The human digestive system consists of a complex series of organs and glands regulating the processing of food. The process starts in the mouth, where the food breaks down by chewing, and the digestive enzymes in the saliva. Then in the stomach the gastric phase of digestion takes place. At the final stage the food is mixed with enzymes produced by the pancreas, after which the content passes the gastrointestinal tract. The undigested food particles end up in the large intestine, where they are fermented by the gut microbiome, consisting of approximately thousand bacterial species. CarbExplore is an expert in manufacturing, purification and analysis of prebiotic food compounds, such as Fructo-oligosaccharides (FOS), Galacto-oligosaccharides (GOS), Isomalto-oligosaccharides (IMO), Isomalto-/maltopolysaccharides (IMMP), milk oligosaccharides, and N-glycans of milk proteins. These potential prebiotic food components support the growth of the microorganisms that provide a health benefit.

CarbExplore has a library with a broad range of Carbohydrate-Active Enzymes, e.g. glucansucrase enzymes (dextransucrases, mutansucrases, reuteransucrases, alternansucrases), alpha-glucanotransferases, beta-galactosidases and fructansucrase enzymes (inulosucrases, levansucrases). These enzymes are able to convert substrate molecules into the products of interest. We are able to enzymatically modify the carbohydrate structures and functions of the targeted substrates, introducing changes in the degree of polymerization (DP), branching degree, and glycosidic linkages present (α1→2), (α1→3), (α1→4) and (α1→6) (in linear or alternating chains). Our scientists use several techniques (HPLC, HPAEC-PAD, NMR, methylation analysis, MALDI-TOF, HP-SEC) to structurally analyze the products, and to isolate the products of interest.

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Examples:

✔ α-Glucan polymers or oligosaccharides with different DP values and linkage types, synthesized from sucrose using glucansucrase wild type and mutant enzymes

✔ Fructan polymers (inulin, levan) or oligosaccharides with different DP values and linkage types, synthesized from sucrose using fructansucrase wild type and mutant enzymes

✔ Mixtures of GOS with different DP values and/or linkage types, synthesized from lactose using β-galactosidase wild type and mutant enzymes

✔ IMO and/or IMMP synthesized from starch (amylose and/or amylopectin) using 4,6-α-glucanotransferase wild type and mutant enzymes

✔ (α1→3)-branched IMO and/or IMMP, using branching sucrase enzymes and sucrose

✔Synthesis, purification and characterization of milk oligosaccharides

✔Large-scale quantitative isolation of pure N-linked glycans of milk proteins

Workflow:


You deliver:

Detailed project information about the product of interest.


We deliver:

A project plan with detailed descriptions.

Our Relevant Publications:

Publications 2020

Gangoiti, J. et al., 2020. Synthesis of novel α-glucans with potential health benefits through controlled glucose release in the human gastrointestinal tract. Critical Reviews in Food Science and Nutrition, 2 1, 60(1), pp. 123-146. DOI 10.1080/10408398.2018.1516621

te Poele, E. et al., 2020. Development of slowly digestible starch derived alpha-glucans with 4,6-α-glucanotransferase and branching sucrase enzymes. Journal of Agricultural and Food Chemistry, 21 5.p. acs.jafc.0c01465. DOI 10.1021/acs.jafc.0c01465

Publications 2019

Pham, H. T., Ten Kate, G. A., Dijkhuizen, L. & Van Leeuwen, S. S., 2019. Synthesis and Characterization of Sialylated Lactose- and Lactulose-Derived Oligosaccharides by Trypanosoma cruzi Trans-sialidase. Journal of Agricultural and Food Chemistry, 27 3, 67(12), pp. 3469-3479. DOI 10.1021/acs.jafc.8b06974

Valk-Weeber, R. L., Dijkhuizen, L. & van Leeuwen, S. S., 2019. Large-scale quantitative isolation of pure protein N-linked glycans. Carbohydrate Research, 1 6, Volume 479, pp. 13-22. DOI 10.1016/j.carres.2019.04.011

Publications 2018

Gangoiti, J., Pijning, T. & Dijkhuizen, L., 2018. Biotechnological potential of novel glycoside hydrolase family 70 enzymes synthesizing α-glucans from starch and sucrose. Biotechnology Advances, 36(1), pp. 196-207. DOI 10.1016/j.biotechadv.2017.11.001

Pham, H., Pijning, T., Dijkhuizen, L. & Van Leeuwen, S. S., 2018. Mutational analysis of the role of the glucansucrase Gtf180-ΔN active site residues in product and linkage specificity with lactose as acceptor substrate. Journal of Agricultural and Food Chemistry, 28 11, 66(47), pp. 12544-12554. DOI 10.1021/acs.jafc.8b04486

