Skip to main content
Log in

Screening and characterization of Sphingomonas sp. mutant for welan gum biosynthesis at an elevated temperature

  • Original Paper
  • Published:
Bioprocess and Biosystems Engineering Aims and scope Submit manuscript

Abstract

The optimal temperature for the microbial polysaccharide fermentation is no higher than 30 °C, which is economically undesirable due to additional cooling cost. To solve this problem in the case of welan gum production, we obtained the high-temperature-tolerant-producing strain, Sphingomonas sp. HT-1 by atmospheric and room-temperature plasma-induced mutation. Using HT-1, we obtained a concentration and 1 % aqueous viscosity of 26.8 ± 0.34 g/L and 3.50 ± 0.05 Pa s at a comparatively higher optimal temperature (37 °C). HT-1 was further characterized to understand the mechanism by which these properties are improved. Results indicated that high yield could be attributed to the following: (1) enhanced intracellular synthesis, demonstrated by an increase in the activities of key enzymes, and (2) accelerated cross-membrane substrate uptake and product secretion, indicated by improved membrane fluidity and permeability. Temperature tolerance could be attributed to the overexpression of the investigated heat shock proteins and oxidative stress proteins.

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

Access this article

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

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. 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 

  2. Kranenburg R, Boels IC, Kleerebezem M, Vos WM (1999) Genetics and engineering of microbial exopolysaccharides for food: approaches for the production of existing and novel polysaccharides. Curr Opin Biotech 10:498–504

    Article  Google Scholar 

  3. Paris L (2009) Use of thickening agents for producing soft capsules and film production method. US Patent 7612116B2

  4. Li H, Xu H, Xu H, Li S, Ying HJ, Ouyang PK (2011) Enhanced welan gum production using a two-stage agitation speed control process in Alcaligenes sp. CGMCC2428. Bioprocess Biosyst Eng 34:95–102

    Article  CAS  Google Scholar 

  5. Li H, Xu H, Li S, Feng XH, Xu H, Ouyang PK (2011) Effects of dissolved oxygen and shear stress on the synthesis and molecular weight of welan gum produced from Alcaligenes sp. CGMCC2428. Process Biochem 46:1172–1178

    Article  CAS  Google Scholar 

  6. James GO, Hocart CH, Hillier W, Dean Price G, Djordjevic MA (2013) Temperature modulation of fatty acid profiles for biofuel production in nitrogen deprived Chlamydomonas reinhardtii. Bioresour Technol 127:441–447

    Article  CAS  Google Scholar 

  7. Wang LY, Huang ZL, Li G, Zhao HX, Xing XH, Sun WT, Li HP, Gou ZX, Bao CY (2009) Novel mutation breeding method for Streptomyces avermitilis using an atmospheric pressure glow discharge plasma. J Appl Microbiol 108:851–858

    Article  Google Scholar 

  8. Lu Y, Wang LY, Ma K, Li G, Zhang C, Zhao HX, Lai QH, Li HP, Xing XH (2011) Characteristics of hydrogen production of an Enterobacter aerogenes mutant generated by a new atmospheric and room temperature plasma (ARTP). Biochem Eng J 55:17–22

    Article  CAS  Google Scholar 

  9. Guo T, Tang Y, Xi YL, He AY, Sun BJ, Wu H, Liang DF, Jiang M, Ouyang PK (2011) Clostridium beijerinckii mutant obtained by atmospheric pressure glow discharge producing high proportions of butanol and solvent yields. Biotechnol Lett 33:2379–2383

    Article  CAS  Google Scholar 

  10. Laroussi M (2005) Low temperature plasma-based sterilization: overview and state-of-the-art. Plasma Process Polym 2:391–400

    Article  CAS  Google Scholar 

  11. Lin SJ, Wen CY, Wang PM, Huang JC, Wei CL, Chang JW, Chu WS (2010) High-level production of erythritol by mutants of osmophilic Moniliella sp. Process Biochem 45:973–979

    Article  CAS  Google Scholar 

  12. Thorne L, Mikolajczak MJ, Armentrout RW, Pollock TJ (2000) Increasing the yield and viscosity of exopolysaccharides secreted by Sphingomonas by augmentation of chromosomal genes with multiple copies of cloned biosynthetic genes. J Ind Microbiol Biotechnol 25:49–57

    Article  CAS  Google Scholar 

  13. Denich TJ, Beaudette LA, Lee H, Trevors JT (2003) Effect of selected environmental and physico-chemical factors on bacterial cytoplasmic membranes. J Microbiol Methods 52:149–182

    Article  CAS  Google Scholar 

  14. Zhang M, Xiao Y, Zhu RR, Zhang Q, Wang SL (2012) Enhanced thermotolerance and ethanol tolerance in Saccharomyces cerevisiae mutated by high-energy pulse electron beam and protoplast fusion. Bioprocess Biosyst Eng 35:1455–1465

    Article  CAS  Google Scholar 

  15. Wei ZH, Wu H, Bai LQ, Deng ZX, Zhong JJ (2012) Temperature shift-induced reactive oxygen species enhanced validamycin A production in fermentation of Streptomyces hygroscopicus 5008. Bioprocess Biosyst Eng 35:1309–1316

    Article  CAS  Google Scholar 

  16. 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 

  17. 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 

  18. 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 

  19. 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 

  20. 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 

  21. Zhang J, Du GC, Zhang YP, Liao XY, Wang M, Li Y, Chen J (2010) Glutathione protects Lactobacillus sanfranciscensis against freeze-thawing, freeze-drying, and cold treatment. Appl Environ Microbiol 9:2989–2996

