Adaptation of Corynebacterium glutamicum to salt‐stress conditions

Corynebacterium glutamicum is one of the biotechnologically most important microorganisms because of its ability to enrich amino acids extracellularly. Hence, C. glutamicum requires effective adaptation strategies against both hypo‐ and hyperosmotic stress. We give a comprehensive and coherent outline about the quantitative dynamics of C. glutamicum during adaptation to hyperosmotic stress at the transcript and protein levels. The osmolyte carrier ProP, playing a pivotal role in hyperosmotic stress defence, exhibits the strongest up‐regulation of all proteins. A conspicuously regulated group comprises proteins involved in lipid biosynthesis of the cell envelope. This is in accordance with our observation of a more viscous and stickier cell envelope, which is supported by the findings of an altered lipid composition. Together with our results, showing that several transporters were down‐regulated, this membrane adaptation appears to be one of C. glutamicum's major protection strategies against hyperosmotic stress. In addition, we demonstrate that no oxidative stress and no iron limitation occur during salt stress contrary to former postulations. Ultimately, it is remarkable that various proteins with divergent mRNA‐protein dynamics and regulation have been observed. This leads to the assumption that there are still unknown mechanisms in between the bacterial transcription, translation and post‐translation and that these are waiting to be unravelled.

[1]  Wayne F. Patton,et al.  Ultrasensitive fluorescence protein detection in isoelectric focusing gels using a ruthenium metal chelate stain , 2000, Electrophoresis.

[2]  Hyungjoon Kim,et al.  Functional analysis of sigH expression in Corynebacterium glutamicum. , 2005, Biochemical and biophysical research communications.

[3]  Wei-Shou Hu,et al.  Uncovering Genes with Divergent mRNA-Protein Dynamics in Streptomyces coelicolor , 2008, PloS one.

[4]  Christer S. Ejsing,et al.  Osmolality, Temperature, and Membrane Lipid Composition Modulate the Activity of Betaine Transporter BetP in Corynebacterium glutamicum , 2007, Journal of bacteriology.

[5]  I. Shiio,et al.  Methionine biosynthesis in Brevibacterium flavum: properties and essential role of O-acetylhomoserine sulfhydrylase. , 1982, Journal of biochemistry.

[6]  Analysis of Corynebacterium glutamicum methionine biosynthetic pathway: isolation and analysis of metB encoding cystathionine gamma-synthase. , 1999, Molecules and cells.

[7]  R. Krämer,et al.  Characterization of compatible solute transporter multiplicity in Corynebacterium glutamicum , 2007, Applied Microbiology and Biotechnology.

[8]  Y. Kim,et al.  Isolation and analysis of metA, a methionine biosynthetic gene encoding homoserine acetyltransferase in corynebacterium glutamicum. , 1998, Molecules and cells.

[9]  J. Kalinowski,et al.  Development of a Corynebacterium glutamicum DNA microarray and validation by genome-wide expression profiling during growth with propionate as carbon source. , 2003, Journal of biotechnology.

[10]  D. Wolters,et al.  Bacterial membrane proteomics , 2008, Proteomics.

[11]  Alexander Sczyrba,et al.  EMMA: a platform for consistent storage and efficient analysis of microarray data. , 2003, Journal of biotechnology.

[12]  M. Taylor,et al.  Molecular analyses of mycobacteria other than the M. tuberculosis complex isolated from Northern Ireland cattle. , 2005, Veterinary microbiology.

[13]  M. Salton The Bacterial Membrane , 1971 .

[14]  S. Morbach,et al.  Three pathways for trehalose metabolism in Corynebacterium glutamicum ATCC13032 and their significance in response to osmotic stress , 2003, Molecular microbiology.

[15]  M. Bott,et al.  Deletion of the genes encoding the MtrA–MtrB two‐component system of Corynebacterium glutamicum has a strong influence on cell morphology, antibiotics susceptibility and expression of genes involved in osmoprotection , 2004, Molecular microbiology.

