Purification of fusion peptide biosurfactant using combined chemical extraction and cross-flow microfiltration

Biosurfactant becomes important nowadays as sustainable bio-derived surfactants emerged to replace the petroleum based surfactant. In this work, the fusion peptide biosurfactant (HSG) was successfully expressed when induced with 1 mM Isopropyl-ß-D-thiogalactopyranoside (IPTG). The combined process o...

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Main Author:	Leow, Chee Who
Format:	Thesis
Language:	English
Published:	2011
Subjects:	TP Chemical technology
Online Access:	http://eprints.utm.my/id/eprint/32782/1/LeowCheeWohMFKA2011.pdf
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spelling	my-utm-ep.327822018-05-27T07:51:40Z Purification of fusion peptide biosurfactant using combined chemical extraction and cross-flow microfiltration 2011-07 Leow, Chee Who TP Chemical technology Biosurfactant becomes important nowadays as sustainable bio-derived surfactants emerged to replace the petroleum based surfactant. In this work, the fusion peptide biosurfactant (HSG) was successfully expressed when induced with 1 mM Isopropyl-ß-D-thiogalactopyranoside (IPTG). The combined process of chemical extraction and cross-flow microfiltration was conducted to disrupt the cell membrane and isolate the desired product, HSG. A combination of different concentration of Triton X-100 and Ethylenediaminetetraacetic Acid (EDTA) was investigated to obtain maximized protein extraction from the cells. The efficiency of the chemical extraction was compared with B-PER commercial bacterial extraction kit. Protein estimation was performed using Bicinchoninic Acid (BCA) Protein Assay method and Polyacrylamide Gel Electrophoresis (PAGE) analysis. The results showed that the combination of 1% vv-1 Triton X-100 and 1 mM EDTA released the highest amount of soluble protein and is comparable to the B-PER commercial bacterial extraction kit. The extraction broth is then applied to cross-flow microfiltration process with a 0.2 µm polysulfone hollow fiber membrane. The effect of rotor speed and transmembrane pressure (TMP) for peptide transmission were investigated. The rotor speed for the microfiltration test was varied at 150, 200 and 300 rpm which gave rise to the uncontrolled TMP of 2.5, 2.5 and 4 psig respectively. The highest overall permeate flux achieved at 300 rpm was selected for further investigation at two different TMP of 4 and 5 psig. It was found that the operating conditions at 300 rpm and 5 psig gave 36.33% more protein transmission as compared to operation at 300 rpm and 4 psig. Backpulsing was applied to the microfiltration system to minimize the fouling problem. An overall protein transmission of about 59.6% was achieved with the operating parameter of 300 rpm at constant transmembrane pressure of 5 psig. 2011-07 Thesis http://eprints.utm.my/id/eprint/32782/ http://eprints.utm.my/id/eprint/32782/1/LeowCheeWohMFKA2011.pdf application/pdf en public masters Universiti Teknologi Malaysia, Faculty of Chemical Engineering Faculty of Chemical Engineering
institution	Universiti Teknologi Malaysia
collection	UTM Institutional Repository
language	English
topic	TP Chemical technology
spellingShingle	TP Chemical technology Leow, Chee Who Purification of fusion peptide biosurfactant using combined chemical extraction and cross-flow microfiltration
description	Biosurfactant becomes important nowadays as sustainable bio-derived surfactants emerged to replace the petroleum based surfactant. In this work, the fusion peptide biosurfactant (HSG) was successfully expressed when induced with 1 mM Isopropyl-ß-D-thiogalactopyranoside (IPTG). The combined process of chemical extraction and cross-flow microfiltration was conducted to disrupt the cell membrane and isolate the desired product, HSG. A combination of different concentration of Triton X-100 and Ethylenediaminetetraacetic Acid (EDTA) was investigated to obtain maximized protein extraction from the cells. The efficiency of the chemical extraction was compared with B-PER commercial bacterial extraction kit. Protein estimation was performed using Bicinchoninic Acid (BCA) Protein Assay method and Polyacrylamide Gel Electrophoresis (PAGE) analysis. The results showed that the combination of 1% vv-1 Triton X-100 and 1 mM EDTA released the highest amount of soluble protein and is comparable to the B-PER commercial bacterial extraction kit. The extraction broth is then applied to cross-flow microfiltration process with a 0.2 µm polysulfone hollow fiber membrane. The effect of rotor speed and transmembrane pressure (TMP) for peptide transmission were investigated. The rotor speed for the microfiltration test was varied at 150, 200 and 300 rpm which gave rise to the uncontrolled TMP of 2.5, 2.5 and 4 psig respectively. The highest overall permeate flux achieved at 300 rpm was selected for further investigation at two different TMP of 4 and 5 psig. It was found that the operating conditions at 300 rpm and 5 psig gave 36.33% more protein transmission as compared to operation at 300 rpm and 4 psig. Backpulsing was applied to the microfiltration system to minimize the fouling problem. An overall protein transmission of about 59.6% was achieved with the operating parameter of 300 rpm at constant transmembrane pressure of 5 psig.
format	Thesis
qualification_level	Master's degree
author	Leow, Chee Who
author_facet	Leow, Chee Who
author_sort	Leow, Chee Who
title	Purification of fusion peptide biosurfactant using combined chemical extraction and cross-flow microfiltration
title_short	Purification of fusion peptide biosurfactant using combined chemical extraction and cross-flow microfiltration
title_full	Purification of fusion peptide biosurfactant using combined chemical extraction and cross-flow microfiltration
title_fullStr	Purification of fusion peptide biosurfactant using combined chemical extraction and cross-flow microfiltration
title_full_unstemmed	Purification of fusion peptide biosurfactant using combined chemical extraction and cross-flow microfiltration
title_sort	purification of fusion peptide biosurfactant using combined chemical extraction and cross-flow microfiltration
granting_institution	Universiti Teknologi Malaysia, Faculty of Chemical Engineering
granting_department	Faculty of Chemical Engineering
publishDate	2011
url	http://eprints.utm.my/id/eprint/32782/1/LeowCheeWohMFKA2011.pdf
_version_	1747816059157610496

Purification of fusion peptide biosurfactant using combined chemical extraction and cross-flow microfiltration

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