Production and Characterisation of Cyclodextrin Glycosyltransferase from a Locally Isolated Bacillus Sp

Cyclodextrin glycosyltransferase (E.C.2.4.1.19) synthesise cyclic oligosaccharide which is also known as cyclodextrin, from starch. Most of the known CGTases produce a mixture of a-, B- and Y-CD at different ratios. CGTase producing microorganism was isolated from local soils on selective agar mediu...

Full description

Saved in:

Bibliographic Details
Main Author:	Ai Noi, Sauvaphap
Format:	Thesis
Language:	English English
Published:	2006
Subjects:	Cyclodextrins Glycosyltransferases
Online Access:	http://psasir.upm.edu.my/id/eprint/4843/1/FBSB_2006_35a.pdf
Tags:	Add Tag No Tags, Be the first to tag this record!

id	my-upm-ir.4843
record_format	uketd_dc
spelling	my-upm-ir.48432013-05-27T07:18:37Z Production and Characterisation of Cyclodextrin Glycosyltransferase from a Locally Isolated Bacillus Sp 2006 Ai Noi, Sauvaphap Cyclodextrin glycosyltransferase (E.C.2.4.1.19) synthesise cyclic oligosaccharide which is also known as cyclodextrin, from starch. Most of the known CGTases produce a mixture of a-, B- and Y-CD at different ratios. CGTase producing microorganism was isolated from local soils on selective agar medium containing soluble starch which produced clear zones as qualitative measurement of the enzyme present. A total of 250 isolates were collected but only one isolate (Strain MK 6) was selected for further studies based on its highest activity. Strain MK 6 was identified as gram positive rod, motile and produced spore. Biochemical identification using API CHB/E medium confirmed the strain MK 6 was the Bacilllus sp with 85% similarities. CGTase isolated from alkalophilic Bacillus sp. was further characterized. Optimum activity obtained at temperature of 70oC and the enzyme shows a wide range of pH stability ranging from 4 -10 when stored at 4oC for 24 hours and temperature stability ranging from 30oC - 80oC at 1 h incubation period. The CGTase activity was even maintained at 0.4 U/ml at 90oC for 40 min incubation. Prior to optimisation of CGTase production, selection for the best carbon source through detection on modified phenolphthalein method containing different types of starch were performed. Sago starch gave significant result and was used for further optimisation using statistical analysis namely Response Surface Methodology (RSM). The optimal calculated values were 3.34% sago starch, initial pH of 10.15 and agitation speed of 187 rpm; with predicted activity of 2.07 U/ml of CGTase. These predicted optimal parameters were confirmed in the laboratory and the final CGTase activity obtained was very close to the predicted value at 2.56 U/ml. The optimised crude enzyme produced mainly y-CD (61.6% of the total cyclodextrin amount) with only y-CD as minimal product without detection of y-CD. Cyclodextrins Glycosyltransferases 2006 Thesis http://psasir.upm.edu.my/id/eprint/4843/ http://psasir.upm.edu.my/id/eprint/4843/1/FBSB_2006_35a.pdf application/pdf en public masters Universiti Putra Malaysia Cyclodextrins Glycosyltransferases Faculty of Biotechnology and Biomolecular Sciences English
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English English
topic	Cyclodextrins Glycosyltransferases
spellingShingle	Cyclodextrins Glycosyltransferases Ai Noi, Sauvaphap Production and Characterisation of Cyclodextrin Glycosyltransferase from a Locally Isolated Bacillus Sp
description	Cyclodextrin glycosyltransferase (E.C.2.4.1.19) synthesise cyclic oligosaccharide which is also known as cyclodextrin, from starch. Most of the known CGTases produce a mixture of a-, B- and Y-CD at different ratios. CGTase producing microorganism was isolated from local soils on selective agar medium containing soluble starch which produced clear zones as qualitative measurement of the enzyme present. A total of 250 isolates were collected but only one isolate (Strain MK 6) was selected for further studies based on its highest activity. Strain MK 6 was identified as gram positive rod, motile and produced spore. Biochemical identification using API CHB/E medium confirmed the strain MK 6 was the Bacilllus sp with 85% similarities. CGTase isolated from alkalophilic Bacillus sp. was further characterized. Optimum activity obtained at temperature of 70oC and the enzyme shows a wide range of pH stability ranging from 4 -10 when stored at 4oC for 24 hours and temperature stability ranging from 30oC - 80oC at 1 h incubation period. The CGTase activity was even maintained at 0.4 U/ml at 90oC for 40 min incubation. Prior to optimisation of CGTase production, selection for the best carbon source through detection on modified phenolphthalein method containing different types of starch were performed. Sago starch gave significant result and was used for further optimisation using statistical analysis namely Response Surface Methodology (RSM). The optimal calculated values were 3.34% sago starch, initial pH of 10.15 and agitation speed of 187 rpm; with predicted activity of 2.07 U/ml of CGTase. These predicted optimal parameters were confirmed in the laboratory and the final CGTase activity obtained was very close to the predicted value at 2.56 U/ml. The optimised crude enzyme produced mainly y-CD (61.6% of the total cyclodextrin amount) with only y-CD as minimal product without detection of y-CD.
format	Thesis
qualification_level	Master's degree
author	Ai Noi, Sauvaphap
author_facet	Ai Noi, Sauvaphap
author_sort	Ai Noi, Sauvaphap
title	Production and Characterisation of Cyclodextrin Glycosyltransferase from a Locally Isolated Bacillus Sp
title_short	Production and Characterisation of Cyclodextrin Glycosyltransferase from a Locally Isolated Bacillus Sp
title_full	Production and Characterisation of Cyclodextrin Glycosyltransferase from a Locally Isolated Bacillus Sp
title_fullStr	Production and Characterisation of Cyclodextrin Glycosyltransferase from a Locally Isolated Bacillus Sp
title_full_unstemmed	Production and Characterisation of Cyclodextrin Glycosyltransferase from a Locally Isolated Bacillus Sp
title_sort	production and characterisation of cyclodextrin glycosyltransferase from a locally isolated bacillus sp
granting_institution	Universiti Putra Malaysia
granting_department	Faculty of Biotechnology and Biomolecular Sciences
publishDate	2006
url	http://psasir.upm.edu.my/id/eprint/4843/1/FBSB_2006_35a.pdf
_version_	1747810295690035200

Production and Characterisation of Cyclodextrin Glycosyltransferase from a Locally Isolated Bacillus Sp

Similar Items