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Applied Microbiology and Biotechnology

, Volume 89, Issue 3, pp 739–746 | Cite as

Genetic analysis around aminoalcohol dehydrogenase gene of Rhodococcus erythropolis MAK154: a putative GntR transcription factor in transcriptional regulation

  • Nobuyuki Urano
  • Michihiko Kataoka
  • Takeru Ishige
  • Shinji Kita
  • Keiji Sakamoto
  • Sakayu Shimizu
Applied Genetics and Molecular Biotechnology

Abstract

NADP+-dependent aminoalcohol dehydrogenase (AADH) of Rhodococcus erythropolis MAK154 catalyzes the reduction of (S)-1-phenyl-1-keto-2-methylaminopropane ((S)-MAK) to d-pseudoephedrine, which is used as a pharmaceutical. AADH is suggested to participate in aminoalcohol or aminoketone metabolism in this organism because it is induced by the addition of several aminoalcohols, such as 1-amino-2-propanol. Genetic analysis of around the aadh gene showed that some open reading frames (ORFs) are involved in this metabolic pathway. Four of these ORFs might form a carboxysome-like polyhedral organelle, and others are predicted to encode aminotransferase, aldehyde dehydrogenase, phosphotransferase, and regulator protein. OrfE, a homologous ORF of the FadR subfamily of GntR transcriptional regulators, lies downstream from aadh. To investigate whether or not orfE plays a role in the regulation of aadh expression, the gene disruption mutant of R. erythropolis MAK154 was constructed. The ΔorfE strain showed higher AADH activity than wild-type strain. In addition, a transformed strain, which harbored multi-orfE, showed no AADH activity even in the induced condition with 1-amino-2-propanol. These results suggest that OrfE is a negative regulator that represses aadh expression in the absence of 1-amino-2-propanol.

Keywords

Rhodococcus erythropolis Aminoalcohol dehydrogenase Aminoalcohol metabolism GntR family transcriptional regulator 

Notes

Acknowledgements

This work was supported in part by a Grant-in-Aid for Scientific Research, No. 20380051 (to MK), from the Japan Society for the Promotion of Science (JSPS), and by the Targeted Proteins Research Program (TPRP) of the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan.

