Physiology, regulation, and limits of the synthesis of poly(3HB).

The properties of poly(3-hydroxybutyrate) combined with the fact that it can be produced easily by numerous prokaryotes from renewable resources and even from potentially toxic waste products using well-known fermentation processes have generated keen interest in this biopolyester as a substitute for chemo-synthetic petroleum-derived polymers in many applications. However, the high price of poly(3HB) compared with the conventional synthetic materials currently in use has restricted its availability in a wide range of applications. If the economic viability of poly(3HB) production and its competitiveness are to be improved, more must be found out about the phenotypic optimization and the upper limits of bacterial systems as the factory of poly(3HB). In this chapter, two aspects of poly(3HB) are reviewed--poly(3HB) formation as a physiological response to external limitations and overcoming internal bottlenecks, and poly(3HB) as a commercially attractive polyester. From a physiological viewpoint, the ability to synthesize and degrade poly(3HB) is considered an investment in the future and provides organisms with a selective advantage. Poly(3HB) is presented as a strategic survival polymer, and it is shown that growth-associated synthesis is not as rare as reported. The influence of the efficiency and velocity of cell multiplication and product formation, of poly(3HB) content and of productivity on the overall yield, and finally on the economics of the whole process are discussed and evaluated from the technological or consumer's point of view. The specific production rate and poly(3HB) content appear to be more important than the yield coefficients.

[1]  M. Duchars,et al.  The Influence of C:N Ratio in the Growth Medium on the Cellular Composition and Regulation of Enzyme Activity in Hyphomicrobium X , 1989 .

[2]  S. Lee,et al.  Poly(3-Hydroxybutyrate) Production with High Productivity and High Polymer Content by a Fed-Batch Culture of Alcaligenes latus under Nitrogen Limitation , 1997, Applied and environmental microbiology.

[3]  D. Jendrossek Microbial degradation of polyesters: a review on extracellular poly(hydroxyalkanoic acid) depolymerases , 1998 .

[4]  Gjalt W. Huisman,et al.  Metabolic Engineering of Poly(3-Hydroxyalkanoates): From DNA to Plastic , 1999, Microbiology and Molecular Biology Reviews.

[5]  C. Föllner,et al.  Isolation and purification of granule-associated proteins relevant for poly(3-hydroxybutyric acid) biosynthesis from methylotrophic bacteria relying on the serine pathway , 1995 .

[6]  K. Sattler,et al.  Untersuchungen zur wachstumsassoziierten akkumulation von Poly-β-hydroxybuttersäure bei Methylobacterium rhodesianum Z , 1991 .

[7]  W. Babel,et al.  Mikrobielle Thermpolaste : Biosyntheses, Eigenschaften und Anwendung , 1990 .

[8]  D. E. Atkinson,et al.  Regulation of Enzyme Activity , 1966 .

[9]  D. Seebach,et al.  Synthese monodisperser linearer und cyclischer Oligomere der (R)‐3‐Hydroxybuttersäure mit bis zu 128 Einheiten , 1996 .

[10]  R. Gross,et al.  Plastics from bacteria and for bacteria: poly(beta-hydroxyalkanoates) as natural, biocompatible, and biodegradable polyesters. , 1990, Advances in biochemical engineering/biotechnology.

[11]  E. Dawes,et al.  Regulation of the tricarboxylic acid cycle and poly-beta-hydroxybutyrate metabolism in Azotobacter beijerinckii grown under nitrogen or oxygen limitation. , 1976, Journal of general microbiology.

[12]  P. Weitzman Unity and diversity in some bacterial citric acid-cycle enzymes. , 1981, Advances in microbial physiology.

[13]  D F Ollis,et al.  Kinetics of growth of the hydrogen‐oxidizing bacterium Alcaligenes eutrophus (ATCC 17707) in chemostat culture , 1984, Biotechnology and bioengineering.

