Novel description of mcl-PHA biosynthesis by Pseudomonas chlororaphis from animal-derived waste.

A novel description of mcl-PHA biosynthesis by Ps. chlororaphis from tallow-based biodiesel as an inexpensive carbon feed stock is presented. Fermentation protocols, kinetic analysis, an efficient product recovery strategy, and product characterization are included. Maximum specific growth rates (μmax.) of 0.08 h(-1), 0.10 h(-1) and 0.13 h(-1), respectively, were achieved in three different fermentation set-ups. Volumetric productivity for mcl-PHA amounted to 0.071 g/L h, 0.094 g/L h and 0.138 g/L h, final intracellular PHA contents calculated from the sum of active biomass and PHA from 22.1 to 29.4 wt.-%, respectively. GC-FID analysis showed that the obtained biopolyester predominantly consists of 3-hydroxyoctanoate and 3-hydroxydecanoate, and, to a minor extent, 3-hydroxydodecanoate, 3-hydroxynonanoate, 3-hydroxyhexanoate, and 3-hydroxyheptanoate monomers. The overall distribution of the monomers remained similar, regardless to working volumes, biodiesel concentrations and pre-treatment of the inoculum.

[1]  T. Foglia,et al.  Biosynthesis of Medium-chain-length Poly(hydroxyalkanoates) from Soy Molasses , 2006, Biotechnology Letters.

[2]  J. Ackermann,et al.  Synthesis of Poly(3‐hydroxybutyrate‐co‐4‐hydrobutyrate) with a Target Mole Fraction of 4‐Hydroxybutyric Acid Units by Two‐Stage Continuous Cultivation of Delftia acidovorans P4a , 2005 .

[3]  C. C. Lin,et al.  PHBV production by Ralstonia eutropha in a continuous stirred tank reactor , 2005 .

[4]  Martin Koller,et al.  Production of polyhydroxyalkanoates from agricultural waste and surplus materials. , 2005, Biomacromolecules.

[5]  R. Lenz,et al.  The microbial production of poly(hydroxyalkanoates) from tallow , 1996, Applied Microbiology and Biotechnology.

[6]  R. Rivas,et al.  Reclassification of Pseudomonas aurantiaca as a synonym of Pseudomonas chlororaphis and proposal of three subspecies, P. chlororaphis subsp. chlororaphis subsp. nov., P. chlororaphis subsp. aureofaciens subsp. nov., comb. nov. and P. chlororaphis subsp. aurantiaca subsp. nov., comb. nov. , 2007, International journal of systematic and evolutionary microbiology.

[7]  E. Chiellini,et al.  Continuous production of poly([R]-3-hydroxybutyrate) by Cupriavidus necator in a multistage bioreactor cascade , 2011, Applied Microbiology and Biotechnology.

[8]  S. Panke,et al.  Analysis of Pseudomonas putida alkane-degradation gene clusters and flanking insertion sequences: evolution and regulation of the alk genes. , 2001, Microbiology.

[9]  M. Zinn Biosynthesis of Medium-Chain-Length Poly[( R )-3-hydroxyalkanoates] , 2010 .

[10]  R. M. Lafferty,et al.  A rapid gas chromatographic method for the determination of poly-β-hydroxybutyric acid in microbial biomass , 1978, European journal of applied microbiology and biotechnology.

[11]  T. Egli,et al.  Accumulation of poly[(R)-3-hydroxyalkanoates] in Pseudomonas oleovorans during growth in batch and chemostat culture with different carbon sources. , 2001, Biotechnology and bioengineering.

[12]  Melba Lee,et al.  A Study of the Nature, Growth and Control of Bacteria in Cutting Compounds , 1941, Journal of bacteriology.

[13]  Sang Yup Lee,et al.  Plastic bacteria? Progress and prospects for polyhydroxyalkanoate production in bacteria , 1996 .

[14]  Anna Salerno,et al.  Modern Biotechnological Polymer Synthesis: A Review , 2010 .

[15]  B. Witholt,et al.  Two-stage continuous process development for the production of medium-chain-length poly(3-hydroxyalkanoates). , 2001, Biotechnology and bioengineering.

[16]  Guo-Qiang Chen Industrial Production of PHA , 2010 .

[17]  Tajalli Keshavarz,et al.  Polyhydroxyalkanoates: bioplastics with a green agenda. , 2010, Current opinion in microbiology.

[18]  Guo-Qiang Chen,et al.  Plastics Completely Synthesized by Bacteria: Polyhydroxyalkanoates , 2010 .

[19]  Martin Koller,et al.  Process optimization for efficient biomediated PHA production from animal-based waste streams , 2012, Clean Technologies and Environmental Policy.

[20]  S H Lee,et al.  Production of medium-chain-length polyhydroxyalkanoates by high-cell-density cultivation of Pseudomonas putida under phosphorus limitation. , 2000, Biotechnology and bioengineering.

[21]  T. Foglia,et al.  Glucose/Lipid Mixed Substrates as a Means of Controlling the Properties of Medium Chain Length Poly(hydroxyalkanoates)† , 2001 .

[22]  G. Huisman,et al.  Reduction of Cell Lysate Viscosity during Processing of Poly(3-Hydroxyalkanoates) by Chromosomal Integration of the Staphylococcal Nuclease Gene in Pseudomonas putida , 1999, Applied and Environmental Microbiology.

