Abstract
The enzymes involved in the biosynthesis of carbohydrates and the attachment of sugar units to biological acceptor molecules catalyse an array of chemical transformations and coupling reactions. In prokaryotes, both common sugar precursors and their enzymatically modified derivatives often become substituents of biologically active natural products through the action of glycosyltransferases. Recently, researchers have begun to harness the power of these biological catalysts to alter the sugar structures and glycosylation patterns of natural products both in vivo and in vitro. Biochemical and structural studies of sugar biosynthetic enzymes and glycosyltransferases, coupled with advances in bioengineering methodology, have ushered in a new era of drug development.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
References
Weymouth-Wilson, A. C. The role of carbohydrates in biologically active natural products. Nat. Prod. Rep. 14, 99–110 (1997).
Thorson, J. S., Hosted, T. J., Jiang, J., Biggins, J. B. & Ahlert, J. Nature's carbohydrate chemists: the enzymatic glycosylation of bioactive bacterial metabolites. Curr. Org. Chem. 5, 139–167 (2001).
Barton, W. A. et al. Structure, mechanism and engineering of a nucleotidylyltransferase as a first step toward glycorandomization. Nature Struct. Biol. 8, 545–551 (2001).
Varki, A. et al. Essentials of Glycobiology (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1999).
Johnson, D. A. & Liu, H.-w. in Comprehensive Natural Product Chemistry (eds Barton, D. H. R., Meth-Cohn, O. & Nakanishi, K.) 311–365 (Elsevier, Amsterdam, 1999).
He, X., Agnihotri, G. & Liu, H.-w. Novel enzymatic mechanisms in carbohydrate metabolism. Chem. Rev. 100, 4615–4661 (2000).
He, X. M. & Liu, H.-w. Formation of unusual sugars: mechanistic studies and biosynthetic applications. Annu. Rev. Biochem. 71, 701–754 (2002).
Hallis, T. M. & Liu, H.-w. Learning Nature's strategies for making deoxy sugars: pathways, mechanisms, and combinatorial applications. Acc. Chem. Res. 32, 579–588 (1999).
He, X. & Liu, H.-w. Mechanisms of enzymatic C–O bond cleavages in deoxyhexose biosynthesis. Curr. Opin. Chem. Biol. 6, 590–597 (2002).
Szu, P.-h., He, X., Zhao, L. & Liu, H.-w. Biosynthesis of TDP-D-desosamine: identification of a strategy for C4 deoxygenation. Angew. Chem. Int. Ed. 44, 6742–6746 (2005).
Oppermann, U. et al. Short-chain dehydrogenases/reductases (SDR): the 2002 update. Chem. Biol. Interact. 143–144, 247–253 (2003).
Frey, P. A. The Leloir pathway: a mechanistic imperative for three enzymes to change the stereochemical configuration of a single carbon in galactose. FASEB J. 10, 461–470 (1996).
Thoden, J. B., Frey, P. A. & Holden, H. M. Molecular structure of the NADH/UDP-glucose abortive complex of UDP-galactose-4-epimerase from Escherichia coli: implications for the catalytic mechanism. Biochemistry 35, 5137–5144 (1996).
Hallis, T. M., Zhao, Z. & Liu, H.-w. New insights into the mechanism of CDP-D-tyvelose 2-epimerase: an enzyme catalyzing epimerization at an unactivated stereocenter. J. Am. Chem. Soc. 122, 10493–10503 (2000).
Morrison, J. P., Read, J. A., Coleman, W. G. & Tanner, M. E. Dismutase activity of ADP-L-glycero-D-manno-heptose 6-epimerase: evidence for a direct oxidation/reduction mechanism. Biochemistry 44, 5907–5915 (2005).
Major, L. L., Wolucka, B. A. & Naismith, J. H. Structure and function of GDP-mannose-3′,5′-epimerase: an enzyme which performs three chemical reactions at the same active site. J. Am. Chem. Soc. 127, 18309–18320 (2005).
