Engineering the robustness of Clostridium acetobutylicum by introducing glutathione biosynthetic capability.

To improve the aero- and solvent tolerance of the solvent-producing Clostridium acetobutylicum, glutathione biosynthetic capability was introduced into C. acetobutylicum DSM1731 by cloning and over-expressing the gshAB genes from Escherichia coli. Strain DSM1731(pITAB) produces glutathione, and shows a significantly improved survival upon aeration and butanol challenge, as compared with the control. In addition, strain DSM1731(pITAB) exhibited an improved butanol tolerance and an increased butanol production capability, as compared with the recombinant strains with only gshA or gshB gene. These results illustrated that introducing glutathione biosynthetic pathway, which is redundant for the metabolism of C. acetobutylicum, can increase the robustness of the host to achieve a better solvent production.

[1]  L N Csonka,et al.  Physiological and genetic responses of bacteria to osmotic stress. , 1989, Microbiological reviews.

[2]  P. Goffin,et al.  Improved adaptation to heat, cold, and solvent tolerance in Lactobacillus plantarum , 2007, Applied Microbiology and Biotechnology.

[3]  H. Bahl,et al.  Desulfoferrodoxin of Clostridium acetobutylicum functions as a superoxide reductase , 2007, FEBS letters.

[4]  O. Griffith,et al.  Glutathione Synthesis in Streptococcus agalactiae , 2005, Journal of Biological Chemistry.

[5]  E. Papoutsakis,et al.  Metabolic engineering of the non-sporulating, non-solventogenic Clostridium acetobutylicum strain M5 to produce butanol without acetone demonstrate the robustness of the acid-formation pathways and the importance of the electron balance. , 2008, Metabolic engineering.

[6]  Rainer Kalscheuer,et al.  Microdiesel: Escherichia coli engineered for fuel production. , 2006, Microbiology.

[7]  Kevin M. Smith,et al.  Metabolic engineering of Escherichia coli for 1-butanol production. , 2008, Metabolic engineering.

[8]  H. Bahl,et al.  The Role of PerR in O2-Affected Gene Expression of Clostridium acetobutylicum , 2009, Journal of bacteriology.

[9]  R. P. Ross,et al.  Improved Stress Tolerance of GroESL-Overproducing Lactococcus lactis and Probiotic Lactobacillus paracasei NFBC 338 , 2004, Applied and Environmental Microbiology.

[10]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[11]  M. Kleerebezem,et al.  Introducing glutathione biosynthetic capability into Lactococcus lactis subsp. cremoris NZ9000 improves the oxidative-stress resistance of the host. , 2006, Metabolic engineering.

[12]  H. Bahl,et al.  Reductive dioxygen scavenging by flavo‐diiron proteins of Clostridium acetobutylicum , 2009, FEBS letters.

[13]  E. Papoutsakis,et al.  A comparative view of metabolite and substrate stress and tolerance in microbial bioprocessing: From biofuels and chemicals, to biocatalysis and bioremediation. , 2010, Metabolic engineering.

[14]  A. Schirmer,et al.  Microbial Biosynthesis of Alkanes , 2010, Science.

[15]  H. Bahl,et al.  Pathway for H2O2 and O2 detoxification in Clostridium acetobutylicum. , 2009, Microbiology.

[16]  E. Papoutsakis,et al.  A genomic-library based discovery of a novel, possibly synthetic, acid-tolerance mechanism in Clostridium acetobutylicum involving non-coding RNAs and ribosomal RNA processing. , 2010, Metabolic engineering.

[17]  Kouji Takeda,et al.  Adaptive Responses to Oxygen Stress in Obligatory Anaerobes Clostridium acetobutylicum and Clostridium aminovalericum , 2005, Applied and Environmental Microbiology.

[18]  W. Goebel,et al.  A Multidomain Fusion Protein in Listeria monocytogenes Catalyzes the Two Primary Activities for Glutathione Biosynthesis , 2005, Journal of bacteriology.

[19]  P. Dürre,et al.  Changes in protein synthesis and identification of proteins specifically induced during solventogenesis in Clostridium acetobutylicum , 2002, Electrophoresis.

[20]  Eva R. Kashket,et al.  Intracellular Conditions Required for Initiation of Solvent Production by Clostridium acetobutylicum , 1986, Applied and environmental microbiology.

[21]  Nathan D. Price,et al.  Achievements and perspectives to overcome the poor solvent resistance in acetone and butanol-producing microorganisms , 2010, Applied Microbiology and Biotechnology.

[22]  E. Papoutsakis,et al.  Expression of Cloned Homologous Fermentative Genes in Clostridium Acetobutylicum ATCC 824 , 1992, Bio/Technology.

[23]  Host-plasmid interactions in recombinant strains of Clostridium acetobutylicum ATCC 824 , 1994 .

[24]  A. Zeng,et al.  Bulk chemicals from biotechnology: the case of 1,3-propanediol production and the new trends. , 2002, Advances in biochemical engineering/biotechnology.

[25]  H. Bahl,et al.  PerR acts as a switch for oxygen tolerance in the strict anaerobe Clostridium acetobutylicum , 2008, Molecular microbiology.

[26]  E. Papoutsakis,et al.  Dynamics of Genomic-Library Enrichment and Identification of Solvent Tolerance Genes for Clostridium acetobutylicum , 2007, Applied and Environmental Microbiology.

