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/ http://irep.iium.edu.my/47160/ http://irep.iium.edu.my/47160/ http://irep.iium.edu.my/47160/1/Paper_-_Nuge_et_al_2014_%28Adv_in_Enzyme_Res%29.pdf

id	iium-47160
recordtype	eprints
spelling	iium-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 PeerReviewed 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 E-ISSN 2328-4854 http://www.scirp.org/Journal/PaperInformation.aspx?PaperID=43904 10.4236/aer.2014.21003
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
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/ http://irep.iium.edu.my/47160/ http://irep.iium.edu.my/47160/ http://irep.iium.edu.my/47160/1/Paper_-_Nuge_et_al_2014_%28Adv_in_Enzyme_Res%29.pdf
first_indexed	2023-09-18T21:07:08Z
last_indexed	2023-09-18T21:07:08Z
_version_	1777411008220889088

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

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