Pham, H. T., Dijkhuizen, L. & van Leeuwen, S. S., 2018. Structural characterization of glucosylated GOS derivatives synthesized by the Lactobacillus reuteri GtfA and Gtf180 glucansucrase enzymes. Carbohydrate Research, 1 12, Volume 470, pp. 57-63. DOI 10.1016/j.carres.2018.10.003

van Leeuwen, S. S. et al., 2018. Regional variations in human milk oligosaccharides in Vietnam suggest FucTx activity besides FucT2 and FucT3. Scientific Reports, 1 12.8(16790). DOI 10.1038/s41598-018-34882-x

Yin, H., Dijkhuizen, L. & van Leeuwen, S. S., 2018. Synthesis of galacto-oligosaccharides derived from lactulose by wild-type and mutant β-galactosidase enzymes from Bacillus circulans ATCC 31382. Carbohydrate Research, 30 7, Volume 465, pp. 58-65. DOI 10.1016/j.carres.2018.06.009

Figueroa-Lozano, S. et al., 2018. Dietary N-Glycans from Bovine Lactoferrin and TLR Modulation. Molecular Nutrition and Food Research, 1 1, 62(2), p. 1700389. DOI 10.1002/mnfr.201700389

Publications 2017

Pham, H. T., Dijkhuizen, L. & van Leeuwen, S. S., 2017. Structural characterization of glucosylated lactose derivatives synthesized by the Lactobacillus reuteri GtfA and Gtf180 glucansucrase enzymes. Carbohydrate Research, Volume 449, pp. 59-64. DOI 10.1016/j.carres.2017.07.002

Yin, H. et al., 2017. Engineering of the Bacillus circulans β-galactosidase product specificity. Biochemistry, 7 2, 56(5), pp. 704-711. DOI 10.1021/acs.biochem.7b00032

Publications 2016

Meng, X. et al., 2016. Structure–function relationships of family GH70 glucansucrase and 4,6-α-glucanotransferase enzymes, and their evolutionary relationships with family GH13 enzymes. s.l.:Birkhauser Verlag AG. DOI 10.1007/s00018-016-2245-7

Publications 2015

Wilbrink, M. H. et al., 2015. Enzymatic decoration of prebiotic galacto-oligosaccharides (Vivinal GOS) with sialic acid using Trypanosoma cruzi trans -sialidase and two bovine sialoglycoconjugates as donor substrates. Journal of Agricultural and Food Chemistry, 1 7, 63(25), pp. 5976-5984. DOI 10.1021/acs.jafc.5b01505

Leemhuis, H. et al., 2014. Isomalto/malto-polysaccharide, a novel soluble dietary fiber made via enzymatic conversion of starch. Journal of Agricultural and Food Chemistry, 10 12, 62(49), pp. 12034-12044. DOI 10.1021/jf503970a

Publications 2014

Wilbrink, M. H. et al., 2014. Galactosyl-lactose sialylation using Trypanosoma cruzi trans-sialidase as the biocatalyst and bovine κ-casein-derived glycomacropeptide asthe donor substrate. Applied and Environmental Microbiology, 80(19), pp. 5984-5991. DOI 10.1128/AEM.01465-14

Publications 2013

Leemhuis, H. et al., 2013. Glucansucrases: Three-dimensional structures, reactions, mechanism, α-glucan analysis and their implications in biotechnology and food applications. Journal of Biotechnology, 1, 163(2), pp. 250-272. DOI 10.1016/j.jbiotec.2012.06.037

Publications 2012

Anwar, M. A. et al., 2012. The role of conserved inulosucrase residues in the reaction and product specificity of Lactobacillus reuteri inulosucrase. FEBS Journal, 10, 279(19), pp. 3612-3621. DOI 10.1111/j.1742-4658.2012.08721.x

Desmet, T. et al., 2012. Enzymatic glycosylation of small molecules: Challenging substrates require tailored catalysts. Chemistry – A European Journal, 27 8, 18(35), pp. 10786-10801. DOI 10.1002/chem.201103069

Publications 2011

Kralj, S. et al., 2011. 4,6-α-glucanotransferase, a novel enzyme that structurally and functionally provides an evolutionary link between glycoside hydrolase enzyme families 13 and 70. Applied and Environmental Microbiology, 11, 77(22), pp. 8154-8163. DOI 10.1128/AEM.05735-11