Optimisation of enzymatic reactions for galactooligosaccharides synthesis using beta-galactanase from Geobacillus mahadii Geo-05

Galacto-oligosaccharide (GOS) is one of the prominent prebiotics used in infant formula milk. Endo-beta-1,4-galactanase hydrolyses galactan and produces GOS with different degree of polymerisation (DP). DP of the GOS could vary depending on the enzyme source. Besides, reaction parameters such as enz...

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Main Author:	Ismail, Noor Faizah
Format:	Thesis
Language:	English
Published:	2022
Subjects:	TP Chemical technology
Online Access:	http://eprints.utm.my/id/eprint/101590/1/NoorFaizahIsmailPChe2022.pdf
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spelling	my-utm-ep.1015902023-06-26T06:50:37Z Optimisation of enzymatic reactions for galactooligosaccharides synthesis using beta-galactanase from Geobacillus mahadii Geo-05 2022 Ismail, Noor Faizah TP Chemical technology Galacto-oligosaccharide (GOS) is one of the prominent prebiotics used in infant formula milk. Endo-beta-1,4-galactanase hydrolyses galactan and produces GOS with different degree of polymerisation (DP). DP of the GOS could vary depending on the enzyme source. Besides, reaction parameters such as enzyme loading, substrate loading, incubation temperature, pH and reaction time also contribute to various DP of GOS synthesis. In this study, beta-galactanase from Geobacillus mahadii Geo-05 was synthesised with a codon optimised sequence (named BgcGC) and biochemically characterised for its optimum function. The BgcGC reaction was optimised to obtain the highest yield of the targeted GOS (galactotriose, G3 and galactotetraose, G4) from the potato galactan. Then, the optimised conditions were applied on pectin extracted from sweet potato peels (SPP) to produce the GOS. BgcGC could actively hydrolyse potato galactan at 60°C in McIlvaine buffer pH 6 and stable to a wide range of pH between 4 to 10. According to the screening by the factorial design, three reaction parameters significantly contributed to the highest yield of GOS. Due to curvature prediction, the central composite design was further applied to optimise BgcGC reaction to produce GOS. The optimum reaction parameters for the GOS synthesis were identified at 0.15 U/ml BgcGC loading, 1.5 h incubation time and 50°C reaction temperature yielding a 1.5-fold increment of the GOS synthesis. The reaction of BgcGC towards SPP pectin resulted in a 0.081 mg/g yield of G3. These findings showed that GOS synthesised by BgcGC could be enhanced through the manipulation of reaction parameters with a notable increment of GOS yield. Moreover, BgcGC exhibited the potential to be used as an enzyme cocktail for the depolymerisation of pectin polysaccharides. 2022 Thesis http://eprints.utm.my/id/eprint/101590/ http://eprints.utm.my/id/eprint/101590/1/NoorFaizahIsmailPChe2022.pdf application/pdf en public http://dms.library.utm.my:8080/vital/access/manager/Repository/vital:150697 phd doctoral Universiti Teknologi Malaysia Faculty of Engineering - School of Chemical & Energy Engineering
institution	Universiti Teknologi Malaysia
collection	UTM Institutional Repository
language	English
topic	TP Chemical technology
spellingShingle	TP Chemical technology Ismail, Noor Faizah Optimisation of enzymatic reactions for galactooligosaccharides synthesis using beta-galactanase from Geobacillus mahadii Geo-05
description	Galacto-oligosaccharide (GOS) is one of the prominent prebiotics used in infant formula milk. Endo-beta-1,4-galactanase hydrolyses galactan and produces GOS with different degree of polymerisation (DP). DP of the GOS could vary depending on the enzyme source. Besides, reaction parameters such as enzyme loading, substrate loading, incubation temperature, pH and reaction time also contribute to various DP of GOS synthesis. In this study, beta-galactanase from Geobacillus mahadii Geo-05 was synthesised with a codon optimised sequence (named BgcGC) and biochemically characterised for its optimum function. The BgcGC reaction was optimised to obtain the highest yield of the targeted GOS (galactotriose, G3 and galactotetraose, G4) from the potato galactan. Then, the optimised conditions were applied on pectin extracted from sweet potato peels (SPP) to produce the GOS. BgcGC could actively hydrolyse potato galactan at 60°C in McIlvaine buffer pH 6 and stable to a wide range of pH between 4 to 10. According to the screening by the factorial design, three reaction parameters significantly contributed to the highest yield of GOS. Due to curvature prediction, the central composite design was further applied to optimise BgcGC reaction to produce GOS. The optimum reaction parameters for the GOS synthesis were identified at 0.15 U/ml BgcGC loading, 1.5 h incubation time and 50°C reaction temperature yielding a 1.5-fold increment of the GOS synthesis. The reaction of BgcGC towards SPP pectin resulted in a 0.081 mg/g yield of G3. These findings showed that GOS synthesised by BgcGC could be enhanced through the manipulation of reaction parameters with a notable increment of GOS yield. Moreover, BgcGC exhibited the potential to be used as an enzyme cocktail for the depolymerisation of pectin polysaccharides.
format	Thesis
qualification_name	Doctor of Philosophy (PhD.)
qualification_level	Doctorate
author	Ismail, Noor Faizah
author_facet	Ismail, Noor Faizah
author_sort	Ismail, Noor Faizah
title	Optimisation of enzymatic reactions for galactooligosaccharides synthesis using beta-galactanase from Geobacillus mahadii Geo-05
title_short	Optimisation of enzymatic reactions for galactooligosaccharides synthesis using beta-galactanase from Geobacillus mahadii Geo-05
title_full	Optimisation of enzymatic reactions for galactooligosaccharides synthesis using beta-galactanase from Geobacillus mahadii Geo-05
title_fullStr	Optimisation of enzymatic reactions for galactooligosaccharides synthesis using beta-galactanase from Geobacillus mahadii Geo-05
title_full_unstemmed	Optimisation of enzymatic reactions for galactooligosaccharides synthesis using beta-galactanase from Geobacillus mahadii Geo-05
title_sort	optimisation of enzymatic reactions for galactooligosaccharides synthesis using beta-galactanase from geobacillus mahadii geo-05
granting_institution	Universiti Teknologi Malaysia
granting_department	Faculty of Engineering - School of Chemical & Energy Engineering
publishDate	2022
url	http://eprints.utm.my/id/eprint/101590/1/NoorFaizahIsmailPChe2022.pdf
_version_	1776100734047092736

Optimisation of enzymatic reactions for galactooligosaccharides synthesis using beta-galactanase from Geobacillus mahadii Geo-05

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