Skip to main content
SpringerLink
Log in

Enzymatic production of α-d-galactose 1-phosphate by lactose phosphorolysis

  • Original Research Paper
  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

α-d-Galactose 1-phosphate (αGal1P) is an important building block for the synthesis of nucleotide sugars that are substrates for glycosyltransferases. We have previously reported the creation of novel lactose phosphorylase enzymes that are useful for the synthesis of αGal1P from the cheap and abundant lactose. Here we describe the application of such a lactose phosphorylase in a production system using permeabilized Escherichia coli cells. After purification of the product from the reaction mixture by anion-exchange chromatography and ethanol precipitation, 9.5 grams of highly pure αGal1P were obtained from a 1 l reaction volume.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, Baba M, Datsenko KA, Tomita M, Wanner BL, Mori H (2006) Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection. Mol Syst Biol 2:1–11

    Article  Google Scholar 

  • Bolton PG, Dean ACR (1972) Phosphatase synthesis in Klebsiella (Aerobacter) aerogenes growing in continuous culture. Biochem J 127:87–96

    CAS  PubMed  Google Scholar 

  • Bülter T, Elling L (1999) Enzymatic synthesis of nucleotide sugars. Glycoconj J 16:147–159

    Article  PubMed  Google Scholar 

  • Canovas M, Torroglosa T, Iborra JL (2005) Permeabilization of Escherichia coli cells in the biotransformation of trimethylammonium compounds into l-carnitine. Enzym Microb Technol 37:300–308

    Article  CAS  Google Scholar 

  • Chen X, Liu ZY, Zhang JB, Zhang W, Kowal P, Wang PG (2002) Reassembled biosynthetic pathway for large-scale carbohydrate synthesis: alpha-Gal epitope producing “superbug”. ChemBioChem 3:47–53

    Article  CAS  PubMed  Google Scholar 

  • De Groeve MRM, De Baere M, Hoflack L, Desmet T, Vandamme EJ, Soetaert W (2009) Creating lactose phosphorylase enzymes by directed evolution of cellobiose phosphorylase. Protein Eng Des Sel 22:393–399

    Article  PubMed  Google Scholar 

  • Gawronski JD, Benson DR (2004) Microtiter assay for glutamine synthetase biosynthetic activity using inorganic phosphate detection. Anal Biochem 327:114–118

    Article  CAS  PubMed  Google Scholar 

  • Goedl C, Schwarz A, Minani A, Nidetzky B (2007) Recombinant sucrose phosphorylase from Leuconostoc mesenteroides: characterization, kinetic studies of transglucosylation, and application of immobilised enzyme for production of alpha-d-glucose 1-phosphate. J Biotechnol 129:77–86

    Article  CAS  PubMed  Google Scholar 

  • Hoffmeister D, Yang J, Liu L, Thorson JS (2003) Creation of the first anomeric d/l-sugar kinase by means of directed evolution. Proc Natl Acad Sci USA 100:13184–13189

    Article  CAS  PubMed  Google Scholar 

  • Hokse H (1983) Purification of alpha-d-glucose-1-phosphate. Starke 35:101–102

    Article  CAS  Google Scholar 

  • Nidetzky B, Eis C, Albert M (2000) Role of non-covalent enzyme–substrate interactions in the reaction catalysed by cellobiose phosphorylase from Cellulomonas uda. Biochem J 351:649–659

    Article  CAS  PubMed  Google Scholar 

  • Pham TT, Shah NP (2007) Biotransformation of isoflavone glycosides by Bifidobacterium animalis soymilk supplemented with skim milk powder. J Food Sci 72:M316–M324

    Article  CAS  PubMed  Google Scholar 

  • Posternak T (1950) Synthesis of alpha-d-glucose-1-phosphate and alpha-d-galactose-1-phosphate. J Am Chem Soc 72:4824–4825

    Article  CAS  Google Scholar 

  • Pradel E, Boquet PL (1989) Mapping of the Escherichia coli acid glucose-1-phosphatase gene agp and analysis of its expression in vivo by use of an agp-phoA protein fusion. J Bacteriol 171:3511–3517

    CAS  PubMed  Google Scholar 

  • Rossolini GM, Thaller MC, Pezzi R, Satta G (1994) Identification of an Escherichia coli periplasmic acid phosphatase containing of a 27 kDa polypeptide component. FEMS Microbiol Lett 118:167–173

    Article  CAS  PubMed  Google Scholar 

  • Zhang J, Witholt B, Li Z (2006) Coupling of permeabilized microorganisms for efficient enantioselective reduction of ketone with cofactor recycling. Chem Commun 39:8–400

    Google Scholar 

Download references

Acknowledgements

Research funded by a Ph.D grant (SB51293, SB53293) of the Institute for the Promotion of Innovation through Science and Technology in Flanders (IWT-Vlaanderen).

Author information

Authors and Affiliations

  1. Laboratory of Industrial Microbiology and Biocatalysis, Department of Biochemical and Microbial Technology, Ghent University, Coupure Links 653, 9000, Ghent, Belgium

    Manu R. M. De Groeve, Veerle Depreitere, Tom Desmet & Wim Soetaert

Authors
  1. Manu R. M. De Groeve
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Veerle Depreitere
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Tom Desmet
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Wim Soetaert
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Manu R. M. De Groeve.

Rights and permissions

Reprints and permissions

About this article

Cite this article

De Groeve, M.R.M., Depreitere, V., Desmet, T. et al. Enzymatic production of α-d-galactose 1-phosphate by lactose phosphorolysis. Biotechnol Lett 31, 1873–1877 (2009). https://doi.org/10.1007/s10529-009-0087-1

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10529-009-0087-1

Keywords

Search

Navigation