Submerged production, purification, and crystallization of acid carboxypeptidase from Penicillium janthinellum IFO-8070.
Penicillium janthinellum IFO-8070 produced an acid carboxypeptidase of molecular weight 51,000 in a liquid medium at 25 C. Maximum enzyme concentration was obtained within 3 to 6 days in a medium containing 2% wheat bran, 1% defatted soybean, and 1% KH(2)PO(4); the initial pH was 2 to 4. When submerged aerobic conditions were used, a 51,000-molecular-weight acid carboxypeptidase was produced and no detectable amounts of 160,000-molecular-weight acid carboxypeptidase were produced. Acid carboxypeptidase with a molecular weight of 51,000 was purified 330-fold from koji culture to yield a crystalline protein which was demonstrated by disc electrophoresis to be homogeneous. The purification method included ammonium sulfate fractionation, Amberlite CG-50 chromatography, acetone fractionation, Amberlite CG-50 rechromatography, and concentration in a collodion bag. The specific activity of the enzyme was about three times more than that of the acid carboxypeptidase from Aspergillus saitoi.
D000096:Acetone; D000332:Aerobiosis; D000645:Ammonium Sulfate; D002268:Carboxypeptidases / Q000096:biosynthesis; D011232:Chemical Precipitation; D002852:Chromatography, Ion Exchange; D003460:Crystallization; D003470:Culture Media; D004589:Electrophoresis, Disc; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D010407:Penicillium / Q000201:enzymology* / Q000254:growth & development; D010710:Phosphates; D013025:Glycine max; D014908:Triticum
D003470:Culture Media; D010710:Phosphates; D000096:Acetone; D002268:Carboxypeptidases; D000645:Ammonium Sulfate