Publication

Mutational Analysis of the Role of the Glucansucrase Gtf180-Delta N Active Site Residues in Product and Linkage Specificity with Lactose as Acceptor Substrate

Pham, H., Pijning, T., Dijkhuizen, L. & van Leeuwen, S. S., 28-Nov-2018, In : Journal of Agricultural and Food Chemistry. 66, 47, p. 12544-12554 11 p., acs.jafc.8b04486.

Research output: Contribution to journalArticleAcademicpeer-review

APA

Pham, H., Pijning, T., Dijkhuizen, L., & van Leeuwen, S. S. (2018). Mutational Analysis of the Role of the Glucansucrase Gtf180-Delta N Active Site Residues in Product and Linkage Specificity with Lactose as Acceptor Substrate. Journal of Agricultural and Food Chemistry, 66(47), 12544-12554. [acs.jafc.8b04486]. https://doi.org/10.1021/acs.jafc.8b04486

Author

Pham, Hien ; Pijning, Tjaard ; Dijkhuizen, Lubbert ; van Leeuwen, Sander Sebastiaan. / Mutational Analysis of the Role of the Glucansucrase Gtf180-Delta N Active Site Residues in Product and Linkage Specificity with Lactose as Acceptor Substrate. In: Journal of Agricultural and Food Chemistry. 2018 ; Vol. 66, No. 47. pp. 12544-12554.

Harvard

Pham, H, Pijning, T, Dijkhuizen, L & van Leeuwen, SS 2018, 'Mutational Analysis of the Role of the Glucansucrase Gtf180-Delta N Active Site Residues in Product and Linkage Specificity with Lactose as Acceptor Substrate', Journal of Agricultural and Food Chemistry, vol. 66, no. 47, acs.jafc.8b04486, pp. 12544-12554. https://doi.org/10.1021/acs.jafc.8b04486

Standard

Mutational Analysis of the Role of the Glucansucrase Gtf180-Delta N Active Site Residues in Product and Linkage Specificity with Lactose as Acceptor Substrate. / Pham, Hien; Pijning, Tjaard; Dijkhuizen, Lubbert; van Leeuwen, Sander Sebastiaan.

In: Journal of Agricultural and Food Chemistry, Vol. 66, No. 47, acs.jafc.8b04486, 28.11.2018, p. 12544-12554.

Research output: Contribution to journalArticleAcademicpeer-review

Vancouver

Pham H, Pijning T, Dijkhuizen L, van Leeuwen SS. Mutational Analysis of the Role of the Glucansucrase Gtf180-Delta N Active Site Residues in Product and Linkage Specificity with Lactose as Acceptor Substrate. Journal of Agricultural and Food Chemistry. 2018 Nov 28;66(47):12544-12554. acs.jafc.8b04486. https://doi.org/10.1021/acs.jafc.8b04486


BibTeX

@article{0d4f6b8c226d4b709426aa549d160aec,
title = "Mutational Analysis of the Role of the Glucansucrase Gtf180-Delta N Active Site Residues in Product and Linkage Specificity with Lactose as Acceptor Substrate",
abstract = "Glucansucrase Gtf180-Delta N from Lactobacillus reuteri uses lactose as acceptor substrate to synthesize five glucosylated lactose molecules (F1-F5) with a degree of polymerization (DP) of 3-4 (GL34) and with (alpha 1 -> 2)/(alpha 1 -> 3)/(alpha 1 -> 4) glycosidic linkages. Q1140/W1065/N1029 mutations significantly changed the GL34 product ratios. Q1140 mutations clearly decreased F3 3'-glc-lac with an (alpha 1 -> 3) linkage and increased F4 4',2-glc-lac with (alpha 1 -> 4)/(alpha 1 -> 2) linkages. Formation of F2 2-glc-lac with an (alpha 1 -> 2) linkage and F4 was negatively affected in most W1065 and N1029 mutants, respectively. Mutant N1029G synthesized four new products with additional (alpha 1 -> 3)-linked glucosyl moieties (2xDP4 and 2xDP5). Sucrose/lactose strongly reduced Gtf180-Delta N hydrolytic activity and increased transferase activity of Gtf180-Delta N and mutant N1029G, in comparison to activity with sucrose alone. N1029/W1065/Q1140 thus are key determinants of Gtf180-Delta N linkage and product specificity in the acceptor reaction with lactose. Mutagenesis of key residues in Gtf180-Delta N may allow synthesis of tailor-made mixtures of novel lactose-derived oligosaccharides with potential applications as prebiotic compounds in food/feed and in pharmacy/medicine.",
author = "Hien Pham and Tjaard Pijning and Lubbert Dijkhuizen and {van Leeuwen}, {Sander Sebastiaan}",
year = "2018",
month = nov,
day = "28",
doi = "10.1021/acs.jafc.8b04486",
language = "English",
volume = "66",
pages = "12544--12554",
journal = "Journal of Agricultural and Food Chemistry",
issn = "0021-8561",
publisher = "AMER CHEMICAL SOC",
number = "47",