Mulichak, A. M., Theisen, M. J., Essigmann, B., Benning, C. & Garavito, R. M. Crystal structure of SQD1, an enzyme involved in the biosynthesis of the plant sulfolipid headgroup donor UDP-sulfoquinovose. Proc. Natl Acad. Sci. USA 96, 13097–13102 (1999).
Sanda, S., Leustek, T., Theisen, M. J., Garavito, R. M. & Benning, C. Recombinant Arabidopsis SQD1 converts UDP-glucose and sulfite to the sulfolipid head group precursor UDP-sulfoquinovose in vitro. J. Biol. Chem. 276, 3941–3946 (2001).
Schutzbach, J. S. & Feingold, D. S. Biosynthesis of uridine diphosphate D-xylose IV. Mechanism of action of uridine diphosphoglucuronate carboxy-lyase. J. Biol. Chem. 245, 2476–2482 (1970).
Davies, G. J., Gloster, T. M., & Henrissat, B. Recent structural insights into the expanding world of carbohydrate-active enzymes. Curr. Opin. Struct. Biol. 15, 637–645 (2005).
Campbell, J. A., Davies, G. J., Bulone, V. & Henrissat, B. A classification of nucleotide-diphospho-sugar glycosyltransferases based on amino acid sequence similarities. Biochem. J. 326, 929–942 (1997).
Coutinho, P. M., Deleury, E., Davies, G. J. & Henrissat, B. An evolving hierarchical family classification for glycosyltransferases. J. Mol. Biol. 328, 307–317 (2003).
Ünligil, U. M. & Rini, J. M. Glycosyltransferase structure and mechanism. Curr. Opin. Struct. Biol. 10, 510–517 (2000).
Bourne, Y. & Henrissat, B. Glycoside hydrolases and glycosyltransferases: families and functional modules. Curr. Opin. Struct. Biol. 11, 593–600 (2001).
Breton, C., Mucha, J. & Jeanneau, C. Structural and functional features of glycosyltransferases. Biochimie 83, 713–718 (2001).
Pak, J. E. et al. X-ray crystal structure of leukocyte type core 2 β1,6-N-acetylglucosaminyltransferase. Evidence for a convergence of metal ion-independent glycosyltransferase mechanism. J. Biol. Chem. 281, 26693–26701 (2006).
Chiu, C. P. et al. Structural analysis of the sialyltransferase CstII from Campylobacter jejuni in complex with a substrate analogue. Nature Struct. Mol. Biol. 11, 163–170 (2004).
Murray, B. W., Takayama, S., Schultz, J. & Wong, C.-H. Mechanism and specificity of human α-1,3-fucosyltransferase V. Biochemistry 35, 11183–11195 (1996).
Hu, Y. & Walker, S. Remarkable structural similarities between diverse glycosyltransferases. Chem. Biol. 9, 1287–1296 (2002).
Sinnott, M. L. Catalytic mechanisms of enzymic glycosyl transfer. Chem. Rev. 90, 1171–1202 (1990).
Gibson, R. P., Turkenburg, J. P., Charnock, S. J., Lloyd, R. & Davies, G. J. Insights into trehalose synthesis provided by the structure of the retaining glucosyltransferase OtsA. Chem. Biol. 9, 1337–1346 (2002).
Pedersen, L. C., Darden, T. A. & Negishi, M. Crystal structure of β1,3-glucuronyltransferase I in complex with active donor substrate UDP-GlcUA. J. Biol. Chem. 277, 21869–21873 (2002).
Charnock, S. J. & Davies, G. J. Structure of the nucleotide-diphospho-sugar transferase, SpsA from Bacillus subtilis, in native and nucleotide-complexed forms. Biochemistry 38, 6380–6385 (1999).
