Using pcr to identify bacterial strains owning high lipase activity based on their single nucleotide polymorphisms

Nguyen Quynh Uyen, Nguyen Ha Xuyen, Phan Thi Ha, Nguyen Huynh Minh Quyen, Vo Thi Thuong Lan


Lipases (triacylglycerol acyl hydrolases, E.C. are a class of enzyme that can catalyze lipolytic, acidolysis, esterification or trans-esterification under different conditions. Lipases, especially from microbial resources, have been of great interest due to their applications in many fields. However, so far the titrimetric methods, such as the methods using olive oil, p-nitrophenylpalimitate or phenyl acetate as a substrate, are too expensive and/or time consuming. In this study, a convenient method that can identify many samples with high lipase activity was established. In particular, six fragments of the strains with different patterns of single-strand conformation polymorphism SSCP were observed among 20 bacterial strains owing lipase activity. These fragments were cloned into pGEM-T Easy vector in order to sequence their nucleotides and then identify their SNPs. The PCR primer set was designed based on the SNPs of the lipase sequences and afterward this newly designed primer and published one were used to screen the 20 strains via PCR. The primary result showed that 15/20 strains gave a positive result of PCR product and 11 out of these 15 strains (73.3%) had high lipase activity (more than 3.25 Unit/ml).


Lipase activity, single strain conformation polymorphism (SSCP), single nucleotide polymorphism (SNP), PCR

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