Screening And Characterisation Of Potential Lipase Producing Strains From Arctic Samples
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Date
2018-01
Authors
Ramle, Zakiah
Journal Title
Journal ISSN
Volume Title
Publisher
Universiti Sains Malaysia
Abstract
Psychrotolerant microorganisms are widespread in natural cold environments
ranging from deep sea to polar region and have a synergistic mechanism that allows
them to adapt living in the cold. Possessing evolved cold active enzymes is one of the
basic strategies of adaptation. Among the various cold enzymes is cold active microbial
lipase, an enzyme that is able to hydrolyse triglycerols releasing fatty acids. Microbial
lipases have gained notable attention for their widespread uses in industries. However,
bulk production of enzymes direct from original microbe source is not feasible due to
high maintenance cost thus will not be able to cater to high industrial demands.
Developments for efficient lipase productions are still necessary for lipase biocatalysis
to rise to its full potential. This study was conducted to screen and identify suitable
psychrotolerant lipase producing strain to be expressed in a heterologous host expression
system before characterising the recombinant protein. With that in mind, six
microorganisms labelled as ARB 1A, ARB 1B, ARB 1C, ARB 10, ARC 8A and ARC
8B’s DNAs were extracted via modified CTAB method and applied to 16S rDNA
sequence analysis. Gram staining revealed five out of six isolates were Gram negative.
They were identified as bacteria belonging to four genera Pseudomonas sp.,
Arthrobacter sp., Acinetobacter sp., and Janthinobacterium sp., and were screened for
lipase activity. Lipase activity was detected using sensitive and specific plate assay using
olive oil as substrate. ARB 1B, ARB 10 and ARC 8A were tested as lipase positive.
ARB 10, having 97% similarity to Pseudomonas sp. was selected for cloning. Lipase
gene was amplified by PCR method using specific set of primers that was designed to
target the gene of interest.
Description
Keywords
Screening and characterisation of potential lipase , producing strains from arctic samples