Abstract
A rapid method for preparing cell-free extracts of Aspergillus ochraceus was developed. Mycelial mats were prefrozen in liquid nitrogen, ground to a fine powder in a cold mortar, and homogenized in an all-glass mechanical homogenizer. This method provided preparations averaging 43.0 mg of protein per g of mycelium (wet weight). The method was fast, efficient, and did not subject the extract to temperatures above 1 C or to heavy metals. The preparation method was suitable for studying a variety of in vitro fungal enzyme systems. Amylase, acid phosphatase, alkaline phosphatase, catalase, fatty acid synthetase, glucose-6-phosphate dehydrogenase, beta-glucosidase, beta fructofuranosidase, and trehalase activities were measurable in the preparations.
Full text
PDF
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- BEERS R. F., Jr, SIZER I. W. A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem. 1952 Mar;195(1):133–140. [PubMed] [Google Scholar]
- BERNFELD P. Enzymes of starch degradation and synthesis. Adv Enzymol Relat Subj Biochem. 1951;12:379–428. doi: 10.1002/9780470122570.ch7. [DOI] [PubMed] [Google Scholar]
- Davis N. D., Searcy J. W., Diener U. L. Production of ochratoxin A by Aspergillus ochraceus in a semisynthetic medium. Appl Microbiol. 1969 May;17(5):742–744. doi: 10.1128/am.17.5.742-744.1969. [DOI] [PMC free article] [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]
- STINE G. J., STRICKLAND W. N., BARRATT R. W. METHODS OF PROTEIN EXTRACTION FROM NEUROSPORA CRASSA. Can J Microbiol. 1964 Feb;10:29–35. doi: 10.1139/m64-005. [DOI] [PubMed] [Google Scholar]
- Scott D. B. Toxigenic fungi isolated from cereal and legume products. Mycopathol Mycol Appl. 1965 Apr 14;25(3):213–222. doi: 10.1007/BF02049914. [DOI] [PubMed] [Google Scholar]
- van der Merwe K. J., Steyn P. S., Fourie L., Scott D. B., Theron J. J. Ochratoxin A, a toxic metabolite produced by Aspergillus ochraceus Wilh. Nature. 1965 Mar 13;205(976):1112–1113. doi: 10.1038/2051112a0. [DOI] [PubMed] [Google Scholar]