Expression, purification, and characterization of a recombinant stem bromelain from ananas comosus

Commercially available bromelain is prepared by performing a tedious and costly purification method that provides multiple degrees of purified bromelain. In the current study, a gene encoding stem bromelain from Ananas comosus was amplified using polymerase chain reaction (PCR). This bromelain gene...

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Main Authors:	Amid, Azura, Ismail, Nurul Azira, Yusof, Faridah, Mohd. Salleh, Hamzah
Format:	Article
Language:	English
Published:	ELSEVIER 2011
Subjects:	Q Science (General)
Online Access:	http://irep.iium.edu.my/2374/ http://irep.iium.edu.my/2374/ http://irep.iium.edu.my/2374/1/Bromelain_cloning.pdf

id	iium-2374
recordtype	eprints
spelling	iium-23742012-04-19T03:45:33Z http://irep.iium.edu.my/2374/ Expression, purification, and characterization of a recombinant stem bromelain from ananas comosus Amid, Azura Ismail, Nurul Azira Yusof, Faridah Mohd. Salleh, Hamzah Q Science (General) Commercially available bromelain is prepared by performing a tedious and costly purification method that provides multiple degrees of purified bromelain. In the current study, a gene encoding stem bromelain from Ananas comosus was amplified using polymerase chain reaction (PCR). This bromelain gene was initially cloned into pENTR/TEV/D-TOPO before being sub-cloned into the expression vector pDEST17. DNA sequencing of the amplified products exhibited a high level of homology to the corresponding gene from the NCBI public database. Protein expression was conducted in the Escherichia coli, BL21-AI. The recombinant bromelain was then purified in a single step using a Ni-NTA spin column, an example of immobilized metal affinity chromatography. Purified recombinant bromelain was detected by Western blotting. In addition, the purified enzyme exhibited hydrolytic activity towards gelatin and a synthetic substrate, LNPE. The purified recombinant bromelain exhibited optimum activity at pH 4.6 and 45oC. ELSEVIER 2011-09 Article PeerReviewed application/pdf en http://irep.iium.edu.my/2374/1/Bromelain_cloning.pdf Amid, Azura and Ismail, Nurul Azira and Yusof, Faridah and Mohd. Salleh, Hamzah (2011) Expression, purification, and characterization of a recombinant stem bromelain from ananas comosus. Process Biochemistry, 46 (12). pp. 2232-2239. ISSN 1359-5113 http://www.elsevier.com/wps/find/journaldescription.cws_home/422857/description#description
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) Amid, Azura Ismail, Nurul Azira Yusof, Faridah Mohd. Salleh, Hamzah Expression, purification, and characterization of a recombinant stem bromelain from ananas comosus
description	Commercially available bromelain is prepared by performing a tedious and costly purification method that provides multiple degrees of purified bromelain. In the current study, a gene encoding stem bromelain from Ananas comosus was amplified using polymerase chain reaction (PCR). This bromelain gene was initially cloned into pENTR/TEV/D-TOPO before being sub-cloned into the expression vector pDEST17. DNA sequencing of the amplified products exhibited a high level of homology to the corresponding gene from the NCBI public database. Protein expression was conducted in the Escherichia coli, BL21-AI. The recombinant bromelain was then purified in a single step using a Ni-NTA spin column, an example of immobilized metal affinity chromatography. Purified recombinant bromelain was detected by Western blotting. In addition, the purified enzyme exhibited hydrolytic activity towards gelatin and a synthetic substrate, LNPE. The purified recombinant bromelain exhibited optimum activity at pH 4.6 and 45oC.
format	Article
author	Amid, Azura Ismail, Nurul Azira Yusof, Faridah Mohd. Salleh, Hamzah
author_facet	Amid, Azura Ismail, Nurul Azira Yusof, Faridah Mohd. Salleh, Hamzah
author_sort	Amid, Azura
title	Expression, purification, and characterization of a recombinant stem bromelain from ananas comosus
title_short	Expression, purification, and characterization of a recombinant stem bromelain from ananas comosus
title_full	Expression, purification, and characterization of a recombinant stem bromelain from ananas comosus
title_fullStr	Expression, purification, and characterization of a recombinant stem bromelain from ananas comosus
title_full_unstemmed	Expression, purification, and characterization of a recombinant stem bromelain from ananas comosus
title_sort	expression, purification, and characterization of a recombinant stem bromelain from ananas comosus
publisher	ELSEVIER
publishDate	2011
url	http://irep.iium.edu.my/2374/ http://irep.iium.edu.my/2374/ http://irep.iium.edu.my/2374/1/Bromelain_cloning.pdf
first_indexed	2023-09-18T20:09:58Z
last_indexed	2023-09-18T20:09:58Z
_version_	1777407411648200704

Expression, purification, and characterization of a recombinant stem bromelain from ananas comosus

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