This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Hai, T.
Right arrow Articles by Steinbüchel, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Hai, T.
Right arrow Articles by Steinbüchel, A.
Agricola
Right arrow Articles by Hai, T.
Right arrow Articles by Steinbüchel, A.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, January 2002, p. 93-101, Vol. 68, No. 1
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.1.93-101.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Molecular Characterization of a Thermostable Cyanophycin Synthetase from the Thermophilic Cyanobacterium Synechococcus sp. Strain MA19 and In Vitro Synthesis of Cyanophycin and Related Polyamides

Tran Hai, Fred Bernd Oppermann-Sanio, and Alexander Steinbüchel*

Institut für Mikrobiologie, Westfälische Wilhelms-Universität, D-48149 Münster, Germany

Received 29 June 2001/ Accepted 26 September 2001

The thermophilic cyanobacterium Synechococcus sp. strain MA19 contained the structural genes for cyanophycin synthetase (cphA) and cyanophycinase (cphB), which were identified, cloned, and sequenced in this study. The translation products of cphA and cphB exhibited high levels of similarity to corresponding proteins of other cyanobacteria, such as Anabaena variabilis and Synechocystis sp. Recombinant cells of Escherichia coli harboring cphA colinear with lacPO accumulated cyanophycin that accounted for up to 25% (wt/wt) of the dry cell matter in the presence of isopropyl-ß-D-thiogalactopyranoside (IPTG). The cyanophycin synthetase was enriched 123-fold to electrophoretic homogeneity from the soluble fraction of the recombinant cells by anion-exchange chromatography, affinity chromatography, and gel filtration chromatography. The purified cyanophycin synthetase maintained the parental thermophilic character and was active even after prolonged incubation at 50°C; in the presence of ectoine the enzyme retained 90% of its activity even after 2 h of incubation. The in vitro activity of the enzyme depended on ATP, primers, and both substrates, L-arginine and L-aspartic acid. In addition to native cyanophycin, the purified enzyme accepted a modified cyanophycin containing less arginine, {alpha}-arginyl aspartic acid dipeptide, and poly-{alpha},ß-DL-aspartic acid as primers and also incorporated ß-hydroxyaspartic acid instead of L-aspartic acid or L-canavanine instead of L-arginine at a significant rate. The lack of specificity of this thermostable enzyme with respect to primers and substrates, the thermal stability of the enzyme, and the finding that the enzyme is suitable for in vitro production of cyanophycin make it an interesting candidate for biotechnological processes.

* Corresponding author. Mailing address: Institut für Mikrobiologie der Westfälischen Wilhelms-Universität, Corrensstrasse 3, D-48149 Münster, Germany. Phone: 49 251 8339821. Fax: 49 251 8338388. Email: steinbu{at}uni-muenster.de.

Applied and Environmental Microbiology, January 2002, p. 93-101, Vol. 68, No. 1
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.1.93-101.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Steinle, A., Bergander, K., Steinbuchel, A. (2009). Metabolic Engineering of Saccharomyces cerevisiae for Production of Novel Cyanophycins with an Extended Range of Constituent Amino Acids. Appl. Environ. Microbiol. 75: 3437-3446 [Abstract] [Full Text]  
  • Steinle, A., Oppermann-Sanio, F. B., Reichelt, R., Steinbuchel, A. (2008). Synthesis and Accumulation of Cyanophycin in Transgenic Strains of Saccharomyces cerevisiae. Appl. Environ. Microbiol. 74: 3410-3418 [Abstract] [Full Text]  
  • Hai, T., Frey, K. M., Steinbuchel, A. (2006). Engineered Cyanophycin Synthetase (CphA) from Nostoc ellipsosporum Confers Enhanced CphA Activity and Cyanophycin Accumulation to Escherichia coli. Appl. Environ. Microbiol. 72: 7652-7660 [Abstract] [Full Text]  
  • Obst, M., Krug, A., Luftmann, H., Steinbuchel, A. (2005). Degradation of Cyanophycin by Sedimentibacter hongkongensis Strain KI and Citrobacter amalonaticus Strain G Isolated from an Anaerobic Bacterial Consortium. Appl. Environ. Microbiol. 71: 3642-3652 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»