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Cloning of pullulanase gene from local isolated bacteria

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.



Pullulanases (pullulan-6-glucanohydrolase, EC 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.

Item Type:Conference or Workshop Item (Poster)
Uncontrolled Keywords:Bacteria, cloning, enzymes, pullulanase gene, saccharification
Subjects:T Technology > T Technology (General)
Divisions:Chemical and Natural Resources Engineering
ID Code:5370
Deposited By: Norhani Jusoh
Deposited On:31 Mar 2008 08:55
Last Modified:28 Aug 2017 08:32

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