Features of the biotechnologically relevant polyamide family “cyanophycins” and their biosynthesis in prokaryotes and eukaryotes

Abstract Cyanophycin, inclusions in cyanobacteria discovered by the Italian scientist Borzi in 1887, were characterized as a polyamide consisting of aspartic acid and arginine. Its synthesis in cyanobacteria was analyzed regarding growth conditions, responsible gene product, requirements, polymer structure and properties. Heterologous expression of diverse cyanophycin synthetases (CphA) in Escherichia coli enabled further enzyme characterization. Cyanophycin is a polyamide with variable composition and physiochemical properties dependent on host and cultivation conditions in contrast to the extracellular polyamides poly-γ-glutamic acid and poly-ε-l-lysine. Furthermore, recombinant prokaryotes and transgenic eukaryotes, including plants expressing different cphA genes, were characterized as suitable for production of insoluble cyanophycin regarding higher yields and modified composition for other requirements and applications. In addition, cyanophycin was characterized as a source for the synthesis of polyaspartic acid or N-containing bulk chemicals and dipeptides upon chemical treatment or degradation by cyanophycinases, respectively. Moreover, water-soluble cyanophycin derivatives with altered amino acid composition were isolated from transgenic plants, yeasts and recombinant bacteria. Thereby, the range of dipeptides could be extended by biological processes and by chemical modification, thus increasing the range of applications for cyanophycin and its dipeptides, including agriculture, food supplementations, medical and cosmetic purposes, synthesis of the polyacrylate substitute poly(aspartic acid) and other applications.

[1]  A. Steinbüchel,et al.  Guanidination of Soluble Lysine-Rich Cyanophycin Yields a Homoarginine-Containing Polyamide , 2014, Applied and Environmental Microbiology.

[2]  A. Steinbüchel,et al.  Solubility Behavior of Cyanophycin Depending on Lysine Content , 2013, Applied and Environmental Microbiology.

[3]  Alexander Steinb�chel,et al.  Increased Lysine Content Is the Main Characteristic of the Soluble Form of the Polyamide Cyanophycin Synthesized by Recombinant Escherichia coli , 2013, Applied and Environmental Microbiology.

[4]  A. Steinbüchel,et al.  Investigations on three genes in Ralstonia eutropha H16 encoding putative cyanophycin metabolizing enzymes , 2013, Applied Microbiology and Biotechnology.

[5]  W. Tseng,et al.  Assessments of growth conditions on the production of cyanophycin by recombinant Escherichia coli strains expressing cyanophycin synthetase gene , 2012, Biotechnology progress.

[6]  Sara M. Santos,et al.  Biomedical applications of dipeptides and tripeptides , 2012, Biopolymers.

[7]  J. Sanders,et al.  Production of cyanophycin in Rhizopus oryzae through the expression of a cyanophycin synthetase encoding gene , 2011, Applied Microbiology and Biotechnology.

[8]  David Hasson,et al.  State of the Art of Friendly “Green” Scale Control Inhibitors: A Review Article , 2011 .

[9]  A. Steinbüchel,et al.  Synthesis of a citrulline-rich cyanophycin by use of Pseudomonas putida ATCC 4359 , 2011, Applied Microbiology and Biotechnology.

[10]  A. Steinbüchel,et al.  Heterologous expression of Anabaena sp. PCC7120 cyanophycin metabolism genes cphA1 and cphB1 in Sinorhizobium (Ensifer) meliloti 1021 , 2011, Applied Microbiology and Biotechnology.

[11]  A. Steinbüchel,et al.  A novel plasmid addiction system for large-scale production of cyanophycin in Escherichia coli using mineral salts medium , 2011, Applied Microbiology and Biotechnology.

[12]  A. Steinbüchel,et al.  Dipeptides in nutrition and therapy: cyanophycin-derived dipeptides as natural alternatives and their biotechnological production , 2010, Applied Microbiology and Biotechnology.

[13]  A. Steinbüchel,et al.  Establishment of a simple and effective isolation method for cyanophycin from recombinant Saccharomyces cerevisiae , 2010, Applied Microbiology and Biotechnology.

[14]  G. Wei,et al.  Biological flocculation treatment on distillery wastewater and recirculation of wastewater. , 2009, Journal of hazardous materials.

[15]  A. Steinbüchel,et al.  Establishment of Cyanophycin Biosynthesis in Pichia pastoris and Optimization by Use of Engineered Cyanophycin Synthetases , 2009, Applied and Environmental Microbiology.

