Skip to main content
SpringerLink
Log in

Biosynthetic pathway of sugar nucleotides essential for welan gum production in Alcaligenes sp. CGMCC2428

  • Applied Microbial and Cell Physiology
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

Welan gum is a microbial polysaccharide produced by Alcaligenes sp. CGMCC2428 that has d-glucose, d-glucuronic acid, d-glucose, and l-rhamnose as the main structural unit. The biosynthetic pathway of sugar nucleotides essential for producing welan gum in this strain was established in the following ways: (1) the detection of the presence of several intermediates and key enzymes; (2) the analysis of the response upon addition of precursors to the culture medium; (3) the correlation of the activities between several key enzymes with the yields of welan gum. With addition of 200-μM glucose-6-phosphate and fructose-6-phosphate, the production of welan gum was improved by 18%. The activities of phosphoglucomutase, phosphomannose isomerase, UDP-glucose pyrophosphorylase, and dTDP-glucose pyrophosphorylase, correlated well with the yields of welan gum. According to these findings, the biosynthetic pathway was proposed to involve the metabolism of glucose via two discrete systems. The first involves conversion of glucose to glucose-6-phosphate, with further reactions producing glucose-1-phosphate and fructose-6-phosphate, which are metabolized to the nucleotide sugar precursors of welan gum. The second system involves metabolism of glucose to synthesize the basic structural skeleton of the cell via central metabolic pathways, including the Entner–Doudoroff pathway, the pentose phosphate pathway, and the tricarboxylic acid cycle.

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
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Arrecubieta C, Garcia E, Lopez R (1996) Demonstration of UDP-glucose dehydrogenase activity in cell extracts of Escherichia coli expressing the pneumococcal cap3A gene required for the synthesis of type 3 capsular polysaccharide. J Bacteriol 178:2971–2974

    CAS  Google Scholar 

  • Aw TY, Jones DP (1982) Direct determination of UDP-glucuronic acid in cell extracts by high-performance liquid chromatography. Anal Biochem 127:32–36

    Article  CAS  Google Scholar 

  • Basu SS, Dotson DD, Raetz CRH (2000) A facile enzymatic synthesis of uridine diphospho-[14C]galacturonic acid. Anal Biochem 280:173–177

    Article  CAS  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 72:248–254

    Article  CAS  Google Scholar 

  • Chandrasekaran R, Radha A, Lee EJ (1994) Structural roles of calcium ions and side chains in welan: an X-ray study. Carbohydr Res 252:183–207

    CAS  Google Scholar 

  • Duine JA, Jongejan JA (1989) Quinoproteins, enzymes with pyrrolo-quinoline quinone as cofactor. Annu Rev Biochem 58:403–426

    Article  CAS  Google Scholar 

  • Escalante A, Wacher RC, Garcoa GM, Farres A (1998) Enzymes involved in carbohydrate metabolism and their role on exopolysaccharide production in Streptococcus thermophilu. J Appl Microbiol 84:108–114

    Article  CAS  Google Scholar 

  • Fialho AM, Moreira LM, Granja AT, Hoffmann K, Popescu A, Sá-Correia I (2007) Biotechnology of the bacterial gellan gum: genes and enzymes of the biosynthetic pathway. In: Pereira MS (ed) A portrait of state-of-the-art research at the technical university of lisbon, 6th edn. Springer, Netherlands, pp 233–250

    Google Scholar 

  • Goosen N, Vermaas DAM, Putte P (1987) Cloning of the genes involved in synthesis of coenzyme pyrrolo-quinoline-quinone from Acinetobacter calcoaceticus. J Bacteriol 169:303–307

    CAS  Google Scholar 

  • Hsu CH, Lo YM (2003) Characterization of xanthan gum biosynthesis in a centrifugal, packed-bed reactor using metabolic flux analysis. Process Biochem 38:1617–1625

    Article  CAS  Google Scholar 

  • Kang KS, Veeder GT, Cottrell IW (1983) Some novel bacterial polysaccharides of recent development. Prog Ind Microbiol 18:231–253

    CAS  Google Scholar 

  • Li M (2006) Effects of some single-gene knockout of the central metobolic enzymes on the metabolism in Escherichia coli. Dissertation, Zhejiang University

  • Li Y, He N, Guan H, Du G, Chen J (2003) A novel polygalacturonic acid bioflocculant REA-11 produced by Corynebacterium glutamicum: a proposed biosynthetic pathway and experimental confirmation. Appl Microbiol Biotechnol 63:200–206

    Article  CAS  Google Scholar 

  • Li S, Xu H, Shi N (2004) Production of a microbial polysaeeharides by fermentation. Food and Fermentation Industries 30:6–9

    Google Scholar 

  • Lin TY, Hassid WZ (1966) Isolation of guanosine diphosphate uronic acids from a marine brown alga, Fucus gardneri Silva. J Biol Chem 241:3283–3293

    CAS  Google Scholar 

  • Martins LO, Sá-Correia I (1991) Gellan gum biosynthetic enzymes in producing and nonproducing variants of Pseudomonas elodea. Biotechnol Appl Biochem 14:357–364

    CAS  Google Scholar 

  • Ng FMW, Dawes EA (1973) Chemostat studies on the regulation of glucose metabolism in Pseudomonas aeruginosa by citrate. Biochem J 132:129–140

