[go: up one dir, main page]
More Web Proxy on the site http://driver.im/ Skip to main content
Springer Nature Link
Log in

Pasteurella multocida sialic acid aldolase: a promising biocatalyst

  • Biotechnologically Relevant Enzymes and Proteins
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

Sialic acid aldolases or N-acetylneuraminate lyases (NanAs) catalyze the reversible aldol cleavage of N-acetylneuraminic acid (Neu5Ac) to form pyruvate and N-acetyl-d-mannosamine (ManNAc). A capillary electrophoresis assay was developed to directly characterize the activities of NanAs in both Neu5Ac cleavage and Neu5Ac synthesis directions. The assay was used to obtain the pH profile and the kinetic data of a NanA cloned from Pasteurella multocida P-1059 (PmNanA) and a previously reported recombinant Escherichia coli K12 NanA (EcNanA). Both enzymes are active in a broad pH range of 6.0–9.0 in both reaction directions and have similar kinetic parameters. Substrates specificity studies showed that 5-O-methyl-ManNAc, a ManNAc derivative, can be used efficiently as a substrate by PmNanA, but not efficiently by EcNanA, for the synthesis of 8-O-methyl Neu5Ac. In addition, PmNanA (250 mg l−1 culture) has a higher expression level (2.5-fold) than EcNanA (94 mg l−1 culture). The higher expression level and a broader substrate tolerance make PmNanA a better catalyst than EcNanA for the chemoenzymatic synthesis of sialic acids and their derivatives.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
£29.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price includes VAT (United Kingdom)

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  • Aisaka K, Uwajima T (1986) Cloning and constitutive expression of the N-acetylneuraminate lyase gene of Escherichia coli. Appl Environ Microbiol 51:562–565

    CAS  PubMed  PubMed Central  Google Scholar 

  • Aisaka K, Igarashi A, Yamaguchi K, Uwajima T (1991) Purification, crystallization and characterization of N-acetylneuraminate lyase from Escherichia coli. Biochem J 276(Pt 2):541–546

    CAS  PubMed  PubMed Central  Google Scholar 

  • Augé C, David S, Gautheron C, Malleron A, Cavaye B (1988) Preparation of six naturally occurring sialic acids with immobilized acylneuraminate pyruvate lyase. New J Chem 12:733–744

    Google Scholar 

  • Augé C, Gautheron C, David S, Malleron A, Cavaye B, Bouxom B (1990) Sialyl aldolase in organic synthesis: from the trout egg acid, 3-deoxy-d-glycero-d-galacto-2-nonulosonic acid (KDN), to branched-chain higher ketoses as possible new chrions. Tetrahedron 46:201–214

    Google Scholar 

  • Barbosa JA, Smith BJ, DeGori R, Ooi HC, Marcuccio SM, Campi EM, Jackson WR, Brossmer R, Sommer M, Lawrence MC (2000) Active site modulation in the N-acetylneuraminate lyase sub-family as revealed by the structure of the inhibitor-complexed Haemophilus influenzae enzyme. J Mol Biol 303:405–421

    CAS  PubMed  Google Scholar 

  • Chen FT, Dobashi TS, Evangelista RA (1998) Quantitative analysis of sugar constituents of glycoproteins by capillary electrophoresis. Glycobiology 8:1045–1052

    CAS  PubMed  Google Scholar 

  • Cheng MC, Lin SL, Wu SH, Inoue S, Inoue Y (1998) High-performance capillary electrophoretic characterization of different types of oligo- and polysialic acid chains. Anal Biochem 260:154–159

    CAS  PubMed  Google Scholar 

  • Comb DG, Roseman S (1960) The sialic acids. I. The structure and enzymatic synthesis of N-acetylneuraminic acid. J Biol Chem 235:2529–2537

    CAS  PubMed  Google Scholar 

  • Deijl CM, Vliegenthart JF (1983) Configuration of substrate and products of N-acetylneuraminate pyruvate-lyase from Clostridium perfringens. Biochem Biophys Res Commun 111:668–674

    CAS  PubMed  Google Scholar 

  • DeVries GH, Binkley SB (1972) N-aetylneuraminic acid aldolase of Clostridium perfringens: purification, properties and mechanism of action. Arch Biochem Biophys 151:234–242

    CAS  PubMed  Google Scholar 

  • Donald SR, Comb G (1962) N-acetylneuraminic acid aldolase. Methods Enzymol 5:391–394

