Publication

Biochemical characterization of Lactobacillus reuteri Glycoside Hydrolase family 70 GTFB type of 4,6-α-Glucanotransferase enzymes that synthesize soluble dietary starch fibers

Bai, Y., van der Kaaij, R. M., Leemhuis, H., Pijning, T., van Leeuwen, S. S., Jin, Z. & Dijkhuizen, L., 7-Aug-2015, In : Applied and environmental microbiology. 81, 20, p. 7223-7232 10 p.

Research output: Contribution to journalArticleAcademicpeer-review

APA

Bai, Y., van der Kaaij, R. M., Leemhuis, H., Pijning, T., van Leeuwen, S. S., Jin, Z., & Dijkhuizen, L. (2015). Biochemical characterization of Lactobacillus reuteri Glycoside Hydrolase family 70 GTFB type of 4,6-α-Glucanotransferase enzymes that synthesize soluble dietary starch fibers. Applied and environmental microbiology, 81(20), 7223-7232. https://doi.org/10.1128/AEM.01860-15

Author

Bai, Yuxiang ; van der Kaaij, Rachel Maria ; Leemhuis, Hans ; Pijning, Tjaard ; van Leeuwen, Sander Sebastiaan ; Jin, Zhengyu ; Dijkhuizen, Lubbert. / Biochemical characterization of Lactobacillus reuteri Glycoside Hydrolase family 70 GTFB type of 4,6-α-Glucanotransferase enzymes that synthesize soluble dietary starch fibers. In: Applied and environmental microbiology. 2015 ; Vol. 81, No. 20. pp. 7223-7232.

Harvard

Bai, Y, van der Kaaij, RM, Leemhuis, H, Pijning, T, van Leeuwen, SS, Jin, Z & Dijkhuizen, L 2015, 'Biochemical characterization of Lactobacillus reuteri Glycoside Hydrolase family 70 GTFB type of 4,6-α-Glucanotransferase enzymes that synthesize soluble dietary starch fibers', Applied and environmental microbiology, vol. 81, no. 20, pp. 7223-7232. https://doi.org/10.1128/AEM.01860-15

Standard

Biochemical characterization of Lactobacillus reuteri Glycoside Hydrolase family 70 GTFB type of 4,6-α-Glucanotransferase enzymes that synthesize soluble dietary starch fibers. / Bai, Yuxiang; van der Kaaij, Rachel Maria; Leemhuis, Hans; Pijning, Tjaard; van Leeuwen, Sander Sebastiaan; Jin, Zhengyu; Dijkhuizen, Lubbert.

In: Applied and environmental microbiology, Vol. 81, No. 20, 07.08.2015, p. 7223-7232.

Research output: Contribution to journalArticleAcademicpeer-review

Vancouver

Bai Y, van der Kaaij RM, Leemhuis H, Pijning T, van Leeuwen SS, Jin Z et al. Biochemical characterization of Lactobacillus reuteri Glycoside Hydrolase family 70 GTFB type of 4,6-α-Glucanotransferase enzymes that synthesize soluble dietary starch fibers. Applied and environmental microbiology. 2015 Aug 7;81(20):7223-7232. https://doi.org/10.1128/AEM.01860-15


BibTeX

@article{d956110915cd4115a6dac8ab45544aee,
title = "Biochemical characterization of Lactobacillus reuteri Glycoside Hydrolase family 70 GTFB type of 4,6-α-Glucanotransferase enzymes that synthesize soluble dietary starch fibers",
abstract = "4,6-α-Glucanotransferase (4,6-α-GTase) enzymes, such as GTFB and GTFW of Lactobacillus reuteri strains, constitute a new reaction specificity in Glycoside Hydrolase Family 70 (GH70) and are novel enzymes that convert starch or starch hydrolysates into isomalto/malto-polysaccharides (IMMPs). These IMMPs still have linear chains with some α1→4 linkages but mostly (relatively long) linear chains with α1→6 linkages, and are soluble dietary starch fibers. 4,6-α-GTase enzymes and their products have significant potential for industrial applications. Here we report that an N-terminal truncation (1-733 amino acids) strongly enhances the soluble expression level of fully active GTFB-ΔN (approx. 75 fold compared to full length wild type GTFB) in Escherichi coli. In addition, quantitative assays based on amylose V as substrate are described, allowing accurate determination of both hydrolysis (minor) activity (glucose release, reducing power) and total activity (iodine staining), and calculation of the transferase (major) activity of these 4,6-α-GTase enzymes. The data shows that GTFB-ΔN is clearly less hydrolytic than GTFW, which is also supported by NMR analysis of their final products. Using these assays, the biochemical properties of GTFB-ΔN were characterized in detail, including determination of kinetic parameters and acceptor substrate specificity. The GTFB-ΔN enzyme displayed high conversion yields at relatively high substrate concentrations, a promising feature for industrial application.",
author = "Yuxiang Bai and {van der Kaaij}, {Rachel Maria} and Hans Leemhuis and Tjaard Pijning and {van Leeuwen}, {Sander Sebastiaan} and Zhengyu Jin and Lubbert Dijkhuizen",
note = "Copyright {\textcopyright} 2015, American Society for Microbiology. All Rights Reserved.",
year = "2015",
month = aug,
day = "7",
doi = "10.1128/AEM.01860-15",
language = "English",
volume = "81",
pages = "7223--7232",
journal = "Applied Environmental Microbiology",
issn = "0099-2240",
publisher = "AMER SOC MICROBIOLOGY",
number = "20",