Tarbouriech, N., Charnock, S. J. & Davies, G. J. Three-dimensional structures of the Mn and Mg dTDP complexes of the family GT-2 glycosyltransferase SpsA: a comparison with related NDP-sugar glycosyltransferases. J. Mol. Biol. 314, 655–661 (2001).
Qiao, L., Murray, B. W., Shimazaki, M., Schultz, J. & Wong, C.-H. Synergistic inhibition of human α-1,3-fucosyltransferase V. J. Am. Chem. Soc. 118, 7653–7662 (1996).
Tvaroska, I., Andre, I. & Carver, J. P. Ab initio molecular orbital study of the catalytic mechanism of glycosyltransferases: description of reaction pathways and determination of transition-state structures for inverting N-acetylglucosaminyltransferases. J. Am. Chem. Soc. 122, 8762–8776 (2000).
Lairson, L. L. et al. Intermediate trapping on a mutant retaining α-galactosyltransferase identifies an unexpected aspartate residue. J. Biol. Chem. 279, 28339–28344 (2004).
Zechel, D. L. & Withers, S. G. Glycosidase mechanisms: anatomy of a finely tuned catalyst. Acc. Chem. Res. 33, 11–18 (2000).
Persson, K. et al. Crystal structure of the retaining galactosyltransferase LgtC from Neisseria meningitidis in complex with donor and acceptor sugar analogs. Nature Struct. Biol. 8, 166–175 (2001).
Pedersen, L. C. et al. Crystal structure of an α-1,4-N-acetylhexosaminyltransferase (EXTL2), a member of the exostosin gene family involved in heparan sulfate biosynthesis. J. Biol. Chem. 278, 14420–14428 (2003).
Boix, E. et al. Structure of UDP complex of UDP-galactose:β-galactoside-α-1,3-galactosyltransferase at 1.53-Å resolution reveals a conformational change in the catalytically important C terminus. J. Biol. Chem. 276, 48608–48614 (2001).
Boix, E., Zhang, Y., Swaminathan, G. J., Brew, K. & Acharya, K. R. Structural basis of ordered binding of donor and acceptor substrates to the retaining glycosyltransferase, α-1,3-galactosyltransferase. J. Biol. Chem. 277, 28310–28318 (2002).
Ly, H. D., Lougheed, B., Wakarchuk, W. W. & Withers, S. G. Mechanistic studies of a retaining α-galactosyltransferase from Neisseria meningitidis. Biochemistry 41, 5075–5085 (2002).
Zhang, Y. et al. Roles of individual enzyme-substrate interactions by α-1,3-galactosyltransferase in catalysis and specificity. Biochemistry 42, 13512–13521 (2003).
Borisova, S. A. et al. Substrate specificity of the macrolide-glycosylating enzyme pair DesVII/DesVIII: opportunities, limitations, and mechanistic hypotheses. Angew. Chem. Int. Ed. Engl. 45, 2748–2753 (2006).
Kao, C.-L., Borisova, S. A., Kim, H. J., & Liu, H.-w. Linear aglycones are the substrates for glycosyltransferase DesVII in methymycin biosynthesis: analysis and implications. J. Am. Chem. Soc. 128, 5606–5607 (2006).
Yuan, Y. et al. In vitro reconstitution of EryCIII activity for the preparation of unnatural macrolides. J. Am. Chem. Soc. 127, 14128–14129 (2005).
Lu, W. et al. AknT is an activating protein for the glycosyltransferase AknS in L-aminodeoxysugar transfer to the aglycone of aclacinomycin A. Chem. Biol. 12, 527–534 (2005).
Imerpiali, B. & Tai, V. W. -F. in Carbohydrate-based Drug Discovery Vol. 1 (ed. Wong, C. -H.) 281–303 (Wiley-VCH, Weinheim, 2003).
Onaka, H., Taniguchi, S.-i., Igarashi, Y. & Furumai, T. Characterization of the biosynthetic gene cluster of rebeccamycin from Lechevalieria aerocolonigenes ATCC 39243. Biosci. Biotechnol. Biochem. 67, 127–138 (2003).
