Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1969 Nov;100(2):557–564. doi: 10.1128/jb.100.2.557-564.1969

Characterization of the Biotin Transport System in Saccharomyces cerevisiae1

Thomas O Rogers a, Herman C Lichstein a
PMCID: PMC250127  PMID: 5354931

Abstract

The characteristics of the biotin transport mechanism of Saccharomyces cerevisiae were investigated in nonproliferating cells. Microbiological and radioisotope assays were employed to measure biotin uptake. The vitamin existed intracellularly in both free and bound forms. Free biotin was extracted by boiling water. Chromatography of the free extract showed it to consist entirely of d-biotin. Cellular bound biotin was released by treating cells with 6 n H2SO4. The rate of biotin uptake was linear with time for 10 min, reaching a maximum at about 20 min followed by a gradual loss of accumulated free vitamin from the cells. Biotin was not degraded or converted to vitamers during uptake. Transport was temperature- and pH-dependent, optimum conditions for uptake being 30 C and pH 4.0. Glucose markedly stimulated biotin transport. In its presence, large intracellular free-biotin concentration gradients were established. Iodoacetate inhibited the glucose stimulation of biotin uptake. The rate of vitamin transport increased in a linear fashion with increasing cell mass. The transport system was saturated with increasing concentrations of the vitamin. The apparent Km for uptake was 3.23 × 10−7m. Uptake of radioactive biotin was inhibited by unlabeled biotin and a number of analogues including homobiotin, desthiobiotin, oxybiotin, norbiotin, and biotin sulfone. Proline, hydroxyproline, and 7,8-diaminopelargonic acid did not inhibit uptake. Unlabeled biotin and desthiobiotin exchanged with accumulated intracellular 14C-biotin, whereas hydroxyproline did not.

Full text

PDF
564

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BRITTEN R. J., McCLURE F. T. The amino acid pool in Escherichia coli. Bacteriol Rev. 1962 Sep;26:292–335. doi: 10.1128/br.26.3.292-335.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Birnbaum J., Pai C. H., Lichstein H. C. Biosynthesis of biotin in microorganisms. V. Control of vitamer production. J Bacteriol. 1967 Dec;94(6):1846–1853. doi: 10.1128/jb.94.6.1846-1853.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. CHAN E. C., STEVENSON I. L. On the biotin requirement of Arthrobacter globiformis. Can J Microbiol. 1962 Jun;8:403–405. doi: 10.1139/m62-053. [DOI] [PubMed] [Google Scholar]
  4. COHEN G. N., MONOD J. Bacterial permeases. Bacteriol Rev. 1957 Sep;21(3):169–194. doi: 10.1128/br.21.3.169-194.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chang W. S., Peterson W. H. FACTORS AFFECTING THE BIOTIN CONTENT OF YEASTS. J Bacteriol. 1949 Jul;58(1):33–44. doi: 10.1128/jb.58.1.33-44.1949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dreyfuss J., Pardee A. B. Regulation of sulfate transport in Salmonella typhimurium. J Bacteriol. 1966 Jun;91(6):2275–2280. doi: 10.1128/jb.91.6.2275-2280.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. FERGUSON R. B., LICHSTEIN H. C. Comparison of microorganisms for the assay of bound biotin. J Bacteriol. 1958 Mar;75(3):366–366. doi: 10.1128/jb.75.3.366-366.1958. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. GALE E. F. Assimilation of amino acids by Gram-positive bacteria and some actions of antibiotics thereon. Adv Protein Chem. 1953;8:285–391. doi: 10.1016/s0065-3233(08)60094-7. [DOI] [PubMed] [Google Scholar]
  9. KESSEL D., LUBIN M. Transport of proline in Escherichia coli. Biochim Biophys Acta. 1962 Feb 12;57:32–43. doi: 10.1016/0006-3002(62)91074-0. [DOI] [PubMed] [Google Scholar]
  10. Krueger K. K., Peterson W. H. Metabolism of Biotin and Oxybiotin by Lactobacillus pentosus 124-2. J Bacteriol. 1948 May;55(5):693–703. doi: 10.1128/jb.55.5.693-703.1948. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. LICHSTEIN H. C., FERGUSON R. B. On the permeability of Lactobacillus arabinosus to biotin. J Biol Chem. 1958 Jul;233(1):243–244. [PubMed] [Google Scholar]
  12. LICHSTEIN H. C., WALLER J. R. Factors affecting the accumulation of biotin by Lactobacillus arabinosus. J Bacteriol. 1961 Jan;81:65–69. doi: 10.1128/jb.81.1.65-69.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. MELVILLE D. B., GENGHOF D. S., LEE J. M. Biological properties of biotin d- and l-sulfoxides. J Biol Chem. 1954 Jun;208(2):503–512. [PubMed] [Google Scholar]
  14. OKADA H., HALVORSON H. O. UPTAKE OF ALPHA-THIOETHYL D-GLUCOPYRANOSIDE BY SACCHAROMYCES CEREVISIAE. I. THE GENETIC CONTROL OF FACILITATED DIFFUSION AND ACTIVE TRANSPORT. Biochim Biophys Acta. 1964 Mar 16;82:538–546. doi: 10.1016/0304-4165(64)90445-3. [DOI] [PubMed] [Google Scholar]
  15. PAI C. H., LICHSTEIN H. C. THE BIOSYNTHESIS OF BIOTIN IN MICROORGANISMS. I. THE PHYSIOLOGY OF BIOTIN SYNTHESIS IN ESCHERICHIA COLI. Biochim Biophys Acta. 1965 Apr 12;100:28–35. doi: 10.1016/0304-4165(65)90423-x. [DOI] [PubMed] [Google Scholar]
  16. Rolfe B., Eisenberg M. A. Genetic and biochemical analysis of the biotin loci of Escherichia coli K-12. J Bacteriol. 1968 Aug;96(2):515–524. doi: 10.1128/jb.96.2.515-524.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. SPENCER R. P., BRODY K. R. BIOTIN TRANSPORT BY SMALL INTESTINE OF RAT, HAMSTER, AND OTHER SPECIES. Am J Physiol. 1964 Mar;206:653–657. doi: 10.1152/ajplegacy.1964.206.3.653. [DOI] [PubMed] [Google Scholar]
  18. Tristram H., Neale S. The activity and specificity of the proline permease in wild-type and analogue-resistant strains of Escherichia coli. J Gen Microbiol. 1968 Jan;50(1):121–137. doi: 10.1099/00221287-50-1-121. [DOI] [PubMed] [Google Scholar]
  19. WOOD R. C., HITCHINGS G. H. A study of the uptake and degradation of folic acid, citrovorum factor, aminopterin, and pyrimethamine by bacteria. J Biol Chem. 1959 Sep;234:2381–2385. [PubMed] [Google Scholar]
  20. WRIGHT L. D., CRESSON E. L., DRISCOLL C. A. Bioautography of biotin and certain related compounds. Proc Soc Exp Biol Med. 1954 Jul;86(3):480–483. doi: 10.3181/00379727-86-21140. [DOI] [PubMed] [Google Scholar]
  21. Waller J. R., Lichstein H. C. Biotin transport and accumulation by cells of Lactobacillus plantarum. I. General properties of the system. J Bacteriol. 1965 Oct;90(4):843–852. doi: 10.1128/jb.90.4.843-852.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Waller J. R., Lichstein H. C. Biotin transport and accumulation by cells of Lactobacillus plantarum. II. Kinetics of the system. J Bacteriol. 1965 Oct;90(4):853–856. doi: 10.1128/jb.90.4.853-856.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES