title: Protamylasse, a residual compound of industrial starch production, provides a suitable medium for large-scale cyanophycin production.
abstract: Protamylasse is a residual compound occurring during the industrial production of starch from potatoes. It contains a variety of nutrients and all necessary minerals and could be used as a carbon, nitrogen, and energy source for the growth of bacteria and also for cyanophycin (CGP) biosynthesis. Media containing protamylasse as the sole compound diluted only in water were therefore examined for their suitability in CGP production. Among various bacterial strains investigated in this study, a recombinant strain of Escherichia coli DH1 harboring plasmid pMa/c5-914::cphA6803, which carries the cyanophycin synthetase structural gene (cphA) from Synechocystis sp. strain PCC6803, was found to be most suitable. Various cultivation conditions for high CGP contents were first optimized in shake flask cultures. The optimized conditions were then successfully applied to 30- and 500-liter fermentation scales in stirred tank reactors. A maximum CGP content of 28% (wt/wt) CGP per cell dry matter was obtained in 6% (vol/vol) protamylasse medium at an initial pH of 7.0 within a cultivation period of only 24 h. The CGP contents obtained with this recombinant strain employing protamylasse medium were higher than those obtained with the same strain cultivated in mineral salts medium or in expensive commercial complex media such as Luria-Bertani or Terrific broth. It was shown that most amino acids present in the protamylasse medium were almost completely utilized by the cells during cultivation. Exceptions were alanine, tryptophan, tyrosine, and most interestingly, arginine. Furthermore, CGP was easily isolated from protamylasse-grown cells by applying the acid extraction method. The CGP exhibited a molecular mass of about 26 to 30 kDa and was composed of 50% (mol/mol) aspartate, 46% (mol/mol) arginine, and 4% (mol/mol) lysine. The use of cheap residual protamylasse could contribute in establishing an economically and also ecologically feasible process for the biotechnological production of CGP.
authors.affiliation: Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität, Corrensstrasse 3, D-48149 Münster, Germany.
authors.forename: Yasser
authors.initials: Y
authors.lastname: Elbahloul
authors.forename: Kay
authors.initials: K
authors.lastname: Frey
authors.forename: Johan
authors.initials: J
authors.lastname: Sanders
authors.forename: Alexander
authors.initials: A
authors.lastname: Steinbüchel
mesh_terms: D000596:Amino Acids / Q000032:analysis / Q000378:metabolism*; D000681:Amylases / Q000737:chemistry / Q000378:metabolism*; D001426:Bacterial Proteins; D002241:Carbohydrates; D002264:Carboxylic Acids / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D005285:Fermentation; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D010940:Plant Proteins / Q000096:biosynthesis*; D010957:Plasmids; D013213:Starch
chemical_list: D000596:Amino Acids; D001426:Bacterial Proteins; D002241:Carbohydrates; D002264:Carboxylic Acids; D010940:Plant Proteins; C024250:cyanophycin; C507509:protamylasse; D013213:Starch; D000681:Amylases