Salas, A. P. et al. Deciphering the late steps in the biosynthesis of the anti-tumour indolocarbazole staurosporine: sugar donor substrate flexibility of the StaG glycosyltransferase. Mol. Microbiol. 58, 17–27 (2005).
Gao, Q., Zhang, C., Blanchard, S. & Thorson, J. S. Deciphering indolocarbazole and enediyne aminodideoxypentose biosynthesis through comparative genomics: insights from the AT2433 biosynthetic locus. Chem. Biol. 13, 733–743 (2006).
Wacker, M. et al. N-linked glycosylation in Campylobacter jejuni and its functional transfer into E. coli. Science 298, 1790–1793 (2002).
Hultin, P. G. Bioactive C-glycosides from bacterial secondary metabolism. Curr. Topics Med. Chem. 5, 1299–1331 (2005).
Bililign, T., Hyun, C.-G., Williams, J. S., Czisny, A. M. & Thorson, J. S. The hedamycin locus implicates a novel aromatic PKS priming mechanism. Chem. Biol. 11, 959–969 (2004).
Halkier, B. A. & Gershenzon, J. Biology and biochemistry of glucosinolates. Annu. Rev. Plant Biol. 57, 303–333 (2006).
Grubb, C. D. et al. Arabidopsis glucosyltransferase UGT74B1 functions in glucosinolate biosynthesis and auxin homeostasis. Plant J. 40, 893–908 (2004).
Thiericke, R. & Rohr, J. Biological variation of microbial metabolites by precursor-directed biosynthesis. Nat. Prod. Rep. 10, 265–289 (1993).
Weist, S. & Sussmuth, R. D. Mutational biosynthesis — a tool for the generation of structural diversity in the biosynthesis of antibiotics. Appl. Microbiol. Biotech. 68, 141–150 (2005).
Solenberg, P. J. et al. Production of hybrid glycopeptide antibiotics in vitro and in Streptomyces toyocaensis. Chem. Biol. 4, 195–202 (1997).
Madduri, K. et al. Production of the antitumor drug epirubicin (4′-epidoxorubicin) and its precursor by a genetically engineered strain of Streptomyces peucetius. Nature Biotechnol. 16, 69–74 (1998).
Zhao, L., Sherman, D. H. & Liu, H.-w. Biosynthesis of desosamine: construction of a new methymycin/neomethymycin analogue by deletion of a desoamine biosynthetic gene. J. Am. Chem. Soc. 120, 10256–10257 (1998).
Zhao, L., Que, N. L. S., Xue, Y., Sherman, D. H. & Liu, H.-w. Mechansitic studies of desosamine biosynthesis: C-4 deoxygenation precedes C-3 transamination. J. Am. Chem. Soc. 120, 12159–12160 (1998).
Zhao, L., Borisova, S., Yeung, S. M. & Liu, H. Study of C-4 deoxygenation in the biosynthesis of desosamine: evidence implicating a novel mechanism. J. Am. Chem. Soc. 123, 7909–7910 (2001).
Borisova, S. A., Zhao, L., Sherman, D. H. & Liu, H. W. Biosynthesis of desosamine: construction of a new macrolide carrying a genetically designed sugar moiety. Org. Lett. 1, 133–136 (1999).
Yamase, H., Zhao, L. & Liu, H.-w. Engineering a hybrid sugar biosynthetic pathway: production of L-rhamnose and its implication on dihydrostreptose biosynthesis. J. Am. Chem. Soc. 122, 12397–12398 (2000).
Melancon, C. E., Yu, W. L. & Liu, H. W. TDP-mycaminose biosynthetic pathway revised and conversion of desosamine pathway to mycaminose pathway with one gene. J. Am. Chem. Soc. 127, 12240–12241 (2005).
Tang, L. & McDaniel, R. Construction of desosamine containing polyketide libraries using a glycosyltransferase with broad substrate specificity. Chem. Biol. 8, 547–555 (2001).