[16]  W. Lockau,et al.  Tuber-specific cphA expression to enhance cyanophycin production in potatoes. , 2009, Plant biotechnology journal.

[17]  M. J. Blish.  On the Chemical Constitution of the Proteins of Wheat Flour and Its Relation to Baking Strength , 2009 .

[18]  A. Steinbüchel,et al.  Metabolic Engineering of Saccharomyces cerevisiae for Production of Novel Cyanophycins with an Extended Range of Constituent Amino Acids , 2009, Applied and Environmental Microbiology.

[19]  J. Suh,et al.  The role of the C-terminal region of cyanophycin synthetase from Nostoc ellipsosporum NE1 in its enzymatic activity and thermostability: a key function of Glu(856). , 2009, Biochimica et biophysica acta.

[20]  A. Steinbüchel,et al.  Biotechnological Process for Production of β-Dipeptides from Cyanophycin on a Technical Scale and Its Optimization , 2008, Applied and Environmental Microbiology.

[21]  K. Kino,et al.  A cyanophycin synthetase from Thermosynechococcus elongatus BP-1 catalyzes primer-independent cyanophycin synthesis , 2008, Applied Microbiology and Biotechnology.

[22]  W. Lockau,et al.  Plastid targeting strategies for cyanophycin synthetase to achieve high-level polymer accumulation in Nicotiana tabacum. , 2008, Plant biotechnology journal.

[23]  A. Steinbüchel,et al.  Anaerobic and Aerobic Degradation of Cyanophycin by the Denitrifying Bacterium Pseudomonas alcaligenes Strain DIP1 and Role of Three Other Coisolates in a Mixed Bacterial Consortium , 2008, Applied and Environmental Microbiology.

[24]  A. Steinbüchel,et al.  Synthesis and Accumulation of Cyanophycin in Transgenic Strains of Saccharomyces cerevisiae , 2008, Applied and Environmental Microbiology.

[25]  J. Khattar,et al.  Nutrient stress causes akinete differentiation in cyanobacteriumAnabaena torulosa with concomitant increase in nitrogen reserve substances , 2008, Folia Microbiologica.

[26]  Alexander Steinbüchel,et al.  Assessment of technological options and economical feasibility for cyanophycin biopolymer and high-value amino acid production , 2007, Applied Microbiology and Biotechnology.

[27]  Hans Mooibroek,et al.  Bio-refinery as the bio-inspired process to bulk chemicals. , 2007, Macromolecular bioscience.

[28]  M. Fischbach,et al.  Assembly-line enzymology for polyketide and nonribosomal Peptide antibiotics: logic, machinery, and mechanisms. , 2006, Chemical reviews.

[29]  Kim Kye-seong,et al.  Establishment of a simple and effective method for isolating male germline stem cells (GSCs) from testicular cells of neonatal and adult mice , 2006 .

[30]  A. Steinbüchel,et al.  Application of a KDPG-aldolase gene-dependent addiction system for enhanced production of cyanophycin in Ralstonia eutropha strain H16. , 2006, Metabolic engineering.

[31]  J. Sanders,et al.  Protamylasse, a Residual Compound of Industrial Starch Production, Provides a Suitable Medium for Large-Scale Cyanophycin Production , 2005, Applied and Environmental Microbiology.

[32]  U. Conrad.  Polymers from plants to develop biodegradable plastics. , 2005, Trends in plant science.

[33]  A. Steinbüchel,et al.  Degradation of Cyanophycin by Sedimentibacter hongkongensis Strain KI and Citrobacter amalonaticus Strain G Isolated from an Anaerobic Bacterial Consortium , 2005, Applied and Environmental Microbiology.

[34]  A. Steinbüchel,et al.  Investigations on the solubility behavior of cyanophycin. Solubility of cyanophycin in solutions of simple inorganic salts. , 2005, Biomacromolecules.

[35]  W. Lockau,et al.  Production of cyanophycin, a suitable source for the biodegradable polymer polyaspartate, in transgenic plants. , 2005, Plant biotechnology journal.

[36]  A. Steinbüchel,et al.  Physiological Conditions Conducive to High Cyanophycin Content in Biomass of Acinetobacter calcoaceticus Strain ADP1 , 2005, Applied and Environmental Microbiology.

