Production and partial purification of lipase produced by Burkholderia cenocepacia ST8 using used engine oil as substrate

Pollution of environment from the impacts of improper disposal of used engine oil (UEO) that lead to the endangering of biota had become a global environmental issue nowadays. UEO contains aliphatic and aromatic hydrocarbon mixtures. The bioconversion of UEO into useful and cost-effective product...

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Main Author:	Lau, Hui Lane
Format:	Thesis
Language:	English English
Published:	2020
Subjects:	Burkholderiales Petroleum as fuel Biodegradation
Online Access:	http://psasir.upm.edu.my/id/eprint/112943/1/112943.pdf
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spelling	my-upm-ir.1129432024-10-22T09:07:24Z Production and partial purification of lipase produced by Burkholderia cenocepacia ST8 using used engine oil as substrate 2020-12 Lau, Hui Lane Pollution of environment from the impacts of improper disposal of used engine oil (UEO) that lead to the endangering of biota had become a global environmental issue nowadays. UEO contains aliphatic and aromatic hydrocarbon mixtures. The bioconversion of UEO into useful and cost-effective product (i.e., lipase) via microbial fermentation, can reduce the risk of releasing UEO into the environment seems to be very attractive strategy. The aim of this study was to investigate the production, optimization, and partial purification of extracellular lipase by an indigenous Burkholderia cenocepacia ST8 in submerged fermentation using shake flask. The effect of lipid and the components of UEO employed for enhancement of lipase production were investigated. Statisticalbased approaches were employed to optimize the fermentation medium based on UEO for the improvement of lipase production by B. cenocepacia. The crude lipase from B. cenocepacia was also partially purified by solvent/salt-based aqueous two-phase systems (ATPS) using UEO-based fermentation broth as feedstock. Preliminary results showed that lipase production by B. cenocepacia using UEO was induced in the presence of Tween 80 in the culture. The role of UEO fractions (hydrocarbon content) in the presence of Tween 80 in relation to the enhancement of lipase production was also studied. On individual hydrocarbon system, aliphatic hydrocarbon was found to be more favorable as compared to aromatic hydrocarbon in the enhancement of lipase production. The use of the mixtures of aliphatic (C15 or C16) and aromatic hydrocarbon gave substantial increment in growth of B. cenocepacia and lipase production as compared to basal medium (BM). Lipase production could be improved by the addition of C40 or benzene, in BM containing UEO. Response surface methodology (RSM) predicted the optimized media constituents to be 2.28% Tween 80 (v/v), 2.26% UEO (v/v), 0.79% nutrient broth (w/v) and 1.33% gum arabic (w/v), with actual observed lipase activity of 216 U/mL. The RSM-optimized system as well as ANN-GA optimized system revealed approximately 1.6-fold increment in lipase production as compared to non-optimized medium. Nutrient broth and Tween 80 were found to be the most important variables that greatly influenced the lipase activity. Partial purification of lipase was obtained with a purification-fold of 4.8 and RT of 141% in a 1-propanol/sodium citrate ATPS system. Enhancement of lipase production is associated with the types of hydrocarbons, individual and in combination as well as the presence of surfactant. The combined effects of aliphatic and aromatic hydrocarbons (UEO content) in the presence of Tween 80 can enhance lipase production. RSM and ANN-GA approaches are effective tools for predicting and optimizing fermentation medium for lipase production. Solvent/salt-based ATPS considered as economically feasible and potential downstream method to purify lipase with high enzyme recovery from UEObased fermentation broth as feedstock. Burkholderiales Petroleum as fuel Biodegradation 2020-12 Thesis http://psasir.upm.edu.my/id/eprint/112943/ http://psasir.upm.edu.my/id/eprint/112943/1/112943.pdf text en public doctoral Universiti Putra Malaysia Burkholderiales Petroleum as fuel Biodegradation Ariff, Arbakariya English
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English English
advisor	Ariff, Arbakariya
topic	Burkholderiales Petroleum as fuel Biodegradation