[14]  H. Abe,et al.  Microbial synthesis and characterization of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) , 1995 .

[15]  W. Babel,et al.  Approaches to increase the economy of the PHB production , 1998 .

[16]  L. Matyska,et al.  Kinetics of d-3-hydroxybutyrate dehydrogenase from Paracoccus denitrificans , 1986 .

[17]  Hans G. Schlegel,et al.  β-Ketothiolase from Hydrogenomonas eutropha H16 and its significance in the regulation of poly-β-hydroxybutyrate metabolism , 1973 .

[18]  J. Merrick,et al.  Metabolism of poly-beta-hydroxybutyrate. II. Enzymatic synthesis of D-(-)-beta hydroxybutyryl coenzyme A by an enoyl hydrase from Rhodospirillum rubrum. , 1969, Biochemistry.

[19]  G. A. Ritchie,et al.  The purification and characterization of acetoacetyl-coenzyme A reductase from Azotobacter beijerinckii. , 1971, The Biochemical journal.

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

[21]  Fukui Tetsuya,et al.  Purification and characterization of NADP-linked acetoacetyl-CoA reductase from Zoogloea ramigera I-16-M. , 1987 .

[22]  A. Steinbüchel,et al.  Biochemical and genetic analysis of PHA synthases and other proteins required for PHA synthesis. , 1999, International journal of biological macromolecules.

[23]  Y. Doi,et al.  Biosynthesis of an unusual copolyester (10 mol % 3-hydroxybutyrate and 90 mol % 3-hydroxyvalerate units) in Alcaligenes eutrophus from pentanoic acid , 1987 .

[24]  E. Dawes,et al.  The role and regulation of energy reserve polymers in micro-organisms. , 1973, Advances in microbial physiology.

[25]  D. Byrom,et al.  Polymer synthesis by microorganisms: technology and economics , 1987 .

[26]  M. Ito,et al.  Purification and characterization of acetoacetyl-CoA synthetase from Zoogloea ramigera I-16-M. , 1982, European journal of biochemistry.

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

[28]  W. Page,et al.  Hyperproduction of Poly-β-Hydroxybutyrate during Exponential Growth of Azotobacter vinelandii UWD , 1989, Applied and environmental microbiology.

[29]  A. Steinbüchel,et al.  Cloning and nucleotide sequences of genes relevant for biosynthesis of poly(3-hydroxybutyric acid) in Chromatium vinosum strain D. , 1992, European journal of biochemistry.

[30]  N. Carr,et al.  Activation by Hg2+ of acetoacetyl‐CoA reductase in extracts of Rhodopseudomonas spheroides and Rhodomicrobium vannielii , 1969, FEBS letters.

[31]  B. Witholt,et al.  Polymerase C1 levels and poly(R-3-hydroxyalkanoate) synthesis in wild-type and recombinant Pseudomonas strains , 1997, Journal of bacteriology.

[32]  J. Merrick,et al.  Enzymatic Synthesis of Poly-β-Hydroxybutyric Acid in Bacteria , 1961, Nature.

[33]  Y. Okon,et al.  Production of the reserve material poly-β-hydroxybutyrate and its function in Azospirillum brasilense Cd , 1985 .

[34]  W. Babel,et al.  The auxiliary substrate concept — an approach for overcoming limits of microbial performances , 1993 .

[35]  J. Kingma,et al.  Characterization of intracellular inclusions formed by Pseudomonas oleovorans during growth on octane , 1983, Journal of bacteriology.

[36]  C. Anthony,et al.  The Biochemistry of Methylotrophs , 1982 .

[37]  G. W. Haywood,et al.  The role of NADH- and NADPH-linked acetoacetyl-CoA reductases in the poly-3-hydroxybutyrate synthesizing organism Alcaligenes eutrophus , 1988 .

[38]  G. Braunegg,et al.  Zur Kinetik des Wachstums und der Speicherung von Poly‐D(−)‐3‐hydroxybuttersäure bei Alcaligenes latus , 1985 .

