Biocatalytic production of 3′-sialyllactose by use of a modified sialidase with superior trans-sialidase activity

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

Biocatalytic production of 3′-sialyllactose by use of a modified sialidase with superior trans-sialidase activity. / Michalak, Malwina; Larsen, Dorte M.; Jers, Carsten; Almeida, João R.M.; Willer, Martin; Li, Haiying; Kirpekar, Finn; Kjærulff, Louise; Gotfredsen, Charlotte H.; Nordvang, Rune T.; Meyer, Anne S.; Mikkelsen, Jørn D.

In: Process Biochemistry, Vol. 49, No. 2, 2014, p. 265-270.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Michalak, M, Larsen, DM, Jers, C, Almeida, JRM, Willer, M, Li, H, Kirpekar, F, Kjærulff, L, Gotfredsen, CH, Nordvang, RT, Meyer, AS & Mikkelsen, JD 2014, 'Biocatalytic production of 3′-sialyllactose by use of a modified sialidase with superior trans-sialidase activity', Process Biochemistry, vol. 49, no. 2, pp. 265-270. https://doi.org/10.1016/j.procbio.2013.10.023

APA

Michalak, M., Larsen, D. M., Jers, C., Almeida, J. R. M., Willer, M., Li, H., Kirpekar, F., Kjærulff, L., Gotfredsen, C. H., Nordvang, R. T., Meyer, A. S., & Mikkelsen, J. D. (2014). Biocatalytic production of 3′-sialyllactose by use of a modified sialidase with superior trans-sialidase activity. Process Biochemistry, 49(2), 265-270. https://doi.org/10.1016/j.procbio.2013.10.023

Vancouver

Michalak M, Larsen DM, Jers C, Almeida JRM, Willer M, Li H et al. Biocatalytic production of 3′-sialyllactose by use of a modified sialidase with superior trans-sialidase activity. Process Biochemistry. 2014;49(2):265-270. https://doi.org/10.1016/j.procbio.2013.10.023

Author

Michalak, Malwina ; Larsen, Dorte M. ; Jers, Carsten ; Almeida, João R.M. ; Willer, Martin ; Li, Haiying ; Kirpekar, Finn ; Kjærulff, Louise ; Gotfredsen, Charlotte H. ; Nordvang, Rune T. ; Meyer, Anne S. ; Mikkelsen, Jørn D. / Biocatalytic production of 3′-sialyllactose by use of a modified sialidase with superior trans-sialidase activity. In: Process Biochemistry. 2014 ; Vol. 49, No. 2. pp. 265-270.

Bibtex

@article{0d5959b61b93450fb785cbd55382f90f,
title = "Biocatalytic production of 3′-sialyllactose by use of a modified sialidase with superior trans-sialidase activity",
abstract = "Casein glycomacropeptide (cGMP) and lactose, which are purified (or semi-purified) components obtained from side streams from dairy industry operations, were used as substrates for enzyme catalyzed production of 3′-sialyllactose, a model case compound for human milk oligosaccharides (HMOs). The enzyme employed was a mutated sialidase, Tr6, derived from Trypanosoma rangeli, and expressed in Pichia pastoris after codon-optimization. The Tr6 contained 6 point mutations and exhibited trans-sialidase activity. The Tr6 trans-sialidase reaction conditions were tuned for maximizing Tr6 catalyzed 3′-sialyllactose production by optimizing pH, temperature, acceptor, and donor concentrations using response surface designs. At the optimum reaction conditions, the Tr6 catalyzed the transfer of sialic acid from cGMP to lactose at high efficiency without substantial hydrolysis of the 3′-sialyllactose product. The robustness of the Tr6 catalyzed reaction was verified at 5 L-scale providing a yield of 3.6 g 3′-sialyllactose at an estimated molar trans-sialylation yield of 50% on the 3′-sialyl in cGMP. Lacto-N-tetraose and lacto-N-fucopentaoses also functioned as acceptor molecules demonstrating the versatility of the Tr6 trans-sialidase for catalyzing sialyl-transfer for generating different HMOs. The data signify the applicability of enzymatic trans-sialylation on dairy side-stream components for production of human milk oligosaccharides.",
keywords = "3′-Sialyllactose, Human milk oligosaccharides, trans-Sialylation, Trypanosoma rangeli",
author = "Malwina Michalak and Larsen, {Dorte M.} and Carsten Jers and Almeida, {Jo{\~a}o R.M.} and Martin Willer and Haiying Li and Finn Kirpekar and Louise Kj{\ae}rulff and Gotfredsen, {Charlotte H.} and Nordvang, {Rune T.} and Meyer, {Anne S.} and Mikkelsen, {J{\o}rn D.}",
year = "2014",
doi = "10.1016/j.procbio.2013.10.023",
language = "English",
volume = "49",
pages = "265--270",
journal = "Process Biochemistry",
issn = "0032-9592",
publisher = "Elsevier",
number = "2",