    Google Scholar 

  • Drzeniek R, Scharmann W, Balke E (1972) Neuraminidase and N-acetylneuraminate pyruvate-lyase of Pasteurella multocida. J Gen Microbiol 72:357–368

    CAS  PubMed  Google Scholar 

  • Dufner G, Schworer R, Muller B, Schmidt RR (2000) Base- and sugar-modified cytidine monophosphate N-acetylneuraminic acid (CMP-Neu5Ac) analogues—synthesis and studies with alpha(2-6)-sialyltransferase from rat liver. Eur J Org Chem 8:1467–1482

    Google Scholar 

  • Ferrero MA, Reglero A, Fernandez-Lopez M, Ordas R, Rodriguez-Aparicio LB (1996) N-acetyl-d-neuraminic acid lyase generates the sialic acid for colominic acid biosynthesis in Escherichia coli K1. Biochem J 317(Pt 1):157–165

    CAS  PubMed  PubMed Central  Google Scholar 

  • Hempel G (2003) Biomedical applications of capillary electrophoresis. Clin Chem Lab Med 41:720–723

    CAS  PubMed  Google Scholar 

  • Huang S, Yu H, Chen X (2007) Disaccharides as sialic acid aldolase substrates: synthesis of disaccharides containing a sialic acid at the reducing end. Angew Chem Int Ed Engl 46:2249–2253

    CAS  PubMed  Google Scholar 

  • Izard T, Lawrence MC, Malby RL, Lilley GG, Colman PM (1994) The three-dimensional structure of N-acetylneuraminate lyase from Escherichia coli. Structure 2:361–369

    CAS  PubMed  Google Scholar 

  • Khorlin AY, Privalova IM (1970) Synthesis of N-acetylneuraminic acid 8-methyl ether. Carbohydr Res 13:373–377

    CAS  Google Scholar 

  • Kolisis FN (1986) An immobilized bienzyme system for assay of sialic acid. Biotechnol Appl Biochem 8:148–152

    CAS  PubMed  Google Scholar 

  • Kruger D, Schauer R, Traving C (2001) Characterization and mutagenesis of the recombinant N-acetylneuraminate lyase from Clostridium perfringens: insights into the reaction mechanism. Eur J Biochem 268:3831–3839

    CAS  PubMed  Google Scholar 

  • Lawrence MC, Barbosa JA, Smith BJ, Hall NE, Pilling PA, Ooi HC, Marcuccio SM (1997) Structure and mechanism of a sub-family of enzymes related to N-acetylneuraminate lyase. J Mol Biol 266:381–399

    CAS  PubMed  Google Scholar 

  • Lilley GG, Barbosa JA, Pearce LA (1998) Expression in Escherichia coli of the putative N-acetylneuraminate lyase gene (nanA) from Haemophilus influenzae: overproduction, purification, and crystallization. Protein Expr Purif 12:295–304

    CAS  PubMed  Google Scholar 

  • Meysick KC, Dimock K, Garber GE (1996) Molecular characterization and expression of a N-acetylneuraminate lyase gene from Trichomonas vaginalis. Mol Biochem Parasitol 76:289–292

    CAS  PubMed  Google Scholar 

  • Nees S, Schauer R, Mayer F (1976) Purification and characterization of N-acetylneuraminate lyase from Clostridium perfringens. Hoppe Seylers Z Physiol Chem 357:839–853

    CAS  PubMed  Google Scholar 

  • Ohta Y, Watanabe K, Kimura A (1985) Complete nucleotide sequence of the E. coli N-acetylneuraminate lyase. Nucleic Acids Res 13:8843–8852

    CAS  PubMed  PubMed Central  Google Scholar 

  • Ohta Y, Shimosaka M, Murata K, Tsukada Y, Kimura A (1986) Molecular cloning of the N-acetylneuraminate lyase gene in Escherichia coli K-12. Appl Microbiol Biotechnol 24:386–391

    Google Scholar 

  • Pan Y, Ayani T, Nadas J, Wen S, Guo Z (2004) Accessibility of N-acyl-d-mannosamines to N-acetyl-d-neuraminic acid aldolase. Carbohydr Res 339:2091–2100

    CAS  PubMed  PubMed Central  Google Scholar 

  • Paolo Brunetti AS, Roseman S (1963) Enzymatic determination of sialic acids: N-acylneuraminic acid = N-acyl-d-mannosamine + pyruvate. Methods Enzymol 6:465–473

