Submerged Production, Purification, and Crystallization of Acid Carboxypeptidase from Penicillium janthinellum IFO-8070

Sadaji Yokoyama, Akira Oobayashi, Osamu Tanabe, Eiji Ichishima
1974 Applied microbiology  
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% KH2PO; 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-molecularweight acid carboxypeptidase were produced. Acid
more » ... 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.
doi:10.1128/aem.28.5.742-747.1974 fatcat:j7vhfbdtfjbe7i2wqekxrwddxa