}

RIS

TY - JOUR

T1 - Biocatalytic production of 3′-sialyllactose by use of a modified sialidase with superior trans-sialidase activity

AU - Michalak, Malwina

AU - Larsen, Dorte M.

AU - Jers, Carsten

AU - Almeida, João R.M.

AU - Willer, Martin

AU - Li, Haiying

AU - Kirpekar, Finn

AU - Kjærulff, Louise

AU - Gotfredsen, Charlotte H.

AU - Nordvang, Rune T.

AU - Meyer, Anne S.

AU - Mikkelsen, Jørn D.

PY - 2014

Y1 - 2014

N2 - Casein glycomacropeptide (cGMP) and lactose, which are purified (or semi-purified) components obtained from side streams from dairy industry operations, were used as substrates for enzyme catalyzed production of 3′-sialyllactose, a model case compound for human milk oligosaccharides (HMOs). The enzyme employed was a mutated sialidase, Tr6, derived from Trypanosoma rangeli, and expressed in Pichia pastoris after codon-optimization. The Tr6 contained 6 point mutations and exhibited trans-sialidase activity. The Tr6 trans-sialidase reaction conditions were tuned for maximizing Tr6 catalyzed 3′-sialyllactose production by optimizing pH, temperature, acceptor, and donor concentrations using response surface designs. At the optimum reaction conditions, the Tr6 catalyzed the transfer of sialic acid from cGMP to lactose at high efficiency without substantial hydrolysis of the 3′-sialyllactose product. The robustness of the Tr6 catalyzed reaction was verified at 5 L-scale providing a yield of 3.6 g 3′-sialyllactose at an estimated molar trans-sialylation yield of 50% on the 3′-sialyl in cGMP. Lacto-N-tetraose and lacto-N-fucopentaoses also functioned as acceptor molecules demonstrating the versatility of the Tr6 trans-sialidase for catalyzing sialyl-transfer for generating different HMOs. The data signify the applicability of enzymatic trans-sialylation on dairy side-stream components for production of human milk oligosaccharides.

AB - Casein glycomacropeptide (cGMP) and lactose, which are purified (or semi-purified) components obtained from side streams from dairy industry operations, were used as substrates for enzyme catalyzed production of 3′-sialyllactose, a model case compound for human milk oligosaccharides (HMOs). The enzyme employed was a mutated sialidase, Tr6, derived from Trypanosoma rangeli, and expressed in Pichia pastoris after codon-optimization. The Tr6 contained 6 point mutations and exhibited trans-sialidase activity. The Tr6 trans-sialidase reaction conditions were tuned for maximizing Tr6 catalyzed 3′-sialyllactose production by optimizing pH, temperature, acceptor, and donor concentrations using response surface designs. At the optimum reaction conditions, the Tr6 catalyzed the transfer of sialic acid from cGMP to lactose at high efficiency without substantial hydrolysis of the 3′-sialyllactose product. The robustness of the Tr6 catalyzed reaction was verified at 5 L-scale providing a yield of 3.6 g 3′-sialyllactose at an estimated molar trans-sialylation yield of 50% on the 3′-sialyl in cGMP. Lacto-N-tetraose and lacto-N-fucopentaoses also functioned as acceptor molecules demonstrating the versatility of the Tr6 trans-sialidase for catalyzing sialyl-transfer for generating different HMOs. The data signify the applicability of enzymatic trans-sialylation on dairy side-stream components for production of human milk oligosaccharides.

KW - 3′-Sialyllactose

KW - Human milk oligosaccharides

KW - trans-Sialylation

KW - Trypanosoma rangeli

U2 - 10.1016/j.procbio.2013.10.023

DO - 10.1016/j.procbio.2013.10.023

M3 - Journal article

AN - SCOPUS:84895069393

VL - 49

SP - 265

EP - 270

JO - Process Biochemistry

JF - Process Biochemistry

SN - 0032-9592

IS - 2

ER -

ID: 192043729