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.
Similar content being viewed by others
References
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
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
Paris L (2009) Use of thickening agents for producing soft capsules and film production method. US Patent 7612116B2
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
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
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
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
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
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
Laroussi M (2005) Low temperature plasma-based sterilization: overview and state-of-the-art. Plasma Process Polym 2:391–400
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
Los DA, Murata N (2004) Membrane fluidity and its roles in the perception of environmental signals. Biochim Biophys Acta 1666:142–157
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
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
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
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
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
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
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
Corresponding author
Rights 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
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00449-014-1159-8