[37]  A. Steinbüchel,et al.  Partial purification and characterization of a non-cyanobacterial cyanophycin synthetase from Acinetobacter calcoaceticus strain ADP1 with regard to substrate specificity, substrate affinity and binding to cyanophycin. , 2004, Microbiology.

[38]  A. Steinbüchel,et al.  Identification of the Anabaena sp. strain PCC7120 cyanophycin synthetase as suitable enzyme for production of cyanophycin in gram-negative bacteria like Pseudomonas putida and Ralstonia eutropha. , 2004, Biomacromolecules.

[39]  W. Zimmerman,et al.  Cyanophycin granule polypeptide protease in a unicellular cyanobacterium , 1984, Archives of Microbiology.

[40]  R. Simon.  The effect of chloramphenicol on the production of cyanophycin granule polypeptide in the blue-green alga Anabaena cylindrica , 2004, Archiv für Mikrobiologie.

[41]  A. Steinbüchel,et al.  Isolation and characterization of gram-positive cyanophycin-degrading bacteria-kinetic studies on cyanophycin depolymerase activity in aerobic bacteria. , 2004, Biomacromolecules.

[42]  M. M. Allen,et al.  Variations in the amino acid composition of cyanophycin in the cyanobacterium Synechocystis sp. PCC 6308 as a function of growth conditions , 2004, Archives of Microbiology.

[43]  Alexander Steinbüchel,et al.  Isolation of Cyanophycin-degrading Bacteria, Cloning and Characterization of an Extracellular Cyanophycinase Gene (cphE) from Pseudomonas anguilliseptica Strain BI , 2002, The Journal of Biological Chemistry.

[44]  A. Steinbüchel,et al.  Technical-Scale Production of Cyanophycin with Recombinant Strains of Escherichia coli , 2002, Applied and Environmental Microbiology.

[45]  W. Lockau,et al.  Cyanophycin Synthetase-Like Enzymes of Non-Cyanobacterial Eubacteria: Characterization of the Polymer Produced by a Recombinant Synthetase of Desulfitobacterium hafniense , 2002, Zeitschrift fur Naturforschung. C, Journal of biosciences.

[46]  Scott R. Miller,et al.  Cyanophycin Production in a Phycoerythrin-Containing Marine Synechococcus Strain of Unusual Phylogenetic Affinity , 2002, Applied and Environmental Microbiology.

[47]  Alexander Steinbüchel,et al.  Evaluation of non-cyanobacterial genome sequences for occurrence of genes encoding proteins homologous to cyanophycin synthetase and cloning of an active cyanophycin synthetase from Acinetobacter sp. strain DSM 587 , 2002, Archives of Microbiology.

[48]  A. Steinbüchel,et al.  Molecular Characterization of a Thermostable Cyanophycin Synthetase from the Thermophilic Cyanobacterium Synechococcus sp. Strain MA19 and In Vitro Synthesis of Cyanophycin and Related Polyamides , 2002, Applied and Environmental Microbiology.

[49]  이주희,et al.  Preparation and Swelling Behavior of Biodegradable Superabsorbent Gels Based on Polyaspartic Acid , 2002 .

[50]  A. Steinbüchel,et al.  Purification of Synechocystis sp. Strain PCC6308 Cyanophycin Synthetase and Its Characterization with Respect to Substrate and Primer Specificity , 2001, Applied and Environmental Microbiology.

[51]  A. Steinbüchel,et al.  Heterologous expression of cyanophycin synthetase and cyanophycin synthesis in the industrial relevant bacteria Corynebacterium glutamicum and Ralstonia eutropha and in Pseudomonas putida. , 2001, Biomacromolecules.

[52]  A. Steinbüchel,et al.  Molecular characterization of the cyanophycin synthetase from Synechocystis sp. strain PCC6308 , 2000, Archives of Microbiology.

[53]  K. Baier,et al.  Biosynthesis of the cyanobacterial reserve polymer multi-L-arginyl-poly-L-aspartic acid (cyanophycin): mechanism of the cyanophycin synthetase reaction studied with synthetic primers. , 2000, European journal of biochemistry.

[54]  A. Steinbüchel,et al.  Axenic cultivation of anoxygenic phototrophic bacteria, cyanobacteria, and microalgae in a new closed tubular glass photobioreactor , 2000, Applied Microbiology and Biotechnology.

[55]  A. Steinbüchel,et al.  Purification and characterization of cyanophycin and cyanophycin synthetase from the thermophilic Synechococcus sp. MA19. , 1999, FEMS microbiology letters.