[39]  Pj Piet Lemstra,et al.  Crystallization phenomena in bacterial poly[(R)-3-hydroxybutyrate]: 2. Embrittlement and rejuvenation , 1993 .

[40]  A. P. Sokolov,et al.  Purification and characterization of NADPH-dependent acetoacetyl-CoA reductase from Methylobacterium extorquens , 1997 .

[41]  J. P. van Dijken,et al.  Growth yields of microorganisms on methanol and methane. A theoretical study , 1975 .

[42]  A. Steinbüchel,et al.  Identification and characterization of two Alcaligenes eutrophus gene loci relevant to the poly(beta-hydroxybutyric acid)-leaky phenotype which exhibit homology to ptsH and ptsI of Escherichia coli , 1991, Journal of bacteriology.

[43]  C. Jones,et al.  Poly-3-hydroxybutyrate production by washed cells of Alcaligenes eutrophus; purification, characterisation and potential regulatory role of citrate synthase , 1997, Archives of Microbiology.

[44]  D. Jendrossek,et al.  Biodegradation of polyhydroxyalkanoic acids , 1996, Applied Microbiology and Biotechnology.

[45]  G. W. Haywood,et al.  Accumulation of a Polyhydroxyalkanoate Containing Primarily 3-Hydroxydecanoate from Simple Carbohydrate Substrates by Pseudomonas sp. Strain NCIMB 40135 , 1990, Applied and environmental microbiology.

[46]  J. Wilkinson,et al.  Poly-beta-hyroxybutyrate metabolism in washed suspensions of Bacillus cereus and Bacillus megaterium. , 1958, Journal of general microbiology.

[47]  S. Lee,et al.  High cell density culture of metabolically engineered Escherichia coli for the production of poly(3-hydroxybutyrate) in a defined medium. , 1998, Biotechnology and bioengineering.

[48]  Yong‐Hyun Lee,et al.  Metabolic characteristics of isocitrate dehydrogenase leaky mutant of Alcaligenes eutrophus and its utilization for poly-β-hydroxybutyrate production , 1996 .

[49]  W. Babel,et al.  Energy and reducing equivalent potential of C2-compounds for microbial growth† , 1988 .

[50]  C. Chavarie,et al.  Production of poly-(beta-hydroxybutyric-co-beta-hydroxyvaleric) acids , 1990, Applied and environmental microbiology.

[51]  T. Gerngross,et al.  Enzyme-catalyzed synthesis of poly[(R)-(-)-3-hydroxybutyrate]: formation of macroscopic granules in vitro. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[52]  J. Knowles Development of a natural degradable polymer for orthopaedic use. , 1993, Journal of medical engineering & technology.

[53]  Edwin A. Dawes,et al.  Characterization of two 3-ketothiolases possessing differing substrate specificities in the polyhydroxyalkanoate synthesizing organism Alcaligenes eutrophus , 1988 .

[54]  G. Huisman,et al.  Formation of Polyesters by Pseudomonas oleovorans: Effect of Substrates on Formation and Composition of Poly-(R)-3-Hydroxyalkanoates and Poly-(R)-3-Hydroxyalkenoates , 1988, Applied and environmental microbiology.

[55]  D. Seebach,et al.  Detection, synthesis, structure, and function of oligo(3-hydroxyalkanoates): contributions by synthetic organic chemists. , 1999, International journal of biological macromolecules.

[56]  S. Lee,et al.  Factors affecting the economics of polyhydroxyalkanoate production by bacterial fermentation , 1999, Applied Microbiology and Biotechnology.

[57]  A. Steinbüchel,et al.  Metabolic pathway for biosynthesis of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) from 4-hydroxybutyrate by Alcaligenes eutrophus. , 1995, European journal of biochemistry.

[58]  A. Sinskey,et al.  PHA synthase activity controls the molecular weight and polydispersity of polyhydroxybutyrate in vivo , 1997, Nature Biotechnology.

