HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Louis, P. Articles by Galinski, E. A. Search for Related Content PubMed PubMed Citation Articles by Louis, P. Articles by Galinski, E. A. Agricola Articles by Louis, P. Articles by Galinski, E. A.

Characterization of genes for the biosynthesis of the compatible solute ectoine from Marinococcus halophilus and osmoregulated expression in Escherichia coli

Institut für Mikrobiologie & Biotechnologie, Rheinische Friedrich-Wilhelms-Universität Bonn, Meckenheimer Allee 168, 53115 Bonn, Germany

Author for correspondence: Erwin A. Galinski. Tel: + 49 228 733799. Fax: +49 228 737576.

ABSTRACT

The genes of the biosynthetic pathway of ectoine (1,4,5,6-tetrahydro-2-methyl-4-pyrimidinecarboxylic acid) from the Gram-positive moderate halophile Marinococcus halophilus were cloned by functional expression in Escherichia coli. These genes were not only expressed, but also osmoregulated in E. coli, as demonstrated by increasing cytoplasmic ectoine concentration in response to medium salinity. Sequencing of a 4.4 kb fragment revealed four major ORFs, which were designated ectA, ectB, ectC and orfA. The significance of three of these genes for ectoine synthesis was proved by sequence comparison with known proteins and by physiological experiments. Several deletion derivatives of the sequenced fragment were introduced into E. coli and the resulting clones were investigated for their ability to synthesize ectoine or one of the intermediates in its biosynthetic pathway. It was demonstrated that ectA codes for L-2,4-diaminobutyric acid acetyltransferase, ectB for L-2,4-diaminobutyric acid transaminase and ectC for L-ectoine synthase. A DNA region upstream of ectA was shown to be necessary for the regulated expression of ectoine synthesis in response to the osmolarity of the medium.

This article has been cited by other articles:

				L. M. Naughton, S. L. Blumerman, M. Carlberg, and E. F. Boyd Osmoadaptation among Vibrio Species and Unique Genomic Features and Physiological Responses of Vibrio parahaemolyticus Appl. Envir. Microbiol., May 1, 2009; 75(9): 2802 - 2810. [Abstract] [Full Text] [PDF]

				J. Bursy, A. U. Kuhlmann, M. Pittelkow, H. Hartmann, M. Jebbar, A. J. Pierik, and E. Bremer Synthesis and Uptake of the Compatible Solutes Ectoine and 5-Hydroxyectoine by Streptomyces coelicolor A3(2) in Response to Salt and Heat Stresses Appl. Envir. Microbiol., December 1, 2008; 74(23): 7286 - 7296. [Abstract] [Full Text] [PDF]

				A. U. Kuhlmann, J. Bursy, S. Gimpel, T. Hoffmann, and E. Bremer Synthesis of the Compatible Solute Ectoine in Virgibacillus pantothenticus Is Triggered by High Salinity and Low Growth Temperature Appl. Envir. Microbiol., July 15, 2008; 74(14): 4560 - 4563. [Abstract] [Full Text] [PDF]

				J. Bursy, A. J. Pierik, N. Pica, and E. Bremer Osmotically Induced Synthesis of the Compatible Solute Hydroxyectoine Is Mediated by an Evolutionarily Conserved Ectoine Hydroxylase J. Biol. Chem., October 26, 2007; 282(43): 31147 - 31155. [Abstract] [Full Text] [PDF]

				T. Schubert, T. Maskow, D. Benndorf, H. Harms, and U. Breuer Continuous Synthesis and Excretion of the Compatible Solute Ectoine by a Transgenic, Nonhalophilic Bacterium Appl. Envir. Microbiol., May 15, 2007; 73(10): 3343 - 3347. [Abstract] [Full Text] [PDF]

				R. Garcia-Estepa, M. Argandona, M. Reina-Bueno, N. Capote, F. Iglesias-Guerra, J. J. Nieto, and C. Vargas The ectD Gene, Which Is Involved in the Synthesis of the Compatible Solute Hydroxyectoine, Is Essential for Thermoprotection of the Halophilic Bacterium Chromohalobacter salexigens J. Bacteriol., June 1, 2006; 188(11): 3774 - 3784. [Abstract] [Full Text] [PDF]

				D. Kapfhammer, E. Karatan, K. J. Pflughoeft, and P. I. Watnick Role for Glycine Betaine Transport in Vibrio cholerae Osmoadaptation and Biofilm Formation within Microbial Communities Appl. Envir. Microbiol., July 1, 2005; 71(7): 3840 - 3847. [Abstract] [Full Text] [PDF]