}

RIS

TY - JOUR

T1 - Mutational Analysis of the Role of the Glucansucrase Gtf180-Delta N Active Site Residues in Product and Linkage Specificity with Lactose as Acceptor Substrate

AU - Pham, Hien

AU - Pijning, Tjaard

AU - Dijkhuizen, Lubbert

AU - van Leeuwen, Sander Sebastiaan

PY - 2018/11/28

Y1 - 2018/11/28

N2 - Glucansucrase Gtf180-Delta N from Lactobacillus reuteri uses lactose as acceptor substrate to synthesize five glucosylated lactose molecules (F1-F5) with a degree of polymerization (DP) of 3-4 (GL34) and with (alpha 1 -> 2)/(alpha 1 -> 3)/(alpha 1 -> 4) glycosidic linkages. Q1140/W1065/N1029 mutations significantly changed the GL34 product ratios. Q1140 mutations clearly decreased F3 3'-glc-lac with an (alpha 1 -> 3) linkage and increased F4 4',2-glc-lac with (alpha 1 -> 4)/(alpha 1 -> 2) linkages. Formation of F2 2-glc-lac with an (alpha 1 -> 2) linkage and F4 was negatively affected in most W1065 and N1029 mutants, respectively. Mutant N1029G synthesized four new products with additional (alpha 1 -> 3)-linked glucosyl moieties (2xDP4 and 2xDP5). Sucrose/lactose strongly reduced Gtf180-Delta N hydrolytic activity and increased transferase activity of Gtf180-Delta N and mutant N1029G, in comparison to activity with sucrose alone. N1029/W1065/Q1140 thus are key determinants of Gtf180-Delta N linkage and product specificity in the acceptor reaction with lactose. Mutagenesis of key residues in Gtf180-Delta N may allow synthesis of tailor-made mixtures of novel lactose-derived oligosaccharides with potential applications as prebiotic compounds in food/feed and in pharmacy/medicine.

AB - Glucansucrase Gtf180-Delta N from Lactobacillus reuteri uses lactose as acceptor substrate to synthesize five glucosylated lactose molecules (F1-F5) with a degree of polymerization (DP) of 3-4 (GL34) and with (alpha 1 -> 2)/(alpha 1 -> 3)/(alpha 1 -> 4) glycosidic linkages. Q1140/W1065/N1029 mutations significantly changed the GL34 product ratios. Q1140 mutations clearly decreased F3 3'-glc-lac with an (alpha 1 -> 3) linkage and increased F4 4',2-glc-lac with (alpha 1 -> 4)/(alpha 1 -> 2) linkages. Formation of F2 2-glc-lac with an (alpha 1 -> 2) linkage and F4 was negatively affected in most W1065 and N1029 mutants, respectively. Mutant N1029G synthesized four new products with additional (alpha 1 -> 3)-linked glucosyl moieties (2xDP4 and 2xDP5). Sucrose/lactose strongly reduced Gtf180-Delta N hydrolytic activity and increased transferase activity of Gtf180-Delta N and mutant N1029G, in comparison to activity with sucrose alone. N1029/W1065/Q1140 thus are key determinants of Gtf180-Delta N linkage and product specificity in the acceptor reaction with lactose. Mutagenesis of key residues in Gtf180-Delta N may allow synthesis of tailor-made mixtures of novel lactose-derived oligosaccharides with potential applications as prebiotic compounds in food/feed and in pharmacy/medicine.

U2 - 10.1021/acs.jafc.8b04486

DO - 10.1021/acs.jafc.8b04486

M3 - Article

C2 - 30396274

VL - 66

SP - 12544

EP - 12554

JO - Journal of Agricultural and Food Chemistry

JF - Journal of Agricultural and Food Chemistry

SN - 0021-8561

IS - 47

M1 - acs.jafc.8b04486

ER -

ID: 67275603