Decker, H., Haag, S., Udvarnoki, G. & Rohr, J. Novel genetically engineered tetracenomycins. Angew. Chem. Int. Ed. Engl. 34, 1107–1110 (1995).
Wohlert, S.-E. et al. Novel hybrid tetracenomycins through combinatorial biosynthesis using a glycosyltransferase encoded by the elm genes in cosmid 16F4 and which shows broad sugar substrate specificity. J. Am. Chem. Soc. 120, 10596–10601 (1998).
Rodriguez, L. et al. Engineering deoxysugar biosynthetic pathways from antibiotic-producing microorganisms: a tool to produce novel glycosylated bioactive compounds. Chem. Biol. 9, 721–729 (2002).
Fischer, C. et al. Digitoxosyltetracenomycin C and glucosyltetracenomycin C, two novel elloramycin analogues obtained by exploring the sugar donor substrate flexibility of glycosyltransferase ElmGT. J. Nat. Prod. 65, 1685–1689 (2002).
Lombo, F. et al. Engineering biosynthetic pathways for deoxysugars: branched-chain sugar pathways and derivatives from the antitumor tetracenomycin. Chem. Biol. 11, 1709–1718 (2004).
Perez, M. et al. Combining sugar biosynthesis genes for the generation of L- and D-amicetose and formation of two novel antitumor tetracenomycins. Chem. Commun. (Camb.) 12, 1604–1606 (2005).
Zhang, C. et al. RebG- and RebM-catalyzed indolocarbazole diversification. ChemBioChem 7, 795–804 (2006).
Sanchez, C. et al. Combinatorial biosynthesis of antitumor indolocarbazole compounds. Proc. Natl Acad. Sci. USA 102, 461–466 (2005).
Trefzer, A. et al. Elucidation of the function of two glycosyltransferase genes (lanGT1 and lanGT4) involved in landomycin biosynthesis and generation of new oligosaccharide antibiotics. Chem. Biol. 8, 1239–1252 (2001).
Torkkell, S. et al. The entire nogalamycin biosynthetic gene cluster of Streptomyces nogalater: characterization of a 20-kb DNA region and generation of hybrid structures. Mol. Gen. Genet. 266, 276–288 (2001).
Wohlert, S.-E. et al. Insights about the biosynthesis of the avermectin deoxysugar L-oleandrose through heterologous expression of Streptomyces avermitilis deoxysugar genes in Streptomyces lividans. Chem. Biol. 8, 681–700 (2001).
Gaisser, S. et al. A defined system for hybrid macrolide biosynthesis in Saccharopolyspora erythraea. Mol. Microbiol. 36, 391–401 (2000).
Hoffmeister, D. et al. Engineered urdamycin glycosyltransferases are broadened and altered in substrate specificity. Chem. Biol. 9, 287–295 (2002).
Hoffmeister, D., Ichinose, K. & Bechthold, A. Two sequence elements of glycosyltransferases involved in urdamycin biosynthesis are responsible for substrate specificity and enzymatic activity. Chem. Biol. 8, 557–567 (2001).
Love, K. R., Swoboda, J. G., Noren, C. J. & Walker, S. Enabling glycosyltransferase evolution: a facile substrate-attachment strategy for phage-display enzyme evolution. ChemBioChem 7, 753–756 (2006).
Aharoni, A. et al. High-throughput screening methodology for the directed evolution of glycosyltransferases. Nature Methods 3, 609–614 (2006).
Mulichak, A. M., Losey, H. C., Walsh, C. T. & Garavito, R. M. Structure of the UDP-glucosyltransferase GtfB that modifies the heptapeptide aglycone in the biosynthesis of vancomycin group antibiotics. Structure 9, 547–557 (2001).
Mulichak, A. M. et al. Structure of the TDP-epi-vancosaminyltransferase GtfA from the chloroeremomycin biosynthetic pathway. Proc. Natl Acad. Sci. USA 100, 9238–9243 (2003).