References

  1. Brinsmade SR, Paldon T, Escalante-Semerena JC (2005) Minimal functions and physiological conditions required for growth of Salmonella enterica on ethanolamine in the absence of the metabolosome. J Bacteriol 187:8039–8046CrossRefGoogle Scholar
  2. Campbell RL, Dekker EE (1973) Formation of D-1-amino-2-propanol from L-threonine by enzymes from Escherichia coli K-12. Biochem Biophys Res Commun 53:432–438CrossRefGoogle Scholar
  3. Campbell RL, Swain RR, Dekker EE (1978) Purification, separation, and characterization of two molecular forms of D-1-amino-2-propanol:NAD+ oxidoreductase activity from extracts of Escherichia coli K-12. J Biol Chem 253:7282–7288Google Scholar
  4. Cheng S, Liu Y, Crowley CS, Yeates TO, Bobik TA (2008) Bacterial microcompartments: their properties and paradoxes. BioEssays 30:1084–1095CrossRefGoogle Scholar
  5. Dekker EE, Swain RR (1968) Formation of Dg-1-amino-2-propanol by a highly purified enzyme from Escherichia coli. Biochim Biophys Acta 158:306–307Google Scholar
  6. Dirusso CC, Black PN (2004) Bacterial long chain fatty acid transport: gateway to a fatty acid-responsive signaling system. J Biol Chem 279:49563–49566CrossRefGoogle Scholar
  7. Faulkner A, Turner JM (1974) Microbial metabolism of amino alcohols. Aminoacetone metabolism via 1-aminopropan-2-ol in Pseudomonas sp. N.C.I.B. 8858. Biochem J 138:263–276Google Scholar
  8. Faust LR, Connor JA, Roof DM, Hoch JA, Babior BM (1990) Cloning, sequencing, and expression of the genes encoding the adenosylcobalamin-dependent ethanolamine ammonia-lyase of Salmonella typhimurium. J Biol Chem 265:12462–12466Google Scholar
  9. Hashimoto Y, Nishiyama M, Yu F, Watanabe I, Horinouchi S, Beppu T (1992) Development of a host-vector system in a Rhodococcus strain and its use for expression of the cloned nitrile hydratase gene cluster. J Gen Microbiol 138:1003–1010Google Scholar
  10. Haydon DJ, Guest JR (1991) A new family of bacterial regulatory proteins. FEMS Microbiol Lett 79:291–296CrossRefGoogle Scholar
  11. Jones A, Turner JM (1971) Microbial metabolism of amino alcohols via aldehydes. J Gen Microbiol 67:379–381Google Scholar
  12. Jones A, Faulkner A, Turner JM (1973) Microbial metabolism of amino alcohols. Metabolism of ethanolamine and 1-aminopropan-2-ol in species of Erwinia and the roles of amino alcohol kinase and amino alcohol O-phosphate phospho-lyase in aldehyde formation. Biochem J 134:959–968Google Scholar
  13. Kataoka M, Nakamura Y, Urano N, Ishige T, Shi G, Kita S, Sakamoto K, Shimizu S (2006) A novel NADP+-dependent L-1-amino-2-propanol dehydrogenase from Rhodococcus erythropolis MAK154: a promising enzyme for the production of double chiral aminoalcohols. Lett Appl Microbiol 43:430–435CrossRefGoogle Scholar
  14. Kataoka M, Ishige T, Urano N, Nakamura Y, Sakuradani E, Fukui S, Kita S, Sakamoto K, Shimizu S (2008) Cloning and expression of the L-1-amino-2-propanol dehydrogenase gene from Rhodococcus erythropolis, and its application to double chiral compound production. Appl Microbiol Biotechnol 80:597–604CrossRefGoogle Scholar
  15. Kelley JJ, Dekker EE (1984) D-1-amino-2-propanol: NAD+ oxidoreductase. Purification and general properties of the large molecular form of the enzyme from Escherichia coli K-12. J Biol Chem 259:2124–2129Google Scholar
  16. Kelley JJ, Dekker EE (1985) Identity of Escherichia coli D-1-amino-2-propanol:NAD+ oxidoreductase with E. coli glycerol dehydrogenase but not with Neisseria gonorrhoeae 1, 2-propanediol:NAD+ oxidoreductase. J Bacteriol 162:170–175Google Scholar
  17. Kerfeld CA, Sawaya MR, Tanaka S, Nguyen CV, Phillips M, Beeby M, Yeates TO (2005) Protein structures forming the shell of primitive bacterial organelles. Science 309:936–938CrossRefGoogle Scholar
  18. Kofoid E, Rappleye C, Stojiljkovic I, Roth J (1999) The 17-gene ethanolamine (eut) operon of Salmonella typhimurium encodes five homologues of carboxysome shell proteins. J Bacteriol 181:5317–5329Google Scholar
  19. Lowe DA, Turner JM (1970) Microbial metabolism of amino ketones: D-1-aminopropan-2-ol and aminoacetone metabolism in Escherichia coli. J Gen Microbiol 63:49–61Google Scholar
  20. Miwa Y, Fujita Y (1988) Purification and characterization of a repressor for the Bacillus subtilis gnt operon. J Biol Chem 263:13252–13257Google Scholar
  21. Pickard MA, Higgins IJ, Turner JM (1968) Purification and properties of L-1-aminopropan-2-ol: NAD oxidoreductase from a pseudomonad grown on DL-1-aminopropan-2-ol. J Gen Microbiol 54:115–126Google Scholar
  22. Rigali S, Derouaux A, Giannotta F, Dusart J (2002) Subdivision of the helix-turn-helix GntR family of bacterial regulators in the FadR, HutC, MocR, and YtrA subfamilies. J Biol Chem 277:12507–12515CrossRefGoogle Scholar
  23. Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, New YorkGoogle Scholar
  24. Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A 74:5463–5467CrossRefGoogle Scholar
  25. Stojiljkovic I, Baumler AJ, Heffron F (1995) Ethanolamine utilization in Salmonella typhimurium: nucleotide sequence, protein expression, and mutational analysis of the cchA cchB eutE eutJ eutG eutH gene cluster. J Bacteriol 177:1357–1366Google Scholar
  26. Turner JM (1967) Microbial metabolism of amino ketones. L-1-aminopropan-2-ol dehydrogenase and L-threonine dehydrogenase in Escherichia coli. Biochem J 104:112–121Google Scholar
  27. Turner JM, Willetts AJ (1967) Amino ketone formation and aminopropanol-dehydrogenase activity in rat-liver preparations. Biochem J 102:511–519Google Scholar
  28. Willetts AJ, Turner JM (1971) Threonine metabolism in a strain of Bacillus subtilis enzymic oxidation of 1-aminopropan-2-ol and aminoacetone. Biochim Biophys Acta 252:98–104Google Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Nobuyuki Urano
    • 1
  • Michihiko Kataoka
    • 1
    • 3
  • Takeru Ishige
    • 1
  • Shinji Kita
    • 2
  • Keiji Sakamoto
    • 2
  • Sakayu Shimizu
    • 1
  1. 1.Division of Applied Life Sciences, Graduate School of AgricultureKyoto UniversityKyotoJapan
  2. 2.Research InstituteDaiichi Fine Chemical Co.TakaokaJapan
  3. 3.Division of Applied Life Sciences, Graduate School of Life and Environmental SciencesOsaka Prefecture UniversitySakaiJapan

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