				M. Jebbar, L. Sohn-Bosser, E. Bremer, T. Bernard, and C. Blanco Ectoine-Induced Proteins in Sinorhizobium meliloti Include an Ectoine ABC-Type Transporter Involved in Osmoprotection and Ectoine Catabolism J. Bacteriol., February 15, 2005; 187(4): 1293 - 1304. [Abstract] [Full Text] [PDF]

				M. I. Calderon, C. Vargas, F. Rojo, F. Iglesias-Guerra, L. N. Csonka, A. Ventosa, and J. J. Nieto Complex regulation of the synthesis of the compatible solute ectoine in the halophilic bacterium Chromohalobacter salexigens DSM 3043T Microbiology, September 1, 2004; 150(9): 3051 - 3063. [Abstract] [Full Text] [PDF]

				J. Prabhu, F. Schauwecker, N. Grammel, U. Keller, and M. Bernhard Functional Expression of the Ectoine Hydroxylase Gene (thpD) from Streptomyces chrysomallus in Halomonas elongata Appl. Envir. Microbiol., May 1, 2004; 70(5): 3130 - 3132. [Abstract] [Full Text] [PDF]

				K. J. Pflughoeft, K. Kierek, and P. I. Watnick Role of Ectoine in Vibrio cholerae Osmoadaptation Appl. Envir. Microbiol., October 1, 2003; 69(10): 5919 - 5927. [Abstract] [Full Text] [PDF]

				M. Manzanera, A. Garcia de Castro, A. Tondervik, M. Rayner-Brandes, A. R. Strom, and A. Tunnacliffe Hydroxyectoine Is Superior to Trehalose for Anhydrobiotic Engineering of Pseudomonas putida KT2440 Appl. Envir. Microbiol., September 1, 2002; 68(9): 4328 - 4333. [Abstract] [Full Text] [PDF]

				K. Grammann, A. Volke, and H. J. Kunte New Type of Osmoregulated Solute Transporter Identified in Halophilic Members of the Bacteria Domain: TRAP Transporter TeaABC Mediates Uptake of Ectoine and Hydroxyectoine in Halomonas elongata DSM 2581T J. Bacteriol., June 1, 2002; 184(11): 3078 - 3085. [Abstract] [Full Text] [PDF]

				A. U. Kuhlmann and E. Bremer Osmotically Regulated Synthesis of the Compatible Solute Ectoine in Bacillus pasteurii and Related Bacillus spp. Appl. Envir. Microbiol., February 1, 2002; 68(2): 772 - 783. [Abstract] [Full Text] [PDF]

				H. Nakayama, K. Yoshida, H. Ono, Y. Murooka, and A. Shinmyo Ectoine, the Compatible Solute of Halomonas elongata, Confers Hyperosmotic Tolerance in Cultured Tobacco Cells Plant Physiology, April 1, 2000; 122(4): 1239 - 1248. [Abstract] [Full Text]

				L. O. Martins, N. Empadinhas, J. D. Marugg, C. Miguel, C. Ferreira, M. S. da Costa, and H. Santos Biosynthesis of Mannosylglycerate in the Thermophilic Bacterium Rhodothermus marinus. BIOCHEMICAL AND GENETIC CHARACTERIZATION OF A MANNOSYLGLYCERATE SYNTHASE J. Biol. Chem., December 10, 1999; 274(50): 35407 - 35414. [Abstract] [Full Text] [PDF]

				A. Oren Bioenergetic Aspects of Halophilism Microbiol. Mol. Biol. Rev., June 1, 1999; 63(2): 334 - 348. [Abstract] [Full Text] [PDF]

				G. Malin, R. Iakobashvili, and A. Lapidot Effect of Tetrahydropyrimidine Derivatives on Protein-Nucleic Acids Interaction. TYPE II RESTRICTION ENDONUCLEASES AS A MODEL SYSTEM J. Biol. Chem., March 12, 1999; 274(11): 6920 - 6929. [Abstract] [Full Text] [PDF]

				A. Ventosa, J. J. Nieto, and A. Oren Biology of Moderately Halophilic Aerobic Bacteria Microbiol. Mol. Biol. Rev., June 1, 1998; 62(2): 504 - 544. [Abstract] [Full Text] [PDF]