Cloning of pullulanase gene from local isolated bacteria

Pullulanases (pullulan-6-glucanohydrolase, EC 3.2.1.41) are debranching enzymes that are able to hydrolyze α-1,6-glycosidic linkages in pullulan and branched polysaccharides, producing maltotriose. Pullulanases are a member of family 13 glycosyl hydrolases or α-amylase family. Pullulanase are clas...

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Main Authors: Chee, Chit Lai, Md. Illias, Rosli, Hassan, Osman, Kamaruddin, Kamarulzaman, Abdul Aziz, Suraini, Md. Salleh, Madihah, Wan Md. Zain, Wan Salwanis
Format: Conference or Workshop Item
Language:English
Published: 2005
Subjects:
Online Access:http://eprints.utm.my/5370/
http://eprints.utm.my/5370/1/CheeChitLai2005_CloningOfPullulanaseGeneFromLocalIsolatedBacteria.pdf
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author Chee, Chit Lai
Md. Illias, Rosli
Hassan, Osman
Kamaruddin, Kamarulzaman
Abdul Aziz, Suraini
Md. Salleh, Madihah
Wan Md. Zain, Wan Salwanis
author_facet Chee, Chit Lai
Md. Illias, Rosli
Hassan, Osman
Kamaruddin, Kamarulzaman
Abdul Aziz, Suraini
Md. Salleh, Madihah
Wan Md. Zain, Wan Salwanis
author_sort Chee, Chit Lai
building UTeM Institutional Repository
collection Online Access
description Pullulanases (pullulan-6-glucanohydrolase, EC 3.2.1.41) are debranching enzymes that are able to hydrolyze α-1,6-glycosidic linkages in pullulan and branched polysaccharides, producing maltotriose. Pullulanases are a member of family 13 glycosyl hydrolases or α-amylase family. Pullulanase are classified in two categories based on substrate specificity: Type I pullulanases that only hydrolyze α-1,6 linkages, and Type II pullulanases that hydrolyze α-1,6 and α-1,4 linkages. Due to this debranching ability, pullulanases are used in combination with α-glucosidase to improve saccharification rate and yield. In this project, local isolated bacteria were screened for the pullulanase activity by using AZCL-pullulan and Red pullulan. Bacteria S7 and bacteria P2 exhibited the ability to degrade AZCL-pullulan. However, Red pullulan plate growth with bacteria P2 exhibited halo zone. This result confirmed that bacteria P2, which was isolated from local resources, is a pullulanase-producer. I kb pullulanase gene fragment was amplified using PCR. Four conserved regions of amylolytic enzyme and a highly conserved region of pullulanase type I (YNWGYDP) were identified within the deduced amino acid sequence. Bacteria P2 was identified as Exiguohacterium,sp. MAA-I using 16S rRNA gene sequence analysis.
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format Conference or Workshop Item
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institution Universiti Teknologi Malaysia
institution_category Local University
language English
last_indexed 2025-11-15T20:51:34Z
publishDate 2005
recordtype eprints
repository_type Digital Repository
spelling utm-53702017-08-28T08:32:15Z http://eprints.utm.my/5370/ Cloning of pullulanase gene from local isolated bacteria Chee, Chit Lai Md. Illias, Rosli Hassan, Osman Kamaruddin, Kamarulzaman Abdul Aziz, Suraini Md. Salleh, Madihah Wan Md. Zain, Wan Salwanis T Technology (General) Pullulanases (pullulan-6-glucanohydrolase, EC 3.2.1.41) are debranching enzymes that are able to hydrolyze α-1,6-glycosidic linkages in pullulan and branched polysaccharides, producing maltotriose. Pullulanases are a member of family 13 glycosyl hydrolases or α-amylase family. Pullulanase are classified in two categories based on substrate specificity: Type I pullulanases that only hydrolyze α-1,6 linkages, and Type II pullulanases that hydrolyze α-1,6 and α-1,4 linkages. Due to this debranching ability, pullulanases are used in combination with α-glucosidase to improve saccharification rate and yield. In this project, local isolated bacteria were screened for the pullulanase activity by using AZCL-pullulan and Red pullulan. Bacteria S7 and bacteria P2 exhibited the ability to degrade AZCL-pullulan. However, Red pullulan plate growth with bacteria P2 exhibited halo zone. This result confirmed that bacteria P2, which was isolated from local resources, is a pullulanase-producer. I kb pullulanase gene fragment was amplified using PCR. Four conserved regions of amylolytic enzyme and a highly conserved region of pullulanase type I (YNWGYDP) were identified within the deduced amino acid sequence. Bacteria P2 was identified as Exiguohacterium,sp. MAA-I using 16S rRNA gene sequence analysis. 2005 Conference or Workshop Item PeerReviewed application/pdf en http://eprints.utm.my/5370/1/CheeChitLai2005_CloningOfPullulanaseGeneFromLocalIsolatedBacteria.pdf Chee, Chit Lai and Md. Illias, Rosli and Hassan, Osman and Kamaruddin, Kamarulzaman and Abdul Aziz, Suraini and Md. Salleh, Madihah and Wan Md. Zain, Wan Salwanis (2005) Cloning of pullulanase gene from local isolated bacteria. In: 27th Symposium of Malaysian Society for Microbiology, 24-27th November 2005, Grand Plaza Park Royal Penang.
spellingShingle T Technology (General)
Chee, Chit Lai
Md. Illias, Rosli
Hassan, Osman
Kamaruddin, Kamarulzaman
Abdul Aziz, Suraini
Md. Salleh, Madihah
Wan Md. Zain, Wan Salwanis
Cloning of pullulanase gene from local isolated bacteria
title Cloning of pullulanase gene from local isolated bacteria
title_full Cloning of pullulanase gene from local isolated bacteria
title_fullStr Cloning of pullulanase gene from local isolated bacteria
title_full_unstemmed Cloning of pullulanase gene from local isolated bacteria
title_short Cloning of pullulanase gene from local isolated bacteria
title_sort cloning of pullulanase gene from local isolated bacteria
topic T Technology (General)
url http://eprints.utm.my/5370/
http://eprints.utm.my/5370/1/CheeChitLai2005_CloningOfPullulanaseGeneFromLocalIsolatedBacteria.pdf