spellingShingle	Burkholderiales Petroleum as fuel Biodegradation Lau, Hui Lane Production and partial purification of lipase produced by Burkholderia cenocepacia ST8 using used engine oil as substrate
description	Pollution of environment from the impacts of improper disposal of used engine oil (UEO) that lead to the endangering of biota had become a global environmental issue nowadays. UEO contains aliphatic and aromatic hydrocarbon mixtures. The bioconversion of UEO into useful and cost-effective product (i.e., lipase) via microbial fermentation, can reduce the risk of releasing UEO into the environment seems to be very attractive strategy. The aim of this study was to investigate the production, optimization, and partial purification of extracellular lipase by an indigenous Burkholderia cenocepacia ST8 in submerged fermentation using shake flask. The effect of lipid and the components of UEO employed for enhancement of lipase production were investigated. Statisticalbased approaches were employed to optimize the fermentation medium based on UEO for the improvement of lipase production by B. cenocepacia. The crude lipase from B. cenocepacia was also partially purified by solvent/salt-based aqueous two-phase systems (ATPS) using UEO-based fermentation broth as feedstock. Preliminary results showed that lipase production by B. cenocepacia using UEO was induced in the presence of Tween 80 in the culture. The role of UEO fractions (hydrocarbon content) in the presence of Tween 80 in relation to the enhancement of lipase production was also studied. On individual hydrocarbon system, aliphatic hydrocarbon was found to be more favorable as compared to aromatic hydrocarbon in the enhancement of lipase production. The use of the mixtures of aliphatic (C15 or C16) and aromatic hydrocarbon gave substantial increment in growth of B. cenocepacia and lipase production as compared to basal medium (BM). Lipase production could be improved by the addition of C40 or benzene, in BM containing UEO. Response surface methodology (RSM) predicted the optimized media constituents to be 2.28% Tween 80 (v/v), 2.26% UEO (v/v), 0.79% nutrient broth (w/v) and 1.33% gum arabic (w/v), with actual observed lipase activity of 216 U/mL. The RSM-optimized system as well as ANN-GA optimized system revealed approximately 1.6-fold increment in lipase production as compared to non-optimized medium. Nutrient broth and Tween 80 were found to be the most important variables that greatly influenced the lipase activity. Partial purification of lipase was obtained with a purification-fold of 4.8 and RT of 141% in a 1-propanol/sodium citrate ATPS system. Enhancement of lipase production is associated with the types of hydrocarbons, individual and in combination as well as the presence of surfactant. The combined effects of aliphatic and aromatic hydrocarbons (UEO content) in the presence of Tween 80 can enhance lipase production. RSM and ANN-GA approaches are effective tools for predicting and optimizing fermentation medium for lipase production. Solvent/salt-based ATPS considered as economically feasible and potential downstream method to purify lipase with high enzyme recovery from UEObased fermentation broth as feedstock.
format	Thesis
qualification_level	Doctorate
author	Lau, Hui Lane
author_facet	Lau, Hui Lane
author_sort	Lau, Hui Lane
title	Production and partial purification of lipase produced by Burkholderia cenocepacia ST8 using used engine oil as substrate
title_short	Production and partial purification of lipase produced by Burkholderia cenocepacia ST8 using used engine oil as substrate
title_full	Production and partial purification of lipase produced by Burkholderia cenocepacia ST8 using used engine oil as substrate
title_fullStr	Production and partial purification of lipase produced by Burkholderia cenocepacia ST8 using used engine oil as substrate
title_full_unstemmed	Production and partial purification of lipase produced by Burkholderia cenocepacia ST8 using used engine oil as substrate
title_sort	production and partial purification of lipase produced by burkholderia cenocepacia st8 using used engine oil as substrate
granting_institution	Universiti Putra Malaysia
publishDate	2020
url	http://psasir.upm.edu.my/id/eprint/112943/1/112943.pdf
_version_	1818586128465264640

Production and partial purification of lipase produced by Burkholderia cenocepacia ST8 using used engine oil as substrate

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