[16]  M. Wada,et al.  Purification and Characterization of O-Acetylserine Sulfhydrylase of Corynebacterium glutamicum , 2004, Bioscience, biotechnology, and biochemistry.

[17]  G. Besra,et al.  Acyl-CoA Carboxylases (accD2 and accD3), Together with a Unique Polyketide Synthase (Cg-pks), Are Key to Mycolic Acid Biosynthesis in Corynebacterianeae Such as Corynebacterium glutamicum and Mycobacterium tuberculosis* , 2004, Journal of Biological Chemistry.

[18]  A. Pühler,et al.  Role of the ssu and seu Genes of Corynebacterium glutamicum ATCC 13032 in Utilization of Sulfonates and Sulfonate Esters as Sulfur Sources , 2005, Applied and Environmental Microbiology.

[19]  G. Besra,et al.  The Two Carboxylases of Corynebacterium glutamicum Essential for Fatty Acid and Mycolic Acid Synthesis , 2007, Journal of bacteriology.

[20]  Reinhard Krämer,et al.  Impact of osmotic stress on volume regulation, cytoplasmic solute composition and lysine production in Corynebacterium glutamicum MH20-22B. , 2003, Journal of biotechnology.

[21]  K. Jung,et al.  Time-Dependent Proteome Alterations under Osmotic Stress during Aerobic and Anaerobic Growth in Escherichia coli , 2006, Journal of bacteriology.

[22]  A. Burkovski,et al.  GltS, the sodium-coupled L-glutamate uptake system of Corynebacterium glutamicum: identification of the corresponding gene and impact on L-glutamate production , 2003, Applied Microbiology and Biotechnology.

[23]  D. Richards,et al.  Development and applications of in-gel CNBr/tryptic digestion combined with mass spectrometry for the analysis of membrane proteins. , 2003, Journal of proteome research.

[24]  M. Chami,et al.  Mycomembrane and S-layer: two important structures of Corynebacterium glutamicum cell envelope with promising biotechnology applications. , 2003, Journal of biotechnology.

[25]  J. Kalinowski,et al.  Functional genomics and expression analysis of the Corynebacterium glutamicum fpr2-cysIXHDNYZ gene cluster involved in assimilatory sulphate reduction , 2005, BMC Genomics.

[26]  L. Eggeling,et al.  Handbook of Corynebacterium glutamicum , 2005 .

[27]  H. Kase,et al.  Production of L-Threonine by Analog-resistant Mutants , 1972 .

[28]  J. Kalinowski,et al.  Classification of hyper-variable Corynebacterium glutamicum surface-layer proteins by sequence analyses and atomic force microscopy. , 2004, Journal of biotechnology.

[29]  D. Wolters,et al.  The two‐phase partitioning system – a powerful technique to purify integral membrane proteins of Corynebacterium glutamicum for quantitative shotgun analysis , 2009, Proteomics.

[30]  J. Kalinowski,et al.  Identification and functional analysis of six mycolyltransferase genes of Corynebacterium glutamicum ATCC 13032: the genes cop1, cmt1, and cmt2 can replace each other in the synthesis of trehalose dicorynomycolate, a component of the mycolic acid layer of the cell envelope , 2003, Archives of Microbiology.

[31]  B. Hwang,et al.  Corynebacterium glutamicum Utilizes both Transsulfuration and Direct Sulfhydrylation Pathways for Methionine Biosynthesis , 2002, Journal of bacteriology.

[32]  J. Kalinowski,et al.  The McbR repressor modulated by the effector substance S‐adenosylhomocysteine controls directly the transcription of a regulon involved in sulphur metabolism of Corynebacterium glutamicum ATCC 13032 , 2005, Molecular microbiology.

[33]  A. Krogh,et al.  Predicting transmembrane protein topology with a hidden Markov model: application to complete genomes. , 2001, Journal of molecular biology.