[59]  Y. Doi,et al.  Biosynthesis of copolyesters in Alcaligenes eutrophus H16 from carbon-13 labeled acetate and propionate , 1987 .

[60]  Y. W. Lee,et al.  Increased PHB productivity by high‐cell‐density fed‐batch culture of Alcaligenes latus, a growth‐associated PHB producer , 1996, Biotechnology and bioengineering.

[61]  W. Page,et al.  Control of polyhydroxyalkanoate synthesis in Azotobacter vinelandii strain UWD , 1994 .

[62]  G. A. Ritchie,et al.  The role of oxygen limitation in the formation of poly- -hydroxybutyrate during batch and continuous culture of Azotobacter beijerinckii. , 1972, The Biochemical journal.

[63]  E. Dawes,et al.  The regulation of poly-β-hydroxybutyrate metabolism in Azotobacter beijerinckii , 1973 .

[64]  G. Huisman,et al.  Pseudomonas putida KT2442 cultivated on glucose accumulates poly(3-hydroxyalkanoates) consisting of saturated and unsaturated monomers , 1992, Applied and environmental microbiology.

[65]  W. Babel Pecularities of methylotrophs concerning overflow metabolism, especially the synthesis of polyhydroxyalkanoates , 1992 .

[66]  H. Chang,et al.  Production of poly(3-hydroxybutyric acid) by recombinant Escherichia coli strains: genetic and fermentation studies. , 1995, Canadian journal of microbiology.

[67]  W. Babel Some theoretical considerations on overflow production of metabolites , 1986 .

[68]  W. Babel,et al.  Growth-associated synthesis of poly(hydroxybutyric acid) in Methylobacterium rhodesianum as an expression of an internal bottleneck , 1997, Applied Microbiology and Biotechnology.

[69]  C. Föllner,et al.  Considerations on the structure and biochemistry of bacterial polyhydroxyalkanoic acid inclusions. , 1995, Canadian journal of microbiology.

[70]  R. Bayly,et al.  Control of catechol meta-cleavage pathway in Alcaligenes eutrophus , 1983, Journal of bacteriology.

[71]  Huidong Shi,et al.  Metabolic Flux Analysis for Biosynthesis of Poly (β-Hydroxybutyric Acid) in Alcaligenes eutrophus from Various Carbon Sources , 1997 .

[72]  S. Lee,et al.  Cloning of the Alcaligenes latus Polyhydroxyalkanoate Biosynthesis Genes and Use of These Genes for Enhanced Production of Poly(3-hydroxybutyrate) in Escherichia coli , 1998, Applied and Environmental Microbiology.

[73]  G. Gottschalk,et al.  Formation and Utilization of Poly-β-Hydroxybutyric Acid by Knallgas Bacteria (Hydrogenomonas) , 1961, Nature.

[74]  H. Chang,et al.  Production of poly(3-hydroxybutyrate) by high cell density fed-batch culture of Alcaligenes eutrophus with phospate limitation. , 1997, Biotechnology and bioengineering.

[75]  G. W. Haywood,et al.  The importance of PHB-synthase substrate specificity in polyhydroxyalkanoate synthesis by Alcaligenes eutrophus , 1989 .

[76]  W. Babel The mixed substrate concept, applied for microbial syntheses of metabolites. , 1990, Biotechnology advances.

[77]  N. Lindley,et al.  Carbon and energy flux constraints in continuous cultures of Alcaligenes eutrophus grown on phenol. , 1998, Microbiology.

[78]  G. Huisman,et al.  Synthesis of poly-3-hydroxyalkanoates is a common feature of fluorescent pseudomonads , 1989, Applied and environmental microbiology.

[79]  Sang Yup Lee,et al.  Effect of fermentation performance on the economics of poly(3-hydroxybutyrate) production byAlcaligenes latus , 1998 .