title: Establishment of a simple and effective isolation method for cyanophycin from recombinant Saccharomyces cerevisiae.
abstract: An efficient, time-saving, and cost-effective method for isolation of the polyamide cyanophycin from recombinant Saccharomyces cerevisiae was established. Due to its simple procedure, this isolation method may be also applicable at industrial scale and also to other intracellular compounds in this yeast. Production of cyanophycin gained preferential interest in the past, as degradation products thereof are of pharmaceutical and technical interest. Recently, it was shown that Saccharomyces cerevisiae represents a putative candidate for cyanophycin synthesis at industrial scale. For identification of optimal isolation procedures, several parameters such as heat stress, freeze drying, and freeze/thaw cycles of transgenic yeast cells were compared for their effectiveness of cyanophycin isolation. Additionally, optimal resuspension solutions for the applied cells and minimal required materials or chemicals were determined to make the process most environmentally and economically friendly. Maximal cyanophycin granule polypeptide yields of 21% (w/w) were obtained after incubation of dry cells at 70 degrees C or 80 degrees C and precipitation of the polymer with two volumes of ethanol.
authors.affiliation: Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität, 48149 Münster, Germany.
authors.forename: Anna
authors.initials: A
authors.lastname: Steinle
authors.forename: Alexander
authors.initials: A
authors.lastname: Steinbüchel
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D001709:Biotechnology / Q000379:methods; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D000431:Ethanol / Q000378:metabolism; D005285:Fermentation; D005615:Freezing; D006358:Hot Temperature; D010453:Peptide Synthases / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D013312:Stress, Physiological; D013379:Substrate Specificity; D046939:Synechocystis / Q000201:enzymology / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; C024250:cyanophycin; D000431:Ethanol; D010453:Peptide Synthases; C113257:cyanophycin synthase, bacteria