This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Liebl, W Articles by Schleifer, K H Search for Related Content PubMed PubMed Citation Articles by Liebl, W Articles by Schleifer, K H

research-article

Expression, secretion, and processing of staphylococcal nuclease by Corynebacterium glutamicum.

Lehrstuhl für Mikrobiologie, Technische Universität München, Germany.

ABSTRACT

The gene for staphylococcal nuclease (SNase), an extracellular enzyme of Staphylococcus aureus, was introduced into Corynebacterium glutamicum. The heterologous gene was expressed in this host organism, and SNase was efficiently exported to the culture medium. Amino-terminal sequencing of SNase secreted by C. glutamicum revealed that the signal peptide was apparently cleaved off at precisely the same position as in the original host, S. aureus. As with S. aureus, a second smaller form of SNase (A form), whose appearance is presumably the result of a secondary processing step, was found in the culture medium of the recombinant C. glutamicum strain. The A form was one residue shorter than the mature nuclease A produced by S. aureus. Variation of the sodium chloride concentration in the growth medium had a marked influence on the location and the processing of SNase by C. glutamicum. In a complex growth medium containing 4% sodium chloride, SNase was exclusively located in the supernatant, but a significant amount of the enzyme remained cell associated if the strain was grown in a low-salt medium. Also, high salt concentrations seemed to inhibit processing of the high-molecular-weight form of SNase (B form) to the smaller A form. Similarities and differences in the export and modes of processing of SNase by three different, nonrelated gram-positive host organisms are discussed. Finally, a versatile Escherichia coli-C. glutamicum tac-lacIq expression shuttle vector was constructed. With this vector, it was possible to achieve isopropyl-beta-D-galactopyranoside (IPTG)-inducible overexpression and secretion of SNase in C. glutamicum, whereby the expression level was dependent on the concentration of the inducer.

This article has been cited by other articles:

• Jakob, K., Satorhelyi, P., Lange, C., Wendisch, V. F., Silakowski, B., Scherer, S., Neuhaus, K. (2007). Gene Expression Analysis of Corynebacterium glutamicum Subjected to Long-Term Lactic Acid Adaptation. J. Bacteriol. 189: 5582-5590 [Abstract] [Full Text]  
• Brinster, S., Furlan, S., Serror, P. (2007). C-Terminal WxL Domain Mediates Cell Wall Binding in Enterococcus faecalis and Other Gram-Positive Bacteria. J. Bacteriol. 189: 1244-1253 [Abstract] [Full Text]  
• Mayr, U. B., Haller, C., Haidinger, W., Atrasheuskaya, A., Bukin, E., Lubitz, W., Ignatyev, G. (2005). Bacterial Ghosts as an Oral Vaccine: a Single Dose of Escherichia coli O157:H7 Bacterial Ghosts Protects Mice against Lethal Challenge. Infect. Immun. 73: 4810-4817 [Abstract] [Full Text]  
• MacConaill, L. E., Fitzgerald, G. F., van Sinderen, D. (2003). Investigation of Protein Export in Bifidobacterium breve UCC2003. Appl. Environ. Microbiol. 69: 6994-7001 [Abstract] [Full Text]  
• Haidinger, W., Mayr, U. B., Szostak, M. P., Resch, S., Lubitz, W. (2003). Escherichia coli Ghost Production by Expression of Lysis Gene E and Staphylococcal Nuclease. Appl. Environ. Microbiol. 69: 6106-6113 [Abstract] [Full Text]  
• Tzvetkov, M., Klopprogge, C., Zelder, O., Liebl, W. (2003). Genetic dissection of trehalose biosynthesis in Corynebacterium glutamicum: inactivation of trehalose production leads to impaired growth and an altered cell wall lipid composition. Microbiology 149: 1659-1673 [Abstract] [Full Text]  
• Kikuchi, Y., Date, M., Yokoyama, K.-i., Umezawa, Y., Matsui, H. (2003). Secretion of Active-Form Streptoverticillium mobaraense Transglutaminase by Corynebacterium glutamicum: Processing of the Pro-Transglutaminase by a Cosecreted Subtilisin-Like Protease from Streptomyces albogriseolus. Appl. Environ. Microbiol. 69: 358-366 [Abstract] [Full Text]  
• Recchi, C., Rauzier, J., Gicquel, B., Reyrat, J.-M. (2002). Signal-sequence-independent secretion of the staphylococcal nuclease in Mycobacterium smegmatis. Microbiology 148: 529-536 [Abstract] [Full Text]  
• Le Loir, Y., Nouaille, S., Commissaire, J., Bretigny, L., Gruss, A., Langella, P. (2001). Signal Peptide and Propeptide Optimization for Heterologous Protein Secretion in Lactococcus lactis. Appl. Environ. Microbiol. 67: 4119-4127 [Abstract] [Full Text]  
• Dieye, Y., Usai, S., Clier, F., Gruss, A., Piard, J.-C. (2001). Design of a Protein-Targeting System for Lactic Acid Bacteria. J. Bacteriol. 183: 4157-4166 [Abstract] [Full Text]  
• Boynton, Z. L., Koon, J. J., Brennan, E. M., Clouart, J. D., Horowitz, D. M., Gerngross, T. U., Huisman, G. W. (1999). Reduction of Cell Lysate Viscosity during Processing of Poly(3-Hydroxyalkanoates) by Chromosomal Integration of the Staphylococcal Nuclease Gene in Pseudomonas putida. Appl. Environ. Microbiol. 65: 1524-1529 [Abstract] [Full Text]  
• Le Loir, Y., Gruss, A., Ehrlich, S. D., Langella, P. (1998). A Nine-Residue Synthetic Propeptide Enhances Secretion Efficiency of Heterologous Proteins in Lactococcus lactis. J. Bacteriol. 180: 1895-1903 [Abstract] [Full Text]  
• Poquet, I., Ehrlich, S. D., Gruss, A. (1998). An Export-Specific Reporter Designed for Gram-Positive Bacteria: Application to Lactococcus lactis. J. Bacteriol. 180: 1904-1912 [Abstract] [Full Text]