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. 1993 Sep;59(9):2857–2863. doi: 10.1128/aem.59.9.2857-2863.1993

Construction of a Biotin-Overproducing Strain of Serratia marcescens

Naoki Sakurai 1,, Yuji Imai 1, Makoto Masuda 1, Saburo Komatsubara 1,†,*, Tetsuya Tosa 1
PMCID: PMC182377  PMID: 16349036

Abstract

We have isolated mutants resistant to acidomycin, a biotin analog, from Serratia marcescens Sr41. Strain SB304, resistant to 0.5 mg of acidomycin (frequently called actithiazic acid) per ml, produced 5 mg of d-biotin per liter of a medium containing sucrose and urea. Strain SB412, which was isolated from SB304 on a minimal agar plate containing 2 mg of acidomycin per ml and 0.1 mg of 5-(2-thienyl)-valeric acid per ml, produced 20 mg of d-biotin per ml. The two enzymes related to biotin synthesis were found to be released from biotin-mediated feedback repression in these mutants. Transductional analysis revealed that SB412 had acquired at least two mutations, one in the biotin operon locus and the other in an unknown locus distant from the biotin operon locus.

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Selected References

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  1. Bachmann B. J. Linkage map of Escherichia coli K-12, edition 8. Microbiol Rev. 1990 Jun;54(2):130–197. doi: 10.1128/mr.54.2.130-197.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barker D. F., Campbell A. M. Genetic and biochemical characterization of the birA gene and its product: evidence for a direct role of biotin holoenzyme synthetase in repression of the biotin operon in Escherichia coli. J Mol Biol. 1981 Mar 15;146(4):469–492. doi: 10.1016/0022-2836(81)90043-7. [DOI] [PubMed] [Google Scholar]
  3. Barker D. F., Campbell A. M. The birA gene of Escherichia coli encodes a biotin holoenzyme synthetase. J Mol Biol. 1981 Mar 15;146(4):451–467. doi: 10.1016/0022-2836(81)90042-5. [DOI] [PubMed] [Google Scholar]
  4. Barker D. F., Kuhn J., Campbell A. M. Sequence and properties of operator mutations in the bio operon of Escherichia coli. Gene. 1981 Jan-Feb;13(1):89–102. doi: 10.1016/0378-1119(81)90046-9. [DOI] [PubMed] [Google Scholar]
  5. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  6. Calhoun D. H., Jensen R. A. Significance of altered carbon flow in aromatic amino acid synthesis: an approach to the isolation of regulatory mutants in Pseudomonas aeruginosa. J Bacteriol. 1972 Jan;109(1):365–372. doi: 10.1128/jb.109.1.365-372.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Campbell A., Chang R., Barker D., Ketner G. Biotin regulatory (bir) mutations of Escherichia coli. J Bacteriol. 1980 Jun;142(3):1025–1028. doi: 10.1128/jb.142.3.1025-1028.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Casadaban M. J., Cohen S. N. Lactose genes fused to exogenous promoters in one step using a Mu-lac bacteriophage: in vivo probe for transcriptional control sequences. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4530–4533. doi: 10.1073/pnas.76.9.4530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cleary P. P., Campbell A., Chang R. Location of promoter and operator sites in the biotin gene cluster of Escherichia coli. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2219–2223. doi: 10.1073/pnas.69.8.2219. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. DAVIS B. D., MINGIOLI E. S. Mutants of Escherichia coli requiring methionine or vitamin B12. J Bacteriol. 1950 Jul;60(1):17–28. doi: 10.1128/jb.60.1.17-28.1950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dakshinamurti K., Cheah-Tan C. Biotin-mediated synthesis of hepatic glucokinase in the rat. Arch Biochem Biophys. 1968 Sep 20;127(1):17–21. doi: 10.1016/0003-9861(68)90195-1. [DOI] [PubMed] [Google Scholar]
  12. Das Gupta C. K., Vrancic A., Guha A. Isolation and characterization of the biotin genes of Escherichia coli K-12. Gene. 1977 Jul;1(5-6):331–345. doi: 10.1016/0378-1119(77)90038-5. [DOI] [PubMed] [Google Scholar]
  13. DasGupta C. K., Guha A. Isolation of the regulatory segment of the biotin operons of Escherichia coli K-12. Gene. 1978 May;3(3):233–246. doi: 10.1016/0378-1119(78)90034-3. [DOI] [PubMed] [Google Scholar]
  14. DeMoll E., Shive W. The origin of sulfur in biotin. Biochem Biophys Res Commun. 1983 Jan 14;110(1):243–249. doi: 10.1016/0006-291x(83)91286-x. [DOI] [PubMed] [Google Scholar]
  15. Eisenberg M. A., Hsiung S. C. Mode of action of the biotin antimetabolites actithiazic acid and alpha-methyldethiobiotin. Antimicrob Agents Chemother. 1982 Jan;21(1):5–10. doi: 10.1128/aac.21.1.5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Eisenberg M. A. Mode of action of alpha-dehydrobiotin, a biotin analogue. J Bacteriol. 1975 Jul;123(1):248–254. doi: 10.1128/jb.123.1.248-254.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Eisenberg M. A. Regulation of the biotin operon in E. coli. Ann N Y Acad Sci. 1985;447:335–349. doi: 10.1111/j.1749-6632.1985.tb18449.x. [DOI] [PubMed] [Google Scholar]
