Abstract
The extracellular proteinases of Aspergillus oryzae EI 212 were separated into two active fractions by (NH4)2SO4 and ethanol fractionation followed by diethyl-aminoethyl-Sephadex A-50 and hydroxyapatite chromatography. The molecular weight was estimated by gel filtration to be about 70,000 and 35,000 for proteinases I and II, respectively. Optimum pH for casein and hemoglobin hydrolysis was 6.5 at 60 C for proteinase I and 10.0 at 45 C for proteinase II, and for gelatin hydrolysis it was 6.5 at 45 C for both enzymes. The enzymes were stable over the pH range 6 to 8 at 30 C for 60 min. The enzyme activity for both the proteinases was accelerated by Cu2+ and inhibited by Fe2+, Fe3+, Hg2+, and Ag+. Halogenators (e.g., N-chlorosuccinimide) and diisopropyl fluorophosphate inhibited proteinase II. Sulfhydryl reagents such as p-chloromercuribenzoate and iodoacetate inhibited proteinase I. Sulfhydryl compounds accelerated the action of both enzymes.
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Selected References
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- Andrews P. Estimation of the molecular weights of proteins by Sephadex gel-filtration. Biochem J. 1964 May;91(2):222–233. doi: 10.1042/bj0910222. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Christison J., Martin S. M. Isolation and preliminary characterization of an extracellular protease of Cytophaga sp. Can J Microbiol. 1971 Sep;17(9):1207–1216. doi: 10.1139/m71-193. [DOI] [PubMed] [Google Scholar]
- DAVIS N. C., SMITH E. L. Assay of proteolytic enzymes. Methods Biochem Anal. 1955;2:215–257. doi: 10.1002/9780470110188.ch8. [DOI] [PubMed] [Google Scholar]
- HJERTEN S., LEVIN O., TISELIUS A. Protein chromatography on calcium phosphate columns. Arch Biochem Biophys. 1956 Nov;65(1):132–155. doi: 10.1016/0003-9861(56)90183-7. [DOI] [PubMed] [Google Scholar]
- Han Y. W., Srinivasan V. R. Purification and characterization of beta-glucosidase of Alcaligenes faecalis. J Bacteriol. 1969 Dec;100(3):1355–1363. doi: 10.1128/jb.100.3.1355-1363.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kundu A. K., Das S., Manna S., Pal N. Extracellular proteinases of Aspergillus oryzae. Appl Microbiol. 1968 Nov;16(11):1799–1801. doi: 10.1128/am.16.11.1799-1801.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kundu A. K., Das S. Production of amylase in liquid culture by a strain of Aspergillus oryzae. Appl Microbiol. 1970 Apr;19(4):598–603. doi: 10.1128/am.19.4.598-603.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kundu A. K., Manna S., Pal N. Purification and properties of a new extracellular collagenase from Aspergillus sclerotiorum. Indian J Exp Biol. 1974 Sep;12(5):441–443. [PubMed] [Google Scholar]
- Kundu A. K., Manna S. Purification and properties of a new alpha-glucanase from Aspergillus oryzae. Indian J Exp Biol. 1971 Jan;9(1):75–78. [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Lysenkov N. V., Tsyperovich A. S. Vlastyvosti proteazy Aspergillus flavus. Ukr Biokhim Zh. 1969;41(2):157–162. [PubMed] [Google Scholar]
- MASSEY V. Studies on fumarase. III. The effect of temperature. Biochem J. 1953 Jan;53(1):72–79. doi: 10.1042/bj0530072. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nordwig A., Jahn W. F. A collagenolytic enzyme from Aspergillus oryzae. Purification and properties. Eur J Biochem. 1968 Feb;3(4):519–529. doi: 10.1111/j.1432-1033.1967.tb19562.x. [DOI] [PubMed] [Google Scholar]
- Nordwig A., Jahn W. F. Spezifitätseigenschaften einer Protease aus Aspergillus oryzae. Hoppe Seylers Z Physiol Chem. 1966;345(4):284–287. [PubMed] [Google Scholar]
- Oleniacz W. S., Pisano M. A. Proteinase production by a species of Cephalosporium. Appl Microbiol. 1968 Jan;16(1):90–96. doi: 10.1128/am.16.1.90-96.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Van Heyningen S., Secher D. S. A new alkaline protease from Acremonium kiliense. Biochem J. 1971 Dec;125(4):1159–1160. doi: 10.1042/bj1251159. [DOI] [PMC free article] [PubMed] [Google Scholar]