[34]  C. Raetz,et al.  Origin of Lipid A Species Modified with 4-Amino-4-deoxy-l-arabinose in Polymyxin-resistant Mutants of Escherichia coli , 2003, Journal of Biological Chemistry.

[35]  A. Pühler,et al.  Genome-wide analysis of the L-methionine biosynthetic pathway in Corynebacterium glutamicum by targeted gene deletion and homologous complementation. , 2003, Journal of biotechnology.

[36]  Michael Hecker,et al.  From complementarity to comprehensiveness – targeting the membrane proteome of growing Bacillus subtilis by divergent approaches , 2008, Proteomics.

[37]  M. Bott,et al.  clpC and clpP1P2 gene expression in Corynebacterium glutamicum is controlled by a regulatory network involving the transcriptional regulators ClgR and HspR as well as the ECF sigma factor σH , 2004, Molecular microbiology.

[38]  Rapid cloning of metK encoding methionine adenosyltransferase from Corynebacterium glutamicum by screening a genomic library on a high density colony-array. , 2000, FEMS microbiology letters.

[39]  Jörg Bernhardt,et al.  Salt stress adaptation of Bacillus subtilis: A physiological proteomics approach , 2006, Proteomics.

[40]  S. Amar,et al.  Proteomic mapping of stimulus-specific signaling pathways involved in THP-1 cells exposed to Porphyromonas gingivalis or its purified components. , 2007, Journal of proteome research.

[41]  B. Eikmanns,et al.  Identification and Characterization of a Bacterial Transport System for the Uptake of Pyruvate, Propionate, and Acetate in Corynebacterium glutamicum , 2008, Journal of bacteriology.

[42]  M. Daffé,et al.  Characterization of the in vivo acceptors of the mycoloyl residues transferred by the corynebacterial PS1 and the related mycobacterial antigens 85 , 2000, Molecular microbiology.

[43]  J. Yates,et al.  A correlation algorithm for the automated quantitative analysis of shotgun proteomics data. , 2003, Analytical chemistry.

[44]  David L Tabb,et al.  ProRata: A quantitative proteomics program for accurate protein abundance ratio estimation with confidence interval evaluation. , 2006, Analytical chemistry.

[45]  O. Sorgenfrei,et al.  Genome-wide transcription profiling of Corynebacterium glutamicum after heat shock and during growth on acetate and glucose. , 2002, Journal of biotechnology.

[46]  U. Völker,et al.  High-Salinity-Induced Iron Limitation in Bacillus subtilis , 2002, Journal of bacteriology.

[47]  H. Kase,et al.  The regulation of L-methionine synthesis and the properties of cystathionine .ALPHA.-synthase and .BETA.-cystathionase in Corynebacterium glutamicum. , 1974 .

[48]  Ivan Mijakovic,et al.  MATERIALS AND METHODS , 1981, Green Corrosion Inhibitors: Reviews and Applications.

[49]  A. Goesmann,et al.  The complete Corynebacterium glutamicum ATCC 13032 genome sequence and its impact on the production of L-aspartate-derived amino acids and vitamins. , 2003, Journal of biotechnology.

[50]  E. Kimura,et al.  A dtsR gene-disrupted mutant of Brevibacterium lactofermentum requires fatty acids for growth and efficiently produces L-glutamate in the presence of an excess of biotin. , 1997, Biochemical and biophysical research communications.

[51]  F. Fischer,et al.  Proteome Science BioMed Central , 2004 .

[52]  J. Kalinowski,et al.  Functional genomics of pH homeostasis in Corynebacterium glutamicum revealed novel links between pH response, oxidative stress, iron homeostasis and methionine synthesis , 2009, BMC Genomics.

[53]  Vassily Hatzimanikatis,et al.  Insights into the relation between mRNA and protein expression patterns: I. theoretical considerations , 2003, Biotechnology and bioengineering.

[54]  J. Kalinowski,et al.  The DtxR protein acting as dual transcriptional regulator directs a global regulatory network involved in iron metabolism of Corynebacterium glutamicum , 2006, BMC Genomics.