Purification and characterization of cyanophycin and cyanophycin synthetase from the thermophilic Synechococcus sp. MA19.
The biosynthesis and accumulation of cyanophycin in the thermophilic cyanobacterium Synechococcus sp. MA19 were studied. By growing the cells in a 80-l closed tubular photobioreactor under controlled conditions, the cells accumulated cyanophycin amounting up to 3.5% of the dry cell matter. The cyanophycin was purified and chemical analysis showed that it was composed of arginine and aspartic acid occurring at a molar ratio of 1:0.9. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a broad distribution of the apparent molecular masses ranging from 20 to 130 kDa with a maximum at 50 kDa. During a three-step purification procedure involving ion exchange chromatography and gel filtration, the cyanophycin synthetase from strain MA19 was purified 144-fold to electrophoretic homogeneity. It consisted of only one single type of subunit exhibiting an apparent molecular mass of 130 kDa. The enzyme catalyzed the polymerization of arginine and aspartate at elevated temperatures and was even active at 80 degrees C.
Hai
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T
Institut für Mikrobiologie der Westfälischen Wilhelms-Universität, Corrensstrasse 3, 48149, Münster, Germany.
Oppermann-Sanio
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FB
Steinbüchel
A
A
D001120:Arginine / Q000378:metabolism; D001224:Aspartic Acid / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D019149:Bioreactors / Q000382:microbiology; D002851:Chromatography, High Pressure Liquid; D000458:Cyanobacteria / Q000737:chemistry* / Q000201:enzymology* / Q000254:growth & development; D004591:Electrophoresis, Polyacrylamide Gel; D010453:Peptide Synthases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification*; D013696:Temperature
D001426:Bacterial Proteins; D010940:Plant Proteins; C024250:cyanophycin; D001224:Aspartic Acid; D001120:Arginine; D010453:Peptide Synthases; C113257:cyanophycin synthase, bacteria