Skip to main content
Article thumbnail
Location of Repository

Proteomic Analysis of Ketogulonicigenium vulgare under Glutathione Reveals High Demand for Thiamin Transport and Antioxidant Protection

By Qian Ma, Weiwen Zhang, Lu Zhang, Bin Qiao, Chensong Pan, Hong Yi, Lili Wang and Ying-jin Yuan


Ketogulonicigenium vulgare, though grows poorly when mono-cultured, has been widely used in the industrial production of the precursor of vitamin C with the coculture of Bacillus megaterium. Various efforts have been made to clarify the synergic pattern of this artificial microbial community and to improve the growth and production ability of K. vulgare, but there is still no sound explanation. In previous research, we found that the addition of reduced glutathione into K. vulgare monoculture could significantly improve its growth and productivity. By performing SEM and TEM, we observed that after adding GSH into K. vulgare monoculture, cells became about 4–6 folds elongated, and formed intracytoplasmic membranes (ICM). To explore the molecular mechanism and provide insights into the investigation of the synergic pattern of the co-culture system, we conducted a comparative iTRAQ-2-D-LC-MS/MS-based proteomic analysis of K. vulgare grown under reduced glutathione. Principal component analysis of proteomic data showed that after the addition of glutathione, proteins for thiamin/thiamin pyrophosphate (TPP) transport, glutathione transport and the maintenance of membrane integrity, together with several membrane-bound dehydrogenases had significant up-regulation. Besides, several proteins participating in the pentose phosphate pathway and tricarboxylic acid cycle were also up-regulated. Additionally, proteins combating intracellular reactive oxygen species were also up-regulated, which similarly occurred in K. vulgare when the co-cultured B. megaterium cells lysed from our former research results. This study reveals the demand for transmembrane transport of substrates, especially thiamin, and the demand for antioxidant protection of K. vulgare

Topics: Research Article
Publisher: Public Library of Science
OAI identifier:
Provided by: PubMed Central
Download PDF:
Sorry, we are unable to provide the full text but you may find it at the following location(s):
  • http://www.pubmedcentral.nih.g... (external link)
  • Suggested articles


    1. (2009). A method of increased production of 2-keto-L-gulonic acid by Gluconobacter oxydans. China Patent CN200910069697.7.
    2. (2003). A novel L-sorbose dehydrogenase gene and the encoding protein. China Patent CN03102060.7.
    3. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
    4. (2002). Cellular glutathione and thiols metabolism.
    5. (1976). Change in quantity of lipids and cell size during intracytoplasmic membrane formation in Gluconobacter oxydans.
    6. (2009). Comparative proteomic analysis of tolerance and adaptation of ethanologenic Saccharomyces cerevisiae to furfural, a lignocellulosic inhibitory compound.
    7. (2005). Complex cellular responses to reactive oxygen species.
    8. (2010). Continuous 2-keto-L-gulonic acid fermentation by mixed culture of Ketogulonicigenium vulgare DSM 4025 and Bacillus megaterium or Xanthomonas maltophilia.
    9. (2005). Data mining of the transcriptome of Plasmodium falciparum: the pentose phosphate pathway and ancillary processes.
    10. (2011). Development of chemically defined media supporting high cell density growth of Ketogulonicigenium vulgare and Bacillus megaterium.
    11. (1982). Effect of intracytoplasmic membrane development on oxidation of sorbitol and other polyols by Gluconobacter oxydans.
    12. (2000). Effects of menadione and hydrogen peroxide on glutathione status in growing Escherichia coli.
    13. (1982). Energy metabolism in the lateral vestibular nucleus in pyrithiamin-induced thiamin deficiency.
    14. (2001). Energy-dependent conformational change in the TolA protein of Escherichia coli involves its Nterminal domain, TolQ, and TolR.
    15. (1990). Fermentation process. United States Patent 4935359.
    16. (2004). Folate requirements of the 2-keto-L-gulonic acid-producing strain Ketogulonigenium vulgare LMP P-20356 in L-sorbose/CSL medium.
    17. (1989). Free radicals in biology and medicine,
    18. (1993). Genetic and biochemical characterization of the oligopeptide transport system of Lactococcus lactis.
    19. (2003). Glutathione protects Lactococcus lactis against oxidative stress.
    20. (2007). Identification of promoters of two dehydrogenase genes in Ketogulonicigenium vulgare DSM 4025 and their strength comparison in K. vulgare and Escherichia coli.
    21. (2011). Integrated proteomic and metabolomic analysis of an artificial microbial community for two-step production of vitamin C.
    22. (1975). Intracytoplasmic membrane formation and increased oxidation of glycerol during growth of Gluconobacter oxydans.
    23. (2003). Isolation and characterization of mutants of the Bacilllus subtilis oligopeptide permease with altered specificity of oligopeptide transport.
    24. (1993). Membrane topology of the Escherichia coli TolR protein required for cell envelope integrity.
    25. (2011). Metabolome profiling reveals metabolic cooperation between Bacillus megaterium and Ketogulonigenium vulgare during induced swarm motility. Appl Environ Microbiol;doi: 10.1128/ AEM.05123–11.
    26. (1990). Microbiol processes for ascorbic acid biosynthesis: a review.
    27. (1928). Observations on the functions of peroxidase systems and the chemistry of the adrenal cortex.
    28. (2001). Oxidative stress and mechanisms of protection against it in bacteria.
    29. (2010). Past, present, and future industrial biotechnology in China.
    30. (1994). Pharmacology of vitamin C.
    31. (1974). Protein synthesis and breakdown in the mother-cell and forespore compartments during spore morphogenesis in Bacillus megaterium.
    32. (2009). Proteomic insights into adaptive responses of Saccharomyces cerevisiae to the repeated vacuum fermentation.
    33. (2006). Pyrroloquinoline quinone-dependent dehydrogenases from Ketogulonicigenium vulgare catalyze the direct conversion of L-sorbosone to L-ascorbic acid.
    34. (2009). Recent patents on vitamin C: opportunities for crop improvement and single-step biological manufacture. Recent Pat Food Nutr Agric
    35. (1981). Scanning electron microscope evidence for bacterial colonization of a drinking-water distribution system.
    36. (1997). Shainberg A
    37. (2001). Taxonomic characterization of Ketogulonigenium vulgare gen. nov., sp. nov. and Ketogulonigenium robustum sp. nov., which oxidize L-sorbose to 2-keto-L-gulonic acid.
    38. (2000). The COG database: a tool for genome-scale analysis of protein functions and evolution.
    39. (2001). The COG database: new developments in phylogenetic classification of proteins from complete genomes.
    40. (1997). The role of the thioredoxin and glutaredoxin pathways in reducing protein disulfide bonds in the Escherichia coli cytoplasm.
    41. (2006). The use of microorganisms in L-ascorbic acid production.
    42. (2002). Thiamine derivatives bindmessenger RNAs directly to regulate bacterial gene expression.
    43. (2005). Thiamine pyrophosphate riboswitches rre targets for the antimicrobial compound pyrithiamine.
    44. (1998). thiBPQ encodes an ABC transporter required for transport of thiamine and thiamine pyrophosphate in Salmonella typhimurium.
    45. (1966). Trotta A
    46. (2002). Vianney A

    To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.