}

RIS

TY - JOUR

T1 - Biochemical characterization of Lactobacillus reuteri Glycoside Hydrolase family 70 GTFB type of 4,6-α-Glucanotransferase enzymes that synthesize soluble dietary starch fibers

AU - Bai, Yuxiang

AU - van der Kaaij, Rachel Maria

AU - Leemhuis, Hans

AU - Pijning, Tjaard

AU - van Leeuwen, Sander Sebastiaan

AU - Jin, Zhengyu

AU - Dijkhuizen, Lubbert

N1 - Copyright © 2015, American Society for Microbiology. All Rights Reserved.

PY - 2015/8/7

Y1 - 2015/8/7

N2 - 4,6-α-Glucanotransferase (4,6-α-GTase) enzymes, such as GTFB and GTFW of Lactobacillus reuteri strains, constitute a new reaction specificity in Glycoside Hydrolase Family 70 (GH70) and are novel enzymes that convert starch or starch hydrolysates into isomalto/malto-polysaccharides (IMMPs). These IMMPs still have linear chains with some α1→4 linkages but mostly (relatively long) linear chains with α1→6 linkages, and are soluble dietary starch fibers. 4,6-α-GTase enzymes and their products have significant potential for industrial applications. Here we report that an N-terminal truncation (1-733 amino acids) strongly enhances the soluble expression level of fully active GTFB-ΔN (approx. 75 fold compared to full length wild type GTFB) in Escherichi coli. In addition, quantitative assays based on amylose V as substrate are described, allowing accurate determination of both hydrolysis (minor) activity (glucose release, reducing power) and total activity (iodine staining), and calculation of the transferase (major) activity of these 4,6-α-GTase enzymes. The data shows that GTFB-ΔN is clearly less hydrolytic than GTFW, which is also supported by NMR analysis of their final products. Using these assays, the biochemical properties of GTFB-ΔN were characterized in detail, including determination of kinetic parameters and acceptor substrate specificity. The GTFB-ΔN enzyme displayed high conversion yields at relatively high substrate concentrations, a promising feature for industrial application.

AB - 4,6-α-Glucanotransferase (4,6-α-GTase) enzymes, such as GTFB and GTFW of Lactobacillus reuteri strains, constitute a new reaction specificity in Glycoside Hydrolase Family 70 (GH70) and are novel enzymes that convert starch or starch hydrolysates into isomalto/malto-polysaccharides (IMMPs). These IMMPs still have linear chains with some α1→4 linkages but mostly (relatively long) linear chains with α1→6 linkages, and are soluble dietary starch fibers. 4,6-α-GTase enzymes and their products have significant potential for industrial applications. Here we report that an N-terminal truncation (1-733 amino acids) strongly enhances the soluble expression level of fully active GTFB-ΔN (approx. 75 fold compared to full length wild type GTFB) in Escherichi coli. In addition, quantitative assays based on amylose V as substrate are described, allowing accurate determination of both hydrolysis (minor) activity (glucose release, reducing power) and total activity (iodine staining), and calculation of the transferase (major) activity of these 4,6-α-GTase enzymes. The data shows that GTFB-ΔN is clearly less hydrolytic than GTFW, which is also supported by NMR analysis of their final products. Using these assays, the biochemical properties of GTFB-ΔN were characterized in detail, including determination of kinetic parameters and acceptor substrate specificity. The GTFB-ΔN enzyme displayed high conversion yields at relatively high substrate concentrations, a promising feature for industrial application.

U2 - 10.1128/AEM.01860-15

DO - 10.1128/AEM.01860-15

M3 - Article

C2 - 26253678

VL - 81

SP - 7223

EP - 7232

JO - Applied Environmental Microbiology

JF - Applied Environmental Microbiology

SN - 0099-2240

IS - 20

ER -

ID: 22890988