    Google Scholar 

  • Schauer R (1982) Chemistry, metabolism, and biological functions of sialic acids. Adv Carbohydr Chem Biochem 40:131–234

    CAS  PubMed  Google Scholar 

  • Steenbergen SM, Lichtensteiger CA, Caughlan R, Garfinkle J, Fuller TE, Vimr ER (2005) Sialic acid metabolism and systemic pasteurellosis. Infect Immun 73:1284–1294

    CAS  PubMed  PubMed Central  Google Scholar 

  • Sugahara K, Sugimoto K, Nomura O, Usui T (1980) Enzymatic assay of serum sialic acid. Clin Chim Acta 108:493–498

    CAS  PubMed  Google Scholar 

  • Sun M, Li Y, Chokhawala HA, Henning R, Chen X (2008) N-terminal 112 amino acid residues are not required for the sialyltransferase activity of Photobacterium damsela alpha2,6-sialyltransferase. Biotechnol Lett 30:671–676

    CAS  PubMed  Google Scholar 

  • Traving C, Roggentin P, Schauer R (1997) Cloning, sequencing and expression of the acylneuraminate lyase gene from Clostridium perfringens A99. Glycoconj J 14:821–830

    CAS  PubMed  Google Scholar 

  • Uchida Y, Tsukada Y, Sugimori T (1984) Purification and properties of N-acetylneuraminate lyase from Escherichia coli. J Biochem (Tokyo) 96:507–522

    CAS  Google Scholar 

  • Varki A (1992) Diversity in the sialic acids. Glycobiology 2:25–40

    CAS  PubMed  Google Scholar 

  • Vimr ER, Troy FA (1985) Regulation of sialic acid metabolism in Escherichia coli: role of N-acylneuraminate pyruvate-lyase. J Bacteriol 164:854–860

    CAS  PubMed  PubMed Central  Google Scholar 

  • Watanabe T, Terabe S (2000) Analysis of natural food pigments by capillary electrophoresis. J Chromatogr A 880:311–322

    CAS  PubMed  Google Scholar 

  • Wong CH, Whitesides GM (1994) Enzymes in synthetic organic chemistry. Elsevier, Oxford, pp 215–228

    Google Scholar 

  • Yu H, Chen X (2006) Aldolase-catalyzed synthesis of beta-d-galp-(1->9)-d-KDN: a novel acceptor for sialyltransferases. Org Lett 8:2393–2396

    CAS  PubMed  Google Scholar 

  • Yu H, Yu H, Karpel R, Chen X (2004) Chemoenzymatic synthesis of CMP-sialic acid derivatives by a one-pot two-enzyme system: comparison of substrate flexibility of three microbial CMP-sialic acid synthetases. Bioorg Med Chem 12:6427–6435

    CAS  PubMed  Google Scholar 

  • Yu H, Chokhawala H, Karpel R, Yu H, Wu B, Zhang J, Zhang Y, Jia Q, Chen X (2005) A multifunctional Pasteurella multocida sialyltransferase: a powerful tool for the synthesis of sialoside libraries. J Am Chem Soc 127:17618–17619

    CAS  PubMed  Google Scholar 

  • Yu H, Huang S, Chokhawala H, Sun M, Zheng H, Chen X (2006) Highly efficient chemoenzymatic synthesis of naturally occurring and non-natural alpha-2,6-linked sialosides: a P. damsela alpha-2,6-sialyltransferase with extremely flexible donor-substrate specificity. Angew Chem Int Ed Engl 45:3938–3944

    CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

This work was supported by grant R01GM076360 from the National Institutes of Health.

Author information

Authors and Affiliations

  1. Department of Chemistry, University of California, One Shields Avenue, Davis, CA, 95616, USA

    Yanhong Li, Hai Yu, Hongzhi Cao, Kam Lau, Saddam Muthana, Vinod Kumar Tiwari, Bryan Son & Xi Chen

Authors
  1. Yanhong Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hai Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hongzhi Cao
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kam Lau
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Saddam Muthana
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Vinod Kumar Tiwari
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Bryan Son
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Xi Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Xi Chen.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(PDF 52.5 KB)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Li, Y., Yu, H., Cao, H. et al. Pasteurella multocida sialic acid aldolase: a promising biocatalyst. Appl Microbiol Biotechnol 79, 963–970 (2008). https://doi.org/10.1007/s00253-008-1506-2

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-008-1506-2

Keywords

Search

Navigation