Mulichak, A. M., Lu, W., Losey, H. C., Walsh, C. T. & Garavito, R. M. Crystal structure of vancosaminyltransferase GtfD from the vancomycin biosynthetic pathway: interactions with acceptor and nucleotide ligands. Biochemistry 43, 5170–5180 (2004).
Offen, W. et al. Structure of a flavonoid glucosyltransferase reveals the basis for plant natural product modification. EMBO J. 25, 1396–1405 (2006).
Fu, X. et al. Antibiotic optimization via in vitro glycorandomization. Nature Biotechnol. 21, 1467–1469 (2003).
Albermann, C. et al. Substrate specificity of NovM: implications for novobiocin biosynthesis and glycorandomization. Org. Lett. 5, 933–936 (2003).
Zhang, C. et al. Exploiting the reversibility of natural product glycosyltransferase-catalyzed reactions. Science 313, 1291–1294 (2006).
Minami, A., Kakinuma, K. & Eguchi, T. Aglycon switch approach toward unnatural glycosides from natural glycoside with glycosyltransferase VinC. Tetrahedr. Lett. 46, 6187–6190 (2005).
Melancon, C. E., Thibodeaux, C. J. & Liu, H.-w. Glyco-stripping and glyco-swapping. ACS Chem. Biol. 1, 499–504 (2006).
Grison, C., Petek, S., Finance, C. & Coutrot, P. Synthesis and antibacterial activity of mechanism-based inhibitors of KDO8P synthase and DAH7P synthase. Carbohydrate Res. 340, 529–537 (2005).
Carlson, E. E., May, J. F. & Kiessling, L. L. Chemical probes of UDP-galactopyranose mutase. Chem. Biol. 13, 825–837 (2006).
van Heijenoort, J. Recent advances in the formation of the bacterial peptidoglycan monomer unit. Nat. Prod. Rep. 18, 503–519 (2001).
Zoeiby, A. E., Sanschagrin, F. & Levesque, R. C. Structure and function of the Mur enzymes development of novel inhibitors. Mol. Microbiol. 47, 1–12 (2003).
Lovering, A. L., de Castro, L. H., Lim, D. & Strynadka, N. C. J. Structural insight into the transglycosylation step of bacterial cell-wall biosynthesis. Science 315, 1402–1405 (2007).
Acknowledgements
We thank the National Institutes of Health for their generous support of this work.
Author information
Authors and Affiliations
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Additional information
Note added in proof: Lovering et al. recently demonstrated that the glycosyltransferase domain of a bifunctional glycosyltransferase/transpeptidase enzyme involved in peptidoglycan biosynthesis in Staphylococcus aureus adopts a structural fold that is distinct from that of the GT-A and GT-B glycosyltransferase families, making it the first member of a new family of glycosyltransferases to be structurally characterized98.
Rights and permissions
About this article
Cite this article
Thibodeaux, C., Melançon, C. & Liu, Hw. Unusual sugar biosynthesis and natural product glycodiversification. Nature 446, 1008–1016 (2007). https://doi.org/10.1038/nature05814
Published:
Issue Date:
DOI: https://doi.org/10.1038/nature05814
This article is cited by
-
A conservative distribution of tridomain NDP-heptose synthetases in actinobacteria
Science China Life Sciences (2022)
-
Dual-stimuli-responsive porous polymer enzyme reactor for tuning enzymolysis efficiency
Microchimica Acta (2021)
-
Exploring and applying the substrate promiscuity of a C-glycosyltransferase in the chemo-enzymatic synthesis of bioactive C-glycosides
Nature Communications (2020)
-
Convergent biosynthetic transformations to a bacterial specialized metabolite
Nature Chemical Biology (2019)
-
Isolation and characterization of a multifunctional flavonoid glycosyltransferase from Ornithogalum caudatum with glycosidase activity
Scientific Reports (2018)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.