[27]  E. Papoutsakis,et al.  Metabolite stress and tolerance in the production of biofuels and chemicals: Gene‐expression‐based systems analysis of butanol, butyrate, and acetate stresses in the anaerobe Clostridium acetobutylicum , 2010, Biotechnology and bioengineering.

[28]  J. Hugenholtz,et al.  Glutathione Protects Lactococcus lactis against Acid Stress , 2007, Applied and Environmental Microbiology.

[29]  H. Yamaguchi,et al.  Three-dimensional structure of the glutathione synthetase from Escherichia coli B at 2.0 A resolution. , 1994, Journal of molecular biology.

[30]  Eleftherios T. Papoutsakis,et al.  Transcriptional Program of Early Sporulation and Stationary-Phase Events in Clostridium acetobutylicum , 2005, Journal of bacteriology.

[31]  E. Papoutsakis Engineering solventogenic clostridia. , 2008, Current opinion in biotechnology.

[32]  Yanping Zhang,et al.  Proteomic Analyses To Reveal the Protective Role of Glutathione in Resistance of Lactococcus lactis to Osmotic Stress , 2010, Applied and Environmental Microbiology.

[33]  L. Huang,et al.  Transmembrane pH gradient and membrane potential in Clostridium acetobutylicum during growth under acetogenic and solventogenic conditions , 1985, Applied and environmental microbiology.

[34]  C. Tomas,et al.  Overexpression of groESL in Clostridium acetobutylicum Results in Increased Solvent Production and Tolerance, Prolonged Metabolism, and Changes in the Cell's Transcriptional Program , 2003, Applied and Environmental Microbiology.

[35]  A. Meister,et al.  On the active site thiol of gamma-glutamylcysteine synthetase: relationships to catalysis, inhibition, and regulation. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[36]  R. C. Fahey,et al.  Copyright © 1998, American Society for Microbiology Import and Metabolism of Glutathione by Streptococcus mutans , 1997 .

[37]  D. Molenaar,et al.  Glutathione Protects Lactococcus lactis against Oxidative Stress , 2003, Applied and Environmental Microbiology.

[38]  C. Cha,et al.  Synthesis of gamma-glutamylcysteine as a major low-molecular-weight thiol in lactic acid bacteria Leuconostoc spp. , 2008, Biochemical and biophysical research communications.

[39]  Yanping Zhang,et al.  Engineering Clostridium Strain to Accept Unmethylated DNA , 2010, PloS one.

[40]  W. B. Adams,et al.  Occurrence of glutathione in bacteria , 1978, Journal of bacteriology.

[41]  Shelly C. Lu Regulation of glutathione synthesis. , 2009, Molecular aspects of medicine.

[42]  A. Meister Glutathione metabolism and its selective modification. , 1988, The Journal of biological chemistry.

[43]  J. Strahler,et al.  Quantifying changes in the thiol redox proteome upon oxidative stress in vivo , 2008, Proceedings of the National Academy of Sciences.

[44]  G. Smirnova,et al.  The role of antioxidant enzymes in response of Escherichia coli to osmotic upshift. , 2000, FEMS microbiology letters.

[45]  Hiroaki Kitano,et al.  Biological robustness , 2008, Nature Reviews Genetics.

[46]  J. Liao,et al.  Non-fermentative pathways for synthesis of branched-chain higher alcohols as biofuels , 2008, Nature.

[47]  G. Georgiou,et al.  The many faces of glutathione in bacteria. , 2006, Antioxidants & redox signaling.

[48]  K. Prather,et al.  Engineering alternative butanol production platforms in heterologous bacteria. , 2009, Metabolic engineering.

[49]  Ming Liu,et al.  Inactivation of aldehyde dehydrogenase: a key factor for engineering 1,3-propanediol production by Klebsiella pneumoniae. , 2006, Metabolic engineering.

[50]  L. K. Bowles,et al.  Effects of butanol on Clostridium acetobutylicum , 1985, Applied and environmental microbiology.

[51]  G. Smirnova,et al.  Role of Glutathione in the Response of Escherichia coli to Osmotic Stress , 2001, Biochemistry (Moscow).

[52]  I. Schuppe-Koistinen,et al.  S-glutathionylation of glyceraldehyde-3-phosphate dehydrogenase: role of thiol oxidation and catalysis by glutaredoxin. , 2002, Methods in enzymology.

[53]  E. Papoutsakis,et al.  Thiolase from Clostridium acetobutylicum ATCC 824 and Its Role in the Synthesis of Acids and Solvents , 1988, Applied and environmental microbiology.

[54]  Zugen Chen,et al.  Proteome reference map and comparative proteomic analysis between a wild type Clostridium acetobutylicum DSM 1731 and its mutant with enhanced butanol tolerance and butanol yield. , 2010, Journal of proteome research.

[55]  S. Kawasaki,et al.  Identification of O2‐induced peptides in an obligatory anaerobe, Clostridium acetobutylicum , 2004, FEBS letters.

[56]  Jay D. Keasling,et al.  Functional Genomic Study of Exogenous n-Butanol Stress in Escherichia coli , 2010, Applied and Environmental Microbiology.

[57]  D. T. Jones,et al.  Acetone-butanol fermentation revisited. , 1986, Microbiological reviews.

[58]  Yan Zhu,et al.  The Importance of Engineering Physiological Functionality into Microbes Opinion , 2022 .