Publication

Biochemical characterization of the functional roles of residues in the active site of the β-galactosidase from Bacillus circulans ATCC 31382

Yin, H., Pijning, T., Meng, X., Dijkhuizen, L. & van Leeuwen, S. S., 2017, In : Biochemistry. 56, 24, p. 3109-3118 9 p.

Research output: Contribution to journalArticleAcademicpeer-review

APA

Yin, H., Pijning, T., Meng, X., Dijkhuizen, L., & van Leeuwen, S. S. (2017). Biochemical characterization of the functional roles of residues in the active site of the β-galactosidase from Bacillus circulans ATCC 31382. Biochemistry, 56(24), 3109-3118. https://doi.org/10.1021/acs.biochem.7b00207

Author

Yin, Huifang ; Pijning, Tjaard ; Meng, Xiangfeng ; Dijkhuizen, Lubbert ; van Leeuwen, Sander Sebastiaan. / Biochemical characterization of the functional roles of residues in the active site of the β-galactosidase from Bacillus circulans ATCC 31382. In: Biochemistry. 2017 ; Vol. 56, No. 24. pp. 3109-3118.

Harvard

Yin, H, Pijning, T, Meng, X, Dijkhuizen, L & van Leeuwen, SS 2017, 'Biochemical characterization of the functional roles of residues in the active site of the β-galactosidase from Bacillus circulans ATCC 31382', Biochemistry, vol. 56, no. 24, pp. 3109-3118. https://doi.org/10.1021/acs.biochem.7b00207

Standard

Biochemical characterization of the functional roles of residues in the active site of the β-galactosidase from Bacillus circulans ATCC 31382. / Yin, Huifang; Pijning, Tjaard; Meng, Xiangfeng; Dijkhuizen, Lubbert; van Leeuwen, Sander Sebastiaan.

In: Biochemistry, Vol. 56, No. 24, 2017, p. 3109-3118.

Research output: Contribution to journalArticleAcademicpeer-review

Vancouver

Yin H, Pijning T, Meng X, Dijkhuizen L, van Leeuwen SS. Biochemical characterization of the functional roles of residues in the active site of the β-galactosidase from Bacillus circulans ATCC 31382. Biochemistry. 2017;56(24):3109-3118. https://doi.org/10.1021/acs.biochem.7b00207


BibTeX

@article{fb811518ceef4b629f01387a12232c18,
title = "Biochemical characterization of the functional roles of residues in the active site of the β-galactosidase from Bacillus circulans ATCC 31382",
abstract = "The β-galactosidase enzyme from Bacillus circulans ATCC 31382 BgaD is widely used in the food industry to produce prebiotic galactooligosaccharides (GOS). Recently, the crystal structure of a C-terminally truncated version of the enzyme (BgaD-D) has been elucidated. The roles of active site amino acid residues in β-galactosidase enzyme reaction and product specificity have remained unknown. Based on a structural alignment of the β-galactosidase enzymes BgaD-D from Bacillus circulans and BgaA from Streptococcus pneumoniae, and the complex of BgaA with LacNAc, we identified 8 active site amino acid residues (Arg185, Asp481, Lys487, Tyr511, Trp570, Trp593, Glu601, and Phe616) in BgaD-D. This study reports an investigation of the functional roles of these residues, using site-directed mutagenesis, and a detailed biochemical characterization and product profile analysis of the mutants obtained. The data shows that these residues are involved in binding and positioning of the substrate, and thus determine the BgaD-D activity and product linkage specificity. This study gives detailed insights into structure-function relationships of the B. circulans BgaD-D enzyme, especially regarding GOS product linkage specificity, allowing the rational mutation of β-galactosidase enzymes to produce specific mixtures of GOS structures.",
keywords = "LACTOSE, ENZYME, HUMAN COLONIC MICROBIOTA, GALACTOOLIGOSACCHARIDE PRODUCTION, FRUCTO-OLIGOSACCHARIDES, KLUYVEROMYCES-LACTIS, DIETARY MODULATION, CRYSTAL-STRUCTURE, PREBIOTICS, SPECIFICITY",
author = "Huifang Yin and Tjaard Pijning and Xiangfeng Meng and Lubbert Dijkhuizen and {van Leeuwen}, {Sander Sebastiaan}",
year = "2017",
doi = "10.1021/acs.biochem.7b00207",
language = "English",
volume = "56",
pages = "3109--3118",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "AMER CHEMICAL SOC",
number = "24",