[23]  C. Guoqiang,et al.  Synthesis of copolyesters consisting of medium-chain-length β-hydroxyalkanoates by Pseudomonas stutzeri 1317 , 2001 .

[24]  Young Baek Kim,et al.  Production of unusual bacterial polyesters by Pseudomonas oleovorans through cometabolism , 1992 .

[25]  B. Kim Production of medium chain length polyhydroxyalkanoates by fed-batch culture of Pseudomonas oleovorans , 2004, Biotechnology Letters.

[26]  Jian Chen,et al.  Kinetic studies on poly-3-hydroxybutyrate formation by Ralstonia eutropha in a two-stage continuous culture system , 2001 .

[27]  R. Bartha,et al.  Pattern of phenazine pigment production by a strain of Pseudomonas aeruginosa , 1978, Journal of bacteriology.

[28]  O. White,et al.  Complete genome sequence and comparative analysis of the metabolically versatile Pseudomonas putida KT2440. , 2002, Environmental microbiology.

[29]  V. Langlois,et al.  Bacterial poly-3-hydroxyalkenoates with epoxy groups in the side chains , 1997 .

[30]  A. Steinbüchel,et al.  Formation of polyesters consisting of medium-chain-length 3-hydroxyalkanoic acids from gluconate by Pseudomonas aeruginosa and other fluorescent pseudomonads , 1990, Applied and environmental microbiology.

[31]  G. Du,et al.  Continuous production of poly-3-hydroxybutyrate by Ralstonia eutropha in a two-stage culture system. , 2001, Journal of biotechnology.

[32]  M. Guay,et al.  Fed-batch production of unsaturated medium-chain-length polyhydroxyalkanoates with controlled composition by Pseudomonas putida KT2440 , 2009, Applied Microbiology and Biotechnology.

[33]  A. Plückthun,et al.  Engineering novel binding proteins from nonimmunoglobulin domains , 2005, Nature Biotechnology.

[34]  M. Reis,et al.  Polyhydroxyalkanoate (PHA) production by a mixed microbial culture using sugar molasses: effect of the influent substrate concentration on culture selection. , 2010, Water research.

[35]  M. Koller,et al.  Microbial PHA Production from Waste Raw Materials , 2010 .

[36]  T. Egli,et al.  Occurrence, synthesis and medical application of bacterial polyhydroxyalkanoate. , 2001, Advanced drug delivery reviews.

[37]  Anne Pohlmann,et al.  Genome sequence of the bioplastic-producing “Knallgas” bacterium Ralstonia eutropha H16 , 2006, Nature Biotechnology.

[38]  M. Schmid,et al.  Tailored Synthesis of Poly([R]‐3‐hydroxybutyrate‐co‐3‐hydroxyvalerate) (PHB/HV) in Ralstonia eutropha DSM 428 , 2003 .

[39]  Y. Rhee,et al.  Overexpression of the (R)-Specific Enoyl-CoA Hydratase Gene from Pseudomonas chlororaphis HS21 in Pseudomonas Strains for the Biosynthesis of Polyhydroxyalkanoates of Altered Monomer Composition , 2012, Bioscience, biotechnology, and biochemistry.

[40]  K. Sudesh,et al.  Biosynthesis and characterization of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) and poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) copolymers using jatropha oil as the main carbon source , 2011 .

[41]  G. Braunegg,et al.  Polyhydroxyalkanoates, biopolyesters from renewable resources: physiological and engineering aspects. , 1998, Journal of biotechnology.

[42]  C. Rossell,et al.  Integrated production of biodegradable plastic, sugar and ethanol , 2001, Applied Microbiology and Biotechnology.

[43]  George Guo-Qiang Chen Plastics from Bacteria , 2010 .

[44]  J. Nielsen,et al.  Phenazine natural products: biosynthesis, synthetic analogues, and biological activity. , 2004, Chemical reviews.

[45]  T. Foglia,et al.  Conversion of agricultural feedstock and coproducts into poly(hydroxyalkanoates) , 2006, Applied Microbiology and Biotechnology.

[46]  B. Witholt,et al.  Tailor‐made olefinic medium‐chain‐length poly[(R)‐3‐hydroxyalkanoates] by Pseudomonas putida GPo1: Batch versus chemostat production , 2006, Biotechnology and bioengineering.

[47]  M. Choi,et al.  Accumulation of Polyhydroxyalkanoic Acid Containing Large Amounts of Unsaturated Monomers in Pseudomonas fluorescens BM07 Utilizing Saccharides and Its Inhibition by 2-Bromooctanoic Acid , 2001, Applied and Environmental Microbiology.

[48]  B. Witholt,et al.  Poly(3-hydroxyalkanoate) polymerase synthesis and in vitro activity in recombinant Escherichia coli and Pseudomonas putida , 2005, Applied Microbiology and Biotechnology.

[49]  M. Koller,et al.  POLYHYDROXYALKANOATES: BIODEGRADABLE POLYMERIC MATERIALS FROM RENEWABLE RESOURCES , 2012 .

[50]  S. Panke,et al.  Efficient recovery of low endotoxin medium-chain-length poly([R]-3-hydroxyalkanoate) from bacterial biomass. , 2007, Journal of microbiological methods.

[51]  Anna Salerno,et al.  Biodegradable Latexes from Animal-Derived Waste: Biosynthesis and Characterization of mcl-PHA accumulated by Ps. citronellolis , 2013 .