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. 1968 Dec;16(12):1831–1840. doi: 10.1128/am.16.12.1831-1840.1968

Variations in the Growth Response of Four Different Vitamin B12 Assay Microorganisms to the Same Tissue and Standard Preparations1

Elizabeth A Cook 1, Lillian N Ellis 1
PMCID: PMC547780  PMID: 4881959

Abstract

Four microbiological assay procedures (Escherichia coli, Lactobacillus leichmanni, Euglena gracilis, and Ochromonas malhamensis) for the vitamin B12 content of rat liver and kidney tissue were compared. Significant differences in the growth response for portions of the same sample based on the same standard preparations were noted. The liver values with E. coli were strikingly higher than those obtained with the other microorganisms, about which there was rather general agreement. With kidney tissue, E. coli again gave higher values, but not as significantly high as those obtained with L. leichmannii and E. gracilis. O. malhamensis values were significantly lower. These differences were greater than were expected and could not be explained by the differences of specificity and sensitivity of the assay organisms. Several methods of sample preparation, the possible presence of an alkali-heat-stable factor, different methods of sterilization of the sample plus medium, and different chromatographic procedures were explored in an attempt to find explanations for the variations in growth response, with the thought that these various methods would clarify in what form or forms the free and bound vitamin B12 might exist in the samples. No clear-cut explanation was found.

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Selected References

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  1. BAKER H., FRANK O., PASHER I., SOBOTKA H., NATHAN H. A., HUNTER S. H., AARONSON S. Elaboration of vitamin B12 by Escherichia coli. Experientia. 1960 May 15;16:187–187. doi: 10.1007/BF02178977. [DOI] [PubMed] [Google Scholar]
  2. CHIAO J. S., PETERSON W. H. Microbiological assay of vitamin B12 with a mutant strain of Escherichia coli. Appl Microbiol. 1953 Jan;1(1):42–46. doi: 10.1128/am.1.1.42-46.1953. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. FITZGERALD M. E., GRIMSHAW J. J., ROBINSON F. A. The microbiological assay of vitamin B12 in liver extracts. I. Experimental design and problems of validity. J Pharm Pharmacol. 1956 Sep;8(9):635–647. doi: 10.1111/j.2042-7158.1956.tb12195.x. [DOI] [PubMed] [Google Scholar]
  4. FORD J. E., HUTNER S. H. Role of vitamin B12 in the metabolism of microorganisms. Vitam Horm. 1955;(13):101–136. doi: 10.1016/s0083-6729(08)61024-9. [DOI] [PubMed] [Google Scholar]
  5. FORD J. E. The influence of certain derivatives of vitamin B12 upon the growth of micro-organisms. J Gen Microbiol. 1959 Dec;21:693–701. doi: 10.1099/00221287-21-3-693. [DOI] [PubMed] [Google Scholar]
  6. FORD J. E. The microbiological assay of vitamin B12; the specificity of the requirement of Ochromonas malhamensis for cyanocobalamin. Br J Nutr. 1953;7(4):299–306. doi: 10.1079/bjn19530039. [DOI] [PubMed] [Google Scholar]
  7. FOSTER M. A., JONES K. M., WOODS D. D. The purification and properties of a factor containing vitamin B12 concerned in the synthesis of methionine by Escherichia coli. Biochem J. 1961 Sep;80:519–531. doi: 10.1042/bj0800519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. FROST D. V., FRICKE H. H., SPRUTH H. C. Rat growth assay for vitamin B12; correspondence with collaborative microbiological assay results on U.S.P. liver extracts. J Nutr. 1953 Jan 10;49(1):107–120. doi: 10.1093/jn/49.1.107. [DOI] [PubMed] [Google Scholar]
  9. JOHANSSON K. R. Response to an assay of vitamin B12 by a mutant Escherichia coli. Proc Soc Exp Biol Med. 1953 Jul;83(3):448–453. doi: 10.3181/00379727-83-20382. [DOI] [PubMed] [Google Scholar]
  10. KATO T., SHIMIZU S. CHEMICAL STUDIES ON VITAMIN B12 AND ITS RELATED COMPOUNDS. III. ON THE BIOLOGICAL ACTIVITY OF HYDROXOCOBALAMIN FOR OCHROMONAS MALHAMENSIS. J Vitaminol (Kyoto) 1963 Mar 10;9:32–36. doi: 10.5925/jnsv1954.9.32. [DOI] [PubMed] [Google Scholar]
  11. KATO T., SHIMIZU S. Chemical studies on vitamin B-12 and its related compounds. I. Non-cyano type cobamides in fermentation product. J Vitaminol (Kyoto) 1962 Dec 10;8:297–301. doi: 10.5925/jnsv1954.8.297. [DOI] [PubMed] [Google Scholar]
  12. MISTRY S. P., VADOPALAITE I., CHANG I., FIRTH J., JOHNSON B. C. Vitamin B12 and transmethylation in pig, chick, and rat liver homogenates. J Biol Chem. 1955 Feb;212(2):713–722. [PubMed] [Google Scholar]
  13. PICKEN J. C., Jr, BAURIEDEL W. R. Comparative bioautography of vitamin B12 and related growth factors using Euglena gracilis and Lactobacillus leichmannii. Proc Soc Exp Biol Med. 1950 Nov;75(2):511–515. doi: 10.3181/00379727-75-18248. [DOI] [PubMed] [Google Scholar]
  14. PITNEY W. R., BEARD M. F., VAN LOON E. J. The vitamin B12 content of electrophoretic fractions of liver homogenates. J Biol Chem. 1955 Jan;212(1):117–123. [PubMed] [Google Scholar]
  15. SCHEID H. E., SCHWEIGERT B. S. Liberation and microbiological assay of vitamin B12 in animal tissues. J Biol Chem. 1951 Nov;193(1):299–305. [PubMed] [Google Scholar]
  16. Shapiro J. Freezing-Out, a Safe Technique for Concentration of Dilute Solutions. Science. 1961 Jun 30;133(3470):2063–2064. doi: 10.1126/science.133.3470.2063. [DOI] [PubMed] [Google Scholar]
  17. Skeggs H. R. Microbiological assay of vitamin B12. Methods Biochem Anal. 1966;14:53–62. doi: 10.1002/9780470110324.ch2. [DOI] [PubMed] [Google Scholar]
  18. WINSTEN W. A., EIGEN E. Paper chromatography of vitamin B12 and related bacterial growth factors. J Biol Chem. 1949 Nov;181(1):109–120. [PubMed] [Google Scholar]

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