    CAS  Google Scholar 

  • Nishioka T, Matsuda K, Fujita Y (2005) Combined analysis of metabolome and transcriptome: catabolism in bacillus subtilis. In: Tomita M (ed) Metabolomics—the frontier of systems biology, 9th edn. Springer, Tokyo, pp 127–140

    Google Scholar 

  • O'Neill MA, Selvendran RR, Morris VJ, Eagles J (1986) Structure of the extracellular polysaccharide produced by the bacterium Alcaligenes (ATCC 31555) species. Carbohydr Res 145:295–313

    Article  Google Scholar 

  • Piggott NH, Sutherland IW, Jarman TR (1981) Enzymes involved in the biosynthesis of alginate by Pseudomonas aeruginosa. Appl Microbiol Biotechnol 13:179–183

    Article  CAS  Google Scholar 

  • Pindar DF, Bucke C (1975) The biosynthesis of alginic acid by Azotobacter vinelandii. Biochem J 152:617–622

    CAS  Google Scholar 

  • Pugashetti BK, Vadas L, Prihar HS, Feingold DS (1983) GDP-mannose dehydrogenase and biosynthesis of alginate-like polysaccharide in a mucoid strain of Pseudomonas aeruginosa. J Bacteriol 153:1107–1110

    CAS  Google Scholar 

  • Rosalam S, England R (2006) Review of xanthan gum production from unmodified starches by Xanthomonas comprestris sp. Enzyme Microb Technol 39:197–207

    Article  CAS  Google Scholar 

  • Sá-Correia I, Darzins A, Wang SK, Berry A, Chakrabarty AM (1987) Alginate biosynthetic enzymes in mucoid and nonmucoid Pseudomonas aeruginosa: overproduction of phosphomannose isomerase, phosphomannomutase, and GDP-mannose pyrophosphorylase by overexpression of the phosphomannose isomerase (pmi) gene. J Bacteriol 169:3224–3231

    Google Scholar 

  • Sá-Correia I, Fialho AM, Videira P, Moreira LM, Marques AR, Albano H (2002) Gellan gum biosynthesis in Sphingomonas paucimobilis ATCC 31461: genes, enzymes and exopolysaccharide production engineering. J Ind Microbiol Biotechnol 29:170–176

    Article  Google Scholar 

  • Shuster CW, Doudoroff M (1967) Purification of 2-keto-3-deoxy-6-phosphohexonate aldolases of Pseudomonas saccharophila. Arch Microbiol 59:279–286

    CAS  Google Scholar 

  • Silva E, Marques AR, Fialho AM, Granja AT, Sá-Correia I (2005) Proteins encoded by Sphingomonas elodea ATCC 31461 rmlA and ugpG genes, involved in gellan gum biosynthesis, exhibit both dTDP- and UDP-glucose pyrophosphorylase activities. Appl Environ Microbiol 71:4703–4712

    Article  CAS  Google Scholar 

  • Tejwani GA, Ramaiah A (1971) Properties of phsophofructokinase from the mucosa of rat jejunum and the relation to the lack of Pasteur effect. Biochem J 125:507–514

    CAS  Google Scholar 

  • Vandamme EJ, Baets SD, Steinbüchel A (2002) Polysaccharides I: polysaccharides from prokaryotes. In: Vandamme EJ (ed) Biopolymers, 5th edn. Wiley-VCH, New York, pp 1–19

    Google Scholar 

  • Videira PA, Cortes LL, Fialho AM, Sá-Correia I (2000) Identification of the pgmG gene, encoding a bifunctional protein with phosphoglucomutase and phosphomannomutase activities, in the gellan gum-producing strain Sphingomonas paucimobilis ATCC 31461. Appl Environ Microbiol 66:2252–2258

    Article  CAS  Google Scholar 

  • Whitfield C, Sutherland IW, Cripps RE (1982) Glucose metabolism in Xanthomonas campestris. J Gen Microbiol 128:981–985

    CAS  Google Scholar 

  • Winkler HH (1973) Distribution of an inducible hexose-phosphate transport system among various bacteria. J Bacteriol 116(2):1079–1081

    CAS  Google Scholar 

  • Zhu G, Li H (2004) Microbiology: science publishing company. Beijing, China

    Google Scholar 

Download references

Acknowledgements

This work was supported by the National Basic Research Program of China (973; 2009CB724700), the National Nature Science Foundation of China (20674038, 20906050), and the Natural Science Foundation of the Jiangsu Higher Education Institutions of China (No. 08KJA180001).

Author information

Authors and Affiliations

  1. State Key Laboratory of Materials-Oriented Chemical Engineering, College of Food Science and Light Industry, Nanjing University of Technology, No. 5 Xinmofan Road, Gulou District, 210009, Nanjing, China

    Hui Li, Hong Xu, Hao Xu, Sha Li & Ping-Kai Ouyang

Authors
  1. Hui Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hong Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hao Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sha Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ping-Kai Ouyang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hong Xu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Li, H., Xu, H., Xu, H. et al. Biosynthetic pathway of sugar nucleotides essential for welan gum production in Alcaligenes sp. CGMCC2428. Appl Microbiol Biotechnol 86, 295–303 (2010). https://doi.org/10.1007/s00253-009-2298-8

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-009-2298-8

Keywords

Search

Navigation