[56]  W. Lockau,et al.  Cyanophycinase, a peptidase degrading the cyanobacterial reserve material multi-L-arginyl-poly-L-aspartic acid (cyanophycin): molecular cloning of the gene of Synechocystis sp. PCC 6803, expression in Escherichia coli, and biochemical characterization of the purified enzyme. , 1999, European journal of biochemistry.

[57]  F. Leganés,et al.  A transposition-induced mutant of Nostoc ellipsosporum implicates an arginine-biosynthetic gene in the formation of cyanophycin granules and of functional heterocysts and akinetes. , 1998, Microbiology.

[58]  E. Baker,et al.  Structural homologies with ATP- and folate-binding enzymes in the crystal structure of folylpolyglutamate synthetase. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[59]  W. Lockau,et al.  Molecular characterization of cyanophycin synthetase, the enzyme catalyzing the biosynthesis of the cyanobacterial reserve material multi-L-arginyl-poly-L-aspartate (cyanophycin). , 1998, European journal of biochemistry.

[60]  M. Page-Sharp,et al.  Cyanophycin and glycogen synthesis in a cyanobacterial Scytonema species in response to salt stress , 1998 .

[61]  M. Schwamborn.  Chemical synthesis of polyaspartates: a biodegradable alternative to currently used polycar☐ylate homo- and copolymers , 1998 .

[62]  W. Joentgen,et al.  Chemical synthesis of polyaspartates. a biodegradable alternative to currently used polycarboxylate homo- and copolymers , 1998 .

[63]  Michael Y. Galperin,et al.  A diverse superfamily of enzymes with ATP‐dependent carboxylate—amine/thiol ligase activity , 1997, Protein science : a publication of the Protein Society.

[64]  D. Pompliano,et al.  Conditionally lethal Escherichia coli murein mutants contain point defects that map to regions conserved among murein and folyl poly-gamma-glutamate ligases: identification of a ligase superfamily. , 1997, Biochemistry.

[65]  V. Ravindran,et al.  Investigations on the Guanidination of Lysine in Proteins , 1996 .

[66]  Toshio Fukui,et al.  Structure of the multifunctional loops in the nonclassical ATP-binding fold of glutathione synthetase , 1996, Nature Structural Biology.

[67]  J. Meredith,et al.  Visceral protein levels in trauma patients are greater with peptide diet than with intact protein diet. , 1990, The Journal of trauma.

[68]  M. Wyman,et al.  Absence of the nitrogen reserve polymer cyanophycin from marine Synechococcus species , 1987 .

[69]  M. Gupta,et al.  Enzyme Activities Related to Cyanophycin Metabolism in Heterocysts and Vegetative Cells of Anabaena spp. , 1981 .

[70]  S. Stevens,et al.  Accumulation of Cyanophycin Granules as a Result of Phosphate Limitation in Agmenellum quadruplicatum. , 1981, Plant physiology.

[71]  G. Mclendon,et al.  Structural characterization of the cyanophycin granule polypeptide of Anabaena cylindrica by circular dichroism and Raman spectroscopy. , 1980, Biochimica et biophysica acta.

[72]  P. Weathers,et al.  Cyanophycin granule polypeptide formation and degradation in the cyanobacterium Aphanocapsa 6308 , 1980, Journal of bacteriology.

[73]  P. Weathers,et al.  Structure and composition of cyanophycin granules in the cyanobacterium Aphanocapsa 6308 , 1980, Journal of bacteriology.

[74]  R. Simon.  The biosynthesis of multi-L-arginyl-poly(L-aspartic acid) in the filamentous cyanobacterium Anabaena cylindrica. , 1976, Biochimica et biophysica acta.

[75]  R. Simon.  Measurement of the Cyanophycin Granule Polypeptide Contained in the Blue-Green Alga Anabaena cylindrica , 1973, Journal of bacteriology.

[76]  D. Perrett,et al.  Dipeptide absorption in man , 1972, Gut.

[77]  R. Simon.  Cyanophycin Granules from the Blue-Green Alga Anabaena cylindrica: A Reserve Material Consisting of Copolymers of Aspartic Acid and Arginine. , 1971, Proceedings of the National Academy of Sciences of the United States of America.

[78]  L. Messineo.  Modification of the Sakaguchi reaction: Spectrophotometric determination of arginine in proteins without previous hydrolysis , 1966 .