  18. Eisenburg M. A., Mee B., Prakash O., Eisenburg M. R. Properties of alpha-dehydrobiotin-resistant mutants of Escherichia coli K-12. J Bacteriol. 1975 Apr;122(1):66–72. doi: 10.1128/jb.122.1.66-72.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gloeckler R., Ohsawa I., Speck D., Ledoux C., Bernard S., Zinsius M., Villeval D., Kisou T., Kamogawa K., Lemoine Y. Cloning and characterization of the Bacillus sphaericus genes controlling the bioconversion of pimelate into dethiobiotin. Gene. 1990 Mar 1;87(1):63–70. doi: 10.1016/0378-1119(90)90496-e. [DOI] [PubMed] [Google Scholar]
  20. Guha A. Divergent orientation of transcription from the biotin locus of Escherichia coli. J Mol Biol. 1971 Feb 28;56(1):53–62. doi: 10.1016/0022-2836(71)90083-0. [DOI] [PubMed] [Google Scholar]
  21. Hanka L. J., Martin D. G., Reineke L. M. Two new antimetabolites of biotin: alpha-methyldethiobiotin and alpha-methylbiotin. Antimicrob Agents Chemother. 1972 Feb;1(2):135–138. doi: 10.1128/aac.1.2.135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hanka L. J., Reineke L. M., Martin D. G. Biological studies with alpha-dehydrobiotin. J Bacteriol. 1969 Oct;100(1):42–46. doi: 10.1128/jb.100.1.42-46.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Iwahara S., McCormick D. B., Wright L. D., Li H. C. Bacterial degradation of biotin. 3. Metabolism of 14C-carbonyl-labeled biotin. J Biol Chem. 1969 Mar 25;244(6):1393–1398. [PubMed] [Google Scholar]
  24. Izumi Y., Morita H., Sato K., Tani Y., Ogata K. Synthesis of biotin-vitamers from pimelic acid and coenzyme A by cell-free extracts of various bacteria. Biochim Biophys Acta. 1972 Mar 30;264(1):210–213. doi: 10.1016/0304-4165(72)90133-x. [DOI] [PubMed] [Google Scholar]
  25. Ketner G., Campbel A. Operator and promoter mutations affecting divergent transcription in the bio gene cluster of Escherichia coli. J Mol Biol. 1975 Jul 25;96(1):13–27. doi: 10.1016/0022-2836(75)90179-5. [DOI] [PubMed] [Google Scholar]
  26. Matsumoto H., Tazaki T., Hosogaya S. A generalized transducing phage of Serratia marcescens. Jpn J Microbiol. 1973 Nov;17(6):473–479. doi: 10.1111/j.1348-0421.1973.tb00933.x. [DOI] [PubMed] [Google Scholar]
  27. Ohsawa I., Speck D., Kisou T., Hayakawa K., Zinsius M., Gloeckler R., Lemoine Y., Kamogawa K. Cloning of the biotin synthetase gene from Bacillus sphaericus and expression in Escherichia coli and Bacilli. Gene. 1989 Aug 1;80(1):39–48. doi: 10.1016/0378-1119(89)90248-5. [DOI] [PubMed] [Google Scholar]
  28. Otsuka A., Abelson J. The regulatory region of the biotin operon in Escherichia coli. Nature. 1978 Dec 14;276(5689):689–694. doi: 10.1038/276689a0. [DOI] [PubMed] [Google Scholar]
  29. Pai C. H. Mutant of Escherichia coli with derepressed levels of the biotin biosynthetic enzymes. J Bacteriol. 1972 Dec;112(3):1280–1287. doi: 10.1128/jb.112.3.1280-1287.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Prakash O., Eisenberg M. A. Biotinyl 5'-adenylate: corepressor role in the regulation of the biotin genes of Escherichia coli K-12. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5592–5595. doi: 10.1073/pnas.76.11.5592. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Reddi A., DeAngelis B., Frank O., Lasker N., Baker H. Biotin supplementation improves glucose and insulin tolerances in genetically diabetic KK mice. Life Sci. 1988;42(13):1323–1330. doi: 10.1016/0024-3205(88)90226-3. [DOI] [PubMed] [Google Scholar]
  32. Sabatié J., Speck D., Reymund J., Hebert C., Caussin L., Weltin D., Gloeckler R., O'Regan M., Bernard S., Ledoux C. Biotin formation by recombinant strains of Escherichia coli: influence of the host physiology. J Biotechnol. 1991 Aug;20(1):29–49. doi: 10.1016/0168-1656(91)90033-r. [DOI] [PubMed] [Google Scholar]
  33. Szybalski E. H., Szybalski W. A physical map of the Escherichia coli bio operon. Gene. 1982 Jul-Aug;19(1):93–103. doi: 10.1016/0378-1119(82)90193-7. [DOI] [PubMed] [Google Scholar]
  34. Takagi T., Kisumi M. Isolation of a versatile Serratia marcescens mutant as a host and molecular cloning of the aspartase gene. J Bacteriol. 1985 Jan;161(1):1–6. doi: 10.1128/jb.161.1.1-6.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Uchida K. M., Otsuka A. J. Isolation and characterization of Escherichia coli birA intragenic suppressors. Mol Gen Genet. 1987 Dec;210(2):234–240. doi: 10.1007/BF00325688. [DOI] [PubMed] [Google Scholar]
  36. Vesely D. L. Biotin enhances guanylate cyclase activity. Science. 1982 Jun 18;216(4552):1329–1330. doi: 10.1126/science.6123152. [DOI] [PubMed] [Google Scholar]
  37. van de Putte P., Cramer S., Giphart-Gassler M. Invertible DNA determines host specificity of bacteriophage mu. Nature. 1980 Jul 17;286(5770):218–222. doi: 10.1038/286218a0. [DOI] [PubMed] [Google Scholar]

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