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. 1994 May;60(5):1495–1499. doi: 10.1128/aem.60.5.1495-1499.1994

Vitamin B12 production by Citrobacter freundii or Klebsiella pneumoniae during tempeh fermentation and proof of enterotoxin absence by PCR.

S Keuth 1, B Bisping 1
PMCID: PMC201508  PMID: 8017933

Abstract

The influence of some fermentation parameters on vitamin B12 formation by strains of Citrobacter freundii and Klebsiella pneumoniae isolated from Indonesian tempeh samples during tempeh fermentation was investigated. A decrease in fermentation temperature from 32 to 24 degrees C led to a decrease in vitamin B12 formation. Inoculation of soybeans with different numbers of cells of C. freundii at the beginning of solid-substrate fermentation showed that only the velocity of vitamin formation and not the final amount of vitamin formed depended on the number of cells. The addition of cobalt and 5,6-dimethylbenzimidazole increased the vitamin B12 content of tempeh. Nevertheless, levels of incorporation of the two precursors into the vitamin B12 molecule were very low. Neither C. freundii nor K. pneumoniae possessed the genes encoding the enterotoxins Shiga-like toxin SLT IIA, heat-labile enterotoxin LT Ih, and heat-stable enterotoxin ST Ih, as indicated by PCR. This result supports the suggested use of these two strains to form vitamin B12 during tempeh fermentation in Indonesia.

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Selected References

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  1. Betley M. J., Miller V. L., Mekalanos J. J. Genetics of bacterial enterotoxins. Annu Rev Microbiol. 1986;40:577–605. doi: 10.1146/annurev.mi.40.100186.003045. [DOI] [PubMed] [Google Scholar]
  2. Bisping B., Hering L., Baumann U., Denter J., Keuth S., Rehm H. J. Tempe fermentation: some aspects of formation of gamma-linolenic acid, proteases and vitamins. Biotechnol Adv. 1993;11(3):481–493. doi: 10.1016/0734-9750(93)90016-g. [DOI] [PubMed] [Google Scholar]
  3. Drelichman V., Band J. D. Bacteremias due to Citrobacter diversus and Citrobacter freundii. Incidence, risk factors, and clinical outcome. Arch Intern Med. 1985 Oct;145(10):1808–1810. [PubMed] [Google Scholar]
  4. Echeverria P., Taylor D. N., Seriwatana J., Chatkaeomorakot A., Khungvalert V., Sakuldaipeara T., Smith R. D. A comparative study of enterotoxin gene probes and tests for toxin production to detect enterotoxigenic Escherichia coli. J Infect Dis. 1986 Feb;153(2):255–260. doi: 10.1093/infdis/153.2.255. [DOI] [PubMed] [Google Scholar]
  5. Friedmann H. C., Cagen L. M. Microbial biosynthesis of B12-like compounds. Annu Rev Microbiol. 1970;24:159–208. doi: 10.1146/annurev.mi.24.100170.001111. [DOI] [PubMed] [Google Scholar]
  6. Guarino A., Giannella R., Thompson M. R. Citrobacter freundii produces an 18-amino-acid heat-stable enterotoxin identical to the 18-amino-acid Escherichia coli heat-stable enterotoxin (ST Ia). Infect Immun. 1989 Feb;57(2):649–652. doi: 10.1128/iai.57.2.649-652.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gunzer F., Böhm H., Rüssmann H., Bitzan M., Aleksic S., Karch H. Molecular detection of sorbitol-fermenting Escherichia coli O157 in patients with hemolytic-uremic syndrome. J Clin Microbiol. 1992 Jul;30(7):1807–1810. doi: 10.1128/jcm.30.7.1807-1810.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gyles C. L. Escherichia coli cytotoxins and enterotoxins. Can J Microbiol. 1992 Jul;38(7):734–746. doi: 10.1139/m92-120. [DOI] [PubMed] [Google Scholar]
  9. Herbert V. Vitamin B-12: plant sources, requirements, and assay. Am J Clin Nutr. 1988 Sep;48(3 Suppl):852–858. doi: 10.1093/ajcn/48.3.852. [DOI] [PubMed] [Google Scholar]
  10. Keuth S., Bisping B. Formation of vitamins by pure cultures of tempe moulds and bacteria during the tempe solid substrate fermentation. J Appl Bacteriol. 1993 Nov;75(5):427–434. doi: 10.1111/j.1365-2672.1993.tb02798.x. [DOI] [PubMed] [Google Scholar]
  11. Liem I. T., Steinkraus K. H., Cronk T. C. Production of vitamin B-12 in tempeh, a fermented soybean food. Appl Environ Microbiol. 1977 Dec;34(6):773–776. doi: 10.1128/aem.34.6.773-776.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Meyer T., Karch H. Genes coding for Shiga-like toxin and heat-stabile enterotoxin in porcine strains of Escherichia coli. FEMS Microbiol Lett. 1989 Mar;49(1):115–119. doi: 10.1016/0378-1097(89)90353-4. [DOI] [PubMed] [Google Scholar]
  13. Moseley S. L., Echeverria P., Seriwatana J., Tirapat C., Chaicumpa W., Sakuldaipeara T., Falkow S. Identification of enterotoxigenic Escherichia coli by colony hybridization using three enterotoxin gene probes. J Infect Dis. 1982 Jun;145(6):863–869. doi: 10.1093/infdis/145.6.863. [DOI] [PubMed] [Google Scholar]
  14. Moseley S. L., Hardy J. W., Hug M. I., Echeverria P., Falkow S. Isolation and nucleotide sequence determination of a gene encoding a heat-stable enterotoxin of Escherichia coli. Infect Immun. 1983 Mar;39(3):1167–1174. doi: 10.1128/iai.39.3.1167-1174.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mulyowidarso R. K., Fleet G. H., Buckle K. A. Association of bacteria with the fungal fermentation of soybean tempe. J Appl Bacteriol. 1990 Jan;68(1):43–47. doi: 10.1111/j.1365-2672.1990.tb02546.x. [DOI] [PubMed] [Google Scholar]
  16. Mulyowidarso R. K., Fleet G. H., Buckle K. A. The microbial ecology of soybean soaking for tempe production. Int J Food Microbiol. 1989 Feb;8(1):35–46. doi: 10.1016/0168-1605(89)90078-0. [DOI] [PubMed] [Google Scholar]
  17. Schmidt H., Montag M., Bockemühl J., Heesemann J., Karch H. Shiga-like toxin II-related cytotoxins in Citrobacter freundii strains from humans and beef samples. Infect Immun. 1993 Feb;61(2):534–543. doi: 10.1128/iai.61.2.534-543.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Seriwatana J., Echeverria P., Escamilla J., Glass R., Huq I., Rockhill R., Stoll B. J. Identification of enterotoxigenic Escherichia coli in patients with diarrhea in Asia with three enterotoxin gene probes. Infect Immun. 1983 Oct;42(1):152–155. doi: 10.1128/iai.42.1.152-155.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Yamamoto T., Yokota T. Sequence of heat-labile enterotoxin of Escherichia coli pathogenic for humans. J Bacteriol. 1983 Aug;155(2):728–733. doi: 10.1128/jb.155.2.728-733.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]

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