Nurhasanah, Nurhasanah and Nurbaiti, Santi and Madayanti, Fida and Akhmaloka, Akhmaloka (2014) INTERNAL PRIMER DESIGN FOR DIVERSITY STUDY OF THERMOSTABLE LIPASE GENE FRAGMENT FROM ENVIRONMENTAL SAMPLE. Prosiding Seminar Nasional Biokimia 2014. pp. 1-5. ISSN ISBN 978-602-98506-1-1

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Lipase is a hydrolytic enzyme that has an important role in the biotechnology industries. This is due to its ability to catalyze multiple organic reactions such as hydrolysis, esterification, interesterification, transesterification and synthesis reactions. Research for inventing novel lipase with unique properties i.e. thermostable lipase is still being carried out by using several approaches. One of the method is by metagenomic approach which is directly amplifying lipase gene from natural resources. Hence the aim of this research is to design a primers to amplify lipase gene fragments from environmental samples. The study was conducted through bioinformatics analysis of lipase database from GeneBank, selection of conserved region of the whole lipase genes, primers and in silico analysis by using Clustal X, Gendoc, Primer Detective and GenMon softwares. 68 amino acid sequences from 68 bacteria genus were used to obtain the conserved region of lipase gene. The results showed that there are 4 conserved regions, namely one, second, third and fourth region. The best primer pair was obtained from second and fourth regions, and resulted lipase gene fragments for approximately 570 bp. Based on in silico study indicated that the primers amplify some group of bacteria. Therefore, we conclude the primer can be used to amplify lipase gene fragments from environmental sample in the form of a mixture of lipase gene fragments from various bacteria. Several lipase fragments were detected on polyacrylamide gel after separation of PCR mixtures by using DGGE apparatus.

Item Type: Article
Subjects: Q Science > QD Chemistry
Divisions: Fakultas Matematika dan Ilmu Pengetahuan Alam (FMIPA) > Prodi Kimia
Depositing User: Nurhasanah Nurhasanah Husin
Date Deposited: 29 Dec 2016 01:06
Last Modified: 29 Dec 2016 01:06

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