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Title: Evaluation of β-galactosidase from Lactobacillus acidophilus as biocatalyst for galacto-oligosaccharides synthesis: Product structural characterization and enzyme immobilization

Abstract

β-Galactosidase is an important industrial enzyme that catalyzes reaction of lactose hydrolysis and recently more interesting reaction of transgalactosylation, yielding a highly valuable group of prebiotic compounds named galacto-oligosaccharides (GOS). In this paper, parameters for achieving high yields of tailor-made GOS using crude β-galactosidase obtained from Lactobacillus acidophilus ATCC 4356, probiotic bacteria regarded as safe for human consumption, were optimized. At the same time, detailed structural elucidation of obtained GOS was conducted, and it was concluded that β-galactosidase from L. acidophilus shows a particular specificity towards the formation of β-(1 → 6) glycosidic bonds. In order to develop more stable and economically cost-effective preparation, crude enzyme was successfully immobilized on a methacrylic polymer carrier Lifetech ECR8409, leading to its simultaneous 2-fold purification. In conclusion, this immobilized preparation showed unchanged specificity towards the transgalactosylation reaction, thus yielding 86 g/l GOS under the previously optimized conditions (lactose concentration 400 g/l in 0.1 M sodium phosphate buffer, pH 6.8 and temperature 50°C).

Authors:
 [1];  [1];  [1];  [2];  [2];  [3];  [1]
  1. Univ. of Belgrade, Belgrade (Serbia)
  2. INRA, Nantes (France)
  3. Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1459008
Grant/Contract Number:  
46010; AC05-76RL01830
Resource Type:
Accepted Manuscript
Journal Name:
Journal of Bioscience and Bioengineering
Additional Journal Information:
Journal Volume: 126; Journal Issue: 6; Journal ID: ISSN 1389-1723
Publisher:
Society for Bioscience and Bioengineering, c/o Osaka University., Faculty of Engineering, Yamadaoka 2-1, Suita-shi, Osaka Japan
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; β-Galactosidase; Immobilization; Lactobacillus acidophilus; Galacto-oligosaccharides; Lifetech ECR resins

Citation Formats

Carevic, Milica, Vukasinovic-Sekulic, Maja, Corovic, Marija, Rogniaux, Helene, Ropartz, David, Velickovic, Dusan, and Bezbradica, Dejan. Evaluation of β-galactosidase from Lactobacillus acidophilus as biocatalyst for galacto-oligosaccharides synthesis: Product structural characterization and enzyme immobilization. United States: N. p., 2018. Web. https://doi.org/10.1016/j.jbiosc.2018.06.003.
Carevic, Milica, Vukasinovic-Sekulic, Maja, Corovic, Marija, Rogniaux, Helene, Ropartz, David, Velickovic, Dusan, & Bezbradica, Dejan. Evaluation of β-galactosidase from Lactobacillus acidophilus as biocatalyst for galacto-oligosaccharides synthesis: Product structural characterization and enzyme immobilization. United States. https://doi.org/10.1016/j.jbiosc.2018.06.003
Carevic, Milica, Vukasinovic-Sekulic, Maja, Corovic, Marija, Rogniaux, Helene, Ropartz, David, Velickovic, Dusan, and Bezbradica, Dejan. Sat . "Evaluation of β-galactosidase from Lactobacillus acidophilus as biocatalyst for galacto-oligosaccharides synthesis: Product structural characterization and enzyme immobilization". United States. https://doi.org/10.1016/j.jbiosc.2018.06.003. https://www.osti.gov/servlets/purl/1459008.
@article{osti_1459008,
title = {Evaluation of β-galactosidase from Lactobacillus acidophilus as biocatalyst for galacto-oligosaccharides synthesis: Product structural characterization and enzyme immobilization},
author = {Carevic, Milica and Vukasinovic-Sekulic, Maja and Corovic, Marija and Rogniaux, Helene and Ropartz, David and Velickovic, Dusan and Bezbradica, Dejan},
abstractNote = {β-Galactosidase is an important industrial enzyme that catalyzes reaction of lactose hydrolysis and recently more interesting reaction of transgalactosylation, yielding a highly valuable group of prebiotic compounds named galacto-oligosaccharides (GOS). In this paper, parameters for achieving high yields of tailor-made GOS using crude β-galactosidase obtained from Lactobacillus acidophilus ATCC 4356, probiotic bacteria regarded as safe for human consumption, were optimized. At the same time, detailed structural elucidation of obtained GOS was conducted, and it was concluded that β-galactosidase from L. acidophilus shows a particular specificity towards the formation of β-(1 → 6) glycosidic bonds. In order to develop more stable and economically cost-effective preparation, crude enzyme was successfully immobilized on a methacrylic polymer carrier Lifetech ECR8409, leading to its simultaneous 2-fold purification. In conclusion, this immobilized preparation showed unchanged specificity towards the transgalactosylation reaction, thus yielding 86 g/l GOS under the previously optimized conditions (lactose concentration 400 g/l in 0.1 M sodium phosphate buffer, pH 6.8 and temperature 50°C).},
doi = {10.1016/j.jbiosc.2018.06.003},
journal = {Journal of Bioscience and Bioengineering},
number = 6,
volume = 126,
place = {United States},
year = {2018},
month = {6}
}

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