Universal Journal of Microbiology Research 7(1): 1-6, 2019 http://www.hrpub.org
DOI: 10.13189/ujmr.2019.070101
Production and Characterization of Lipase Enzyme
Expressed by Crude Oil Contaminated Soil Isolates
Ndubuisi Gabriel Elemuo
1
, Sunday Sunday Ikiensikimama
2
, Chikwendu Ebenezer Ubani
2
,
Evans Chidi Egwim
3
, Justice Obinna Osuoha
1,*
1
World Bank Africa Center of Excellence in Oilfield Chemicals Research, University of Port Harcourt, Nigeria
2
Department of Petroleum and Gas Engineering, University of Port Harcourt, Nigeria
3
Department of Biochemistry, Federal University of Technology, Minna, Nigeria
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Abstract Lipases are hydrolytic enzymes responsible
for the hydrolysis of triacylglycerol into glycerol and free
fatty acids. The biodiversity of crude oil contaminated soil
was explored for the isolation of novel potent lipase
producing microorganisms. In this current study,
microorganisms isolated from crude oil contaminated soils
were screened for lipase activity and expression. Six fungal
strains namely: Yeast, Aspergillus flavus, Aspergillus niger,
Verticillus sp., Penicillum sp., and Microsporum audouini
demonstrated lipase producing potentials and the best two:
Verticillus sp. and Penicillum sp. were selected for enzyme
production and characterization. The lipase enzyme was
produced in broth medium. Optimization of selected
biochemical parameters reveal that optimum temperature
for lipase activity was 40 ºC, pH 7 with an incubation time
of 168 h for lipase expressed by Verticillus sp. and 40 ºC,
pH 8 with an incubation time of 144 h was established for
lipase expressed by Penicillium sp. The kinetic parameters
of the lipase enzyme revealed that the enzymes had low
Km (0.19 and 0.30 respectively) for the selected substrate.
Keywords Lipase, Lipase Production, Kinetic
Parameters, Optimum PH and Temperature
1. Introduction
Lipases, classified as EC 3.1.1.3 are enzymes that
catalyze the complete breakdown of oils and fats by
converting them into glycerol and fatty acids (Jaiswal et
al., 2017). The principal substrate for lipase enzymes are
triacylglycerols. The capacity of lipase to carry out
numerous chemical biotransformation is the basis for their
increasing popularity in the chemical, detergent,
pharmaceutical, food and oil industries. In this regard,
they are extensively utilized for lipolysis of cream and fat,
ripening of cheese, for the enzymatic hydrolysis of milk
and for the enhancement of flavor etc. in the diary and
food industries (Franken et al., 2009). Lipases are also
utilized to augment the absorbency of fabrics in the textile
industry, as additives for the production of washing
powder in the detergent industries, for diverse
transesterification reactions and production of polymers
that are biodegradable (Hasan et al., 2006; Jaiswal et al.,
2017).
In the oil and gas industry, the use of enzymes has been
recently suggested for several applications. A robust of
enzymes has been provided for tolerating oil reservoir
condition by the modern biotechnology industry. Some
applications of enzymes in the oil and gas company were
reported by Harris and Mckay (1998). Such application
includes; breaking of gel in drilling operation to avoid filter
cake formation, removal of Sulphur in hydrocarbon,
production of “enzyme-based acid” which is used for the
purpose of treating formation damage and matrix-acidizing
of carbonate, pre-treatment of biopolymers using enzyme
to improve the handling characteristics of biopolymer.
Furthermore, lipases have also been employed as
catalyst for enhanced production of diverse products
utilized in paper, pulp and cosmetic industry (Jaiswal et al.,
2017). Many thermostable bacterial lipases have been
isolated and characterized (Kumar et al., 2005) from
different ecological zones. Among the identified bacteria,
Pseudomonas sp., Chromobacterium sp., Achromobacter
sp., Serratia sp. and Alcaligenes sp. have been evaluated
for possible production of lipases (Jaiswal et al., 2017).
As a matter of fact, microorganisms that has the capacity
to express lipase can be isolated from numerous habitats
which include: crude oil contaminated soils, deteriorated
food, diary product industries and wastes from vegetable
oil industries to mention but a few (Sharma et al., 2001).
According to Koet et al. (2005), soils entertain diverse
microorganisms that could be potent producers of lipase
and other enzymes of industrial value. The screening of
novel microorganisms isolated from distinct ecological