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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Universiti Putra Malaysia
2019
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| Online Access: | http://irep.iium.edu.my/72576/ http://irep.iium.edu.my/72576/ 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 |
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iium-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 |
| repository_type |
Digital Repository |
| institution_category |
Local University |
| institution |
International Islamic University Malaysia |
| building |
IIUM Repository |
| collection |
Online Access |
| 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/ http://irep.iium.edu.my/72576/ 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 |
| first_indexed |
2023-09-18T21:42:50Z |
| last_indexed |
2023-09-18T21:42:50Z |
| _version_ |
1777413254525485056 |