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Cloning and expression of a novel phytase gene (phyMS) from Mycobacterium smegmatis

Phytase, also known as phytate-degrading enzyme, catalyzes the hydrolysis of phytate (inositol hexakisphosphate) with sequential release of phosphate and lower inositol phosphate. We report here a new plasmid construct designated as pMSuia from pBAD-TOPO that harbors a 1.1 kb phytase gene (phyMS)...

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Main Authors: Nuge, Tamrin, Hashim, Yumi Zuhanis Has-Yun, Farouk, Abd-El Alziem, Mohd. Salleh, Hamzah
Format: Article
Language:English
Published: Scientific Research Publishing Inc. 2014
Subjects:
Q Science (General)
Online Access:http://irep.iium.edu.my/47160/1/Paper_-_Nuge_et_al_2014_%28Adv_in_Enzyme_Res%29.pdf
http://irep.iium.edu.my/47160/
http://www.scirp.org/Journal/PaperInformation.aspx?PaperID=43904
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spelling my.iium.irep.471602018-01-04T03:37:14Z http://irep.iium.edu.my/47160/ Cloning and expression of a novel phytase gene (phyMS) from Mycobacterium smegmatis Nuge, Tamrin Hashim, Yumi Zuhanis Has-Yun Farouk, Abd-El Alziem Mohd. Salleh, Hamzah Q Science (General) Phytase, also known as phytate-degrading enzyme, catalyzes the hydrolysis of phytate (inositol hexakisphosphate) with sequential release of phosphate and lower inositol phosphate. We report here a new plasmid construct designated as pMSuia from pBAD-TOPO that harbors a 1.1 kb phytase gene (phyMS) from Mycobacterium smegmatis, and expression as well as characterization of the purified recombinant M. smegmatis phytase. DNA sequencing analysis and multiple alignment exercise indicated that the M. smegmatis phytase is different from both known acid and alkaline phytases. The active ~45 kDa recombinant enzyme was expressed and confirmed by enzyme assay and Western blot analyses. Ni-NTA affinity purified recombinant M. smegmatis phytase exhibited specific activity of 233.51 U/mg, optimal pH of 3 and 7 and optimal temperature of 60˚C. The purified enzyme retains almost 30% of the initial activity after incubation at 90˚C for 60 min. The enzyme showed broad substrate specificity with Km and Vmax of the recombinant enzyme for sodium phytate substrate of 0.105 ± 0.016 mM and 26.93 ± 1.21 mM min−1, respectively. Scientific Research Publishing Inc. 2014-03 Article REM application/pdf en http://irep.iium.edu.my/47160/1/Paper_-_Nuge_et_al_2014_%28Adv_in_Enzyme_Res%29.pdf Nuge, Tamrin and Hashim, Yumi Zuhanis Has-Yun and Farouk, Abd-El Alziem and Mohd. Salleh, Hamzah (2014) Cloning and expression of a novel phytase gene (phyMS) from Mycobacterium smegmatis. Advances in Enzyme Research, 2 (1). pp. 27-38. ISSN 2328-4846 http://www.scirp.org/Journal/PaperInformation.aspx?PaperID=43904 10.4236/aer.2014.21003
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
topic Q Science (General)
spellingShingle Q Science (General)
Nuge, Tamrin
Hashim, Yumi Zuhanis Has-Yun
Farouk, Abd-El Alziem
Mohd. Salleh, Hamzah
Cloning and expression of a novel phytase gene (phyMS) from Mycobacterium smegmatis
description Phytase, also known as phytate-degrading enzyme, catalyzes the hydrolysis of phytate (inositol hexakisphosphate) with sequential release of phosphate and lower inositol phosphate. We report here a new plasmid construct designated as pMSuia from pBAD-TOPO that harbors a 1.1 kb phytase gene (phyMS) from Mycobacterium smegmatis, and expression as well as characterization of the purified recombinant M. smegmatis phytase. DNA sequencing analysis and multiple alignment exercise indicated that the M. smegmatis phytase is different from both known acid and alkaline phytases. The active ~45 kDa recombinant enzyme was expressed and confirmed by enzyme assay and Western blot analyses. Ni-NTA affinity purified recombinant M. smegmatis phytase exhibited specific activity of 233.51 U/mg, optimal pH of 3 and 7 and optimal temperature of 60˚C. The purified enzyme retains almost 30% of the initial activity after incubation at 90˚C for 60 min. The enzyme showed broad substrate specificity with Km and Vmax of the recombinant enzyme for sodium phytate substrate of 0.105 ± 0.016 mM and 26.93 ± 1.21 mM min−1, respectively.
format Article
author Nuge, Tamrin
Hashim, Yumi Zuhanis Has-Yun
Farouk, Abd-El Alziem
Mohd. Salleh, Hamzah
author_facet Nuge, Tamrin
Hashim, Yumi Zuhanis Has-Yun
Farouk, Abd-El Alziem
Mohd. Salleh, Hamzah
author_sort Nuge, Tamrin
title Cloning and expression of a novel phytase gene (phyMS) from Mycobacterium smegmatis
title_short Cloning and expression of a novel phytase gene (phyMS) from Mycobacterium smegmatis
title_full Cloning and expression of a novel phytase gene (phyMS) from Mycobacterium smegmatis
title_fullStr Cloning and expression of a novel phytase gene (phyMS) from Mycobacterium smegmatis
title_full_unstemmed Cloning and expression of a novel phytase gene (phyMS) from Mycobacterium smegmatis
title_sort cloning and expression of a novel phytase gene (phyms) from mycobacterium smegmatis
publisher Scientific Research Publishing Inc.
publishDate 2014
url http://irep.iium.edu.my/47160/1/Paper_-_Nuge_et_al_2014_%28Adv_in_Enzyme_Res%29.pdf
http://irep.iium.edu.my/47160/
http://www.scirp.org/Journal/PaperInformation.aspx?PaperID=43904
_version_ 1643613138645417984
score 13.211869

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