this bacterium to improve current antimicrobial therapies. This is important as B. pseudomallei is intrinsically resistant to many commonly used antibiotics. In this study, hypothetical genes predicted to be essential for B. pseudomallei by transposon-directed insertion site sequencing (TraDIS) technique were selected. One target gene, BPSL2774, has been successfully amplified and cloned from genomic DNA of B. pseudomallei strain K96243. Glutathione S-transferase (GST) affinity tag chromatography was performed for partial protein purification. The target protein was successfully expressed in soluble form with satisfactory yield output. Mass spectrometry analysis of 60 kDa Coomassie-stained gel band confirmed the presence of the soluble expressed taggedtarget protein, co-purified with Escherichia coli chaperonin proteins, possibly due to their interaction with the target protein. BPSL2774 protein have considerable homology to glycosyltransferase GTB type superfamily and RfaB superfamily. On the basis of this similarity, the three-dimensional structure of BPSL2774 has been modelled and assessed by protein model quality servers. Taking all the results into account, the functional annotation of BPSL2774 protein as a glycosyltransferase is recommended, though future validation from biochemical experiments will be needed to support this.
format Article
author Drahaman, Siti Marhamah
Ujang, Hanisah
Mat Akhir, Nor Azurah
Muhamad Bunnori, Noraslinda
Mohamed Rehan, Aisyah
author_facet Drahaman, Siti Marhamah
Ujang, Hanisah
Mat Akhir, Nor Azurah
Muhamad Bunnori, Noraslinda
Mohamed Rehan, Aisyah
author_sort Drahaman, Siti Marhamah
title Partial purification and model structure of BPSL2774, a hypothetical protein from Burkholderia pseudomallei predicted to be a Glycosyltransferase
title_short Partial purification and model structure of BPSL2774, a hypothetical protein from Burkholderia pseudomallei predicted to be a Glycosyltransferase
title_full Partial purification and model structure of BPSL2774, a hypothetical protein from Burkholderia pseudomallei predicted to be a Glycosyltransferase
title_fullStr Partial purification and model structure of BPSL2774, a hypothetical protein from Burkholderia pseudomallei predicted to be a Glycosyltransferase
title_full_unstemmed Partial purification and model structure of BPSL2774, a hypothetical protein from Burkholderia pseudomallei predicted to be a Glycosyltransferase
title_sort partial purification and model structure of bpsl2774, a hypothetical protein from burkholderia pseudomallei predicted to be a glycosyltransferase
publisher Universiti Putra Malaysia
publishDate 2019
url http://irep.iium.edu.my/72576/1/15%20JTAS-1245-2017.pdf
http://irep.iium.edu.my/72576/7/72576_Partial%20Purification%20and%20Model%20Structure_scopus.pdf
http://irep.iium.edu.my/72576/
http://www.pertanika.upm.edu.my/Pertanika%20PAPERS/JTAS%20Vol.%2042%20(2)%20May.%202019/15%20JTAS-1245-2017.pdf
_version_ 1651865919842942976
score 13.211869

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