The use of crude cell extracts of lactic acid bacteria optimized for beta-galactosidase activity to form galactooligosaccharides with lactose, mannose, fucose, and N-acetylglucosamineReport as inadecuate




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Mannose, Beta-galactosidase, N-acetylglucosamine, Oligosaccharides, Galactooligosaccharides, Crude cell extracts, Lactic acid bacteria, Fucose

Lee, Vivian Shin Yuan

Supervisor and department: Gänzle, Michael Agricultural, Food, and Nutritional Sciences Jelen, Paul Agricultural, Food, and Nutritional Sciences

Examining committee member and department: Steiner, Bodo Rural Economy Gänzle, Michael Agricultural, Food, and Nutritional Sciences Jelen, Paul Agricultural, Food, and Nutritional Sciences

Department: Department of Agricultural, Food, and Nutritional Science

Specialization:

Date accepted: 2009-08-27T14:57:12Z

Graduation date: 2009-11

Degree: Master of Science

Degree level: Master's

Abstract: Several lactic acid bacteria contain β-galactosidases. Beta galactosidases catalyze lactose hydrolysis and transfer acceptor sugars onto galactose, producing galactooligosaccharides. The aim of this work was to exploit β-galactosidases of lactic acid bacteria as crude cell extracts to produce novel oligosaccharides with mannose, N-acetylglucosamine, and fucose. Of 17 strains of lactic acid bacteria, transferase activity was the strongest in crude cell extracts of Lactobacillus delbrueckii subsp. bulgaricus, followed by Streptococcus thermophilus, Lactobacillus animalis, and Lactobacillus reuteri in a buffered 19% w-w lactose solution. Incorporation of 6 % w-w glycerol increased transferase activity and enzyme stability at higher incubation temperatures. Incorporation of 10% w-w mannose, N-acetylglucosamine and fucose as acceptor sugars yielded three distinct oligosaccharides with mannose and two with N-acetylglucosamine and fucose, with the composition confirmed using gas chromatography-mass spectrometry. This is the first public report indicating production of oligosaccharides containing N-acetylglucosamine and fucose from β-galactosidases of lactic acid bacteria.

Language: English

DOI: doi:10.7939-R3GX28

Rights: Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of these terms. The author reserves all other publication and other rights in association with the copyright in the thesis and, except as herein before provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the author's prior written permission.





Author: Lee, Vivian Shin Yuan

Source: https://era.library.ualberta.ca/


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University of Alberta The use of crude cell extracts of lactic acid bacteria optimized for betagalactosidase activity to form galactooligosaccharides with lactose, mannose, fucose, and N-acetylglucosamine by Vivian Shin Yuan Lee A thesis submitted to the Faculty of Graduate Studies and Research in partial fulfillment of the requirements for the degree of Master of Sciences Agricultural, Food, and Nutritional Sciences ©Vivian Shin Yuan Lee Fall 2009 Edmonton, Alberta Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only.
Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of these terms. The author reserves all other publication and other rights in association with the copyright in the thesis and, except as herein before provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the authors prior written permission. 1 Examining Committee Michael Gänzle, Agricultural, Food, and Nutritional Sciences Paul Jelen, Agricultural, Food, and Nutritional Sciences Bodo Steiner, Rural Economy, University of Alberta 2 Dedicated to my inspiring colleagues and my supportive family who have encouraged my pursuit of higher education 3 Abstract Several lactic acid bacteria contain β-galactosidases.
Beta galactosidases catalyze lactose hydrolysis and transfer acceptor sugars onto galactose, producing galactooligosaccharides.
The aim of this work was to exploit β-galactosidases of lactic acid bacteria as crude cell extracts to produce novel oligosaccharides with mannose, N-acetylglucosamine, and fucose.
Of 17 strains of lactic acid bacteria, transferase activity was the strongest in crude cell extracts of Lactobacillus delbrueckii subsp.
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