    Article  Google Scholar 

  22. Mikhaleva NI, Santini CL, Giordano G, Nesmeyanova MA, Wu LF (1999) Requirement for phospholipids of the translocation of the trimethylamine N-oxide reductase through the Tat pathway in Escherichia coli. FEBS Lett 463:331–335

    Article  CAS  Google Scholar 

  23. Sonesson A, Jantzen E, Bryn K, Tangen T, Eng J, Zahringer U (1994) Composition of 2, 3-dihydroxy fatty acid-containing lipopolysaccharides from Legionella israelensis, Legionella maceachernii and Legionella micdadei. Microbiology 140:1261–1271

    Article  CAS  Google Scholar 

  24. Loh B, Grant C, Hancock REW (1984) Use of the fluorescent probe 1-n-phenylnaphthylamine to study the interactions of aminoglycoside antibiotics with the outer membrane of Pseudomonas aeruginosa. Antimicrob Agents Chemother 26:546–551

    Article  CAS  Google Scholar 

  25. Lehrer R, Barton A, Ganz T (1988) Concurrent assessment of inner and outer membrane permeabilization and bacteriolysis in E. coli by multiple-wavelength spectrophotometry. J Immunol Methods 108:153–158

    Article  CAS  Google Scholar 

  26. Kayali HA, Tarhan L (2004) The effect of glucose and maltose concentrations on pyruvate and ascorbate production, antioxidant enzyme activities and LPO levels in Fusarium equiseti. Process Biochem 39:1519–1524

    Article  Google Scholar 

  27. Hua XF, Wang J, Wu ZJ, Zhang HX, Li HP, Xing XH, Liu Z (2010) A salt tolerant Enterobacter cloacae mutant for bioaugmentation of petroleum and salt-contaminated soil. Biochem Eng J 49:201–206

    Article  CAS  Google Scholar 

  28. Li H, Xu H, Xu H, Li S, Ouyang PK (2010) Biosynthetic pathway of sugar nucleotides essential for welan gum production in Alcaligenes sp. CGMCC2428. Appl Microbiol Biotechnol 86:295–303

    Article  CAS  Google Scholar 

  29. Zhang HT, Zhan XB, Zheng ZY, Wu JR, Nike E, Yu XB, Lin CC (2012) Improved curdlan fermentation process based on optimization of dissolved oxygen combined with pH control and metabolic characterization of Agrobacterium sp. ATCC 31749. Appl Microbiol Biotechnol 93:367–379

    Article  Google Scholar 

  30. Mykytczuk NCS, Trevors JT, Leduc LG, Ferroni GD (2007) Fluorescence polarization in studies of bacterial cytoplasmic membrane fluidity under environmental stress. Prog Biophys Mol Biol 95:60–82

    Article  CAS  Google Scholar 

  31. Rodriguez VS, Sanchez GA, Martinez RJM, Antonio PJ, Randez GF (2007) Fluidization of membrane lipids enhances the tolerance of Saccharomyces cerevisiae to freezing and salt stress. Appl Environ Microbiol 73:110–116

    Article  Google Scholar 

  32. Los DA, Murata N (2004) Membrane fluidity and its roles in the perception of environmental signals. Biochim Biophys Acta 1666:142–157

    Article  CAS  Google Scholar 

  33. Eriksson M, Nielsen PE, Good L (2002) Cell permeabilization and uptake of antisense peptide–peptide nucleic acid (PNA) into Escherichia coli. J Biol Chem 277:7144–7147

    Article  CAS  Google Scholar 

  34. Aguilera A, Peinado RA, Millan C, Ortega JM, Mauicio JC (2006) Relationship between ethanol tolerance, H+-ATPase activity and the lipid composition of the plasma membrane in different wine yeast strains. Int J Food Microbiol 110:34–42

    Article  CAS  Google Scholar 

  35. Lei JJ, Zhao XQ, Ge XM, Bai FW (2007) Ethanol tolerance and the variation of plasma membrane composition of yeast floc populations with different size distribution. J Biotechnol 131:270–275

    Article  CAS  Google Scholar 

  36. Suwannakham S, Yang ST (2005) Enhanced propionic acid fermentation by Propionibacterium acidipropionici mutant obtained by adaptation in a fibrous-bed bioreactor. Biotechnol Bioeng 91:325–327

    Article  CAS  Google Scholar 

  37. Lushchak VI, Bagnyukova TV (2006) Temperature increase results in oxidative stress in goldfish tissues. 1. Indices of oxidative stress. Comp Biochem Phys C 143:30–35

    Google Scholar 

  38. Cao B, Loh KC (2009) Physiological comparison of Pseudomonas putida between two growth phases during cometabolism of 4-chlorophenol in presence of phenol and glutamate: a proteomics approach. J Chem Technol Biotechnol 84:1178–1185

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the National Basic Research Program of China (973) (2013CB733603), the National High Technology Research and Development Program of China (863) (No. 2013AA020301), the National Key Technology R&D Program (2011BAD23B04), the National Nature Science Foundation of China (No. 21106062) (No. 31371732), the Specialized Research Fund for the Doctoral Program of Higher Education (20113221130001), Graduate Student Innovation Project of Jiangsu Province (No. CXZZ13_0463).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Hong Xu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhu, P., Chen, X., Li, S. et al. Screening and characterization of Sphingomonas sp. mutant for welan gum biosynthesis at an elevated temperature. Bioprocess Biosyst Eng 37, 1849–1858 (2014). https://doi.org/10.1007/s00449-014-1159-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00449-014-1159-8

Keywords

Navigation