}

RIS

TY - JOUR

T1 - Biochemical characterization of the functional roles of residues in the active site of the β-galactosidase from Bacillus circulans ATCC 31382

AU - Yin, Huifang

AU - Pijning, Tjaard

AU - Meng, Xiangfeng

AU - Dijkhuizen, Lubbert

AU - van Leeuwen, Sander Sebastiaan

PY - 2017

Y1 - 2017

N2 - The β-galactosidase enzyme from Bacillus circulans ATCC 31382 BgaD is widely used in the food industry to produce prebiotic galactooligosaccharides (GOS). Recently, the crystal structure of a C-terminally truncated version of the enzyme (BgaD-D) has been elucidated. The roles of active site amino acid residues in β-galactosidase enzyme reaction and product specificity have remained unknown. Based on a structural alignment of the β-galactosidase enzymes BgaD-D from Bacillus circulans and BgaA from Streptococcus pneumoniae, and the complex of BgaA with LacNAc, we identified 8 active site amino acid residues (Arg185, Asp481, Lys487, Tyr511, Trp570, Trp593, Glu601, and Phe616) in BgaD-D. This study reports an investigation of the functional roles of these residues, using site-directed mutagenesis, and a detailed biochemical characterization and product profile analysis of the mutants obtained. The data shows that these residues are involved in binding and positioning of the substrate, and thus determine the BgaD-D activity and product linkage specificity. This study gives detailed insights into structure-function relationships of the B. circulans BgaD-D enzyme, especially regarding GOS product linkage specificity, allowing the rational mutation of β-galactosidase enzymes to produce specific mixtures of GOS structures.

AB - The β-galactosidase enzyme from Bacillus circulans ATCC 31382 BgaD is widely used in the food industry to produce prebiotic galactooligosaccharides (GOS). Recently, the crystal structure of a C-terminally truncated version of the enzyme (BgaD-D) has been elucidated. The roles of active site amino acid residues in β-galactosidase enzyme reaction and product specificity have remained unknown. Based on a structural alignment of the β-galactosidase enzymes BgaD-D from Bacillus circulans and BgaA from Streptococcus pneumoniae, and the complex of BgaA with LacNAc, we identified 8 active site amino acid residues (Arg185, Asp481, Lys487, Tyr511, Trp570, Trp593, Glu601, and Phe616) in BgaD-D. This study reports an investigation of the functional roles of these residues, using site-directed mutagenesis, and a detailed biochemical characterization and product profile analysis of the mutants obtained. The data shows that these residues are involved in binding and positioning of the substrate, and thus determine the BgaD-D activity and product linkage specificity. This study gives detailed insights into structure-function relationships of the B. circulans BgaD-D enzyme, especially regarding GOS product linkage specificity, allowing the rational mutation of β-galactosidase enzymes to produce specific mixtures of GOS structures.

KW - LACTOSE

KW - ENZYME

KW - HUMAN COLONIC MICROBIOTA

KW - GALACTOOLIGOSACCHARIDE PRODUCTION

KW - FRUCTO-OLIGOSACCHARIDES

KW - KLUYVEROMYCES-LACTIS

KW - DIETARY MODULATION

KW - CRYSTAL-STRUCTURE

KW - PREBIOTICS

KW - SPECIFICITY

U2 - 10.1021/acs.biochem.7b00207

DO - 10.1021/acs.biochem.7b00207

M3 - Article

C2 - 28538097

VL - 56

SP - 3109

EP - 3118

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 24

ER -

ID: 42504117