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The production of exo‐β‐glucosidases from Penicillium spinulosum chemostat and liquid shake cultures
Author(s) -
Wheeler Timothy W,
Deffit Sarah N,
Kochen Michael A,
Anderson Ryan T,
Bonetti Sandra J
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.911.3
Subject(s) - cellulase , chemostat , chemistry , enzyme , food science , microbiology and biotechnology , biochemistry , cellulose , biology , bacteria , genetics
The objectives of this project were to: (1) determine optimal conditions for production of exo‐β‐glucosidases or cellulases in both chemostat and flask cultures of Penicillium and (2) isolate and characterize this activity from P. spinulosum . In P. spinulosum chemostat cultures, strains were selected by using cellulose as the sole carbon source and the resulting strains produced a fourfold increase in enzyme. In liquid shake cultures, trials were performed to optimize enzyme production by P. spinulosum in cultures grown on either glucose‐ or galactose‐containing standard growth media (Glc‐ or Gal‐SGM). Glc‐SGM was selected for enzyme production because cellulase activity peaked on days 12 and 13, remained consistently higher and contained fewer contaminating activities than in Gal‐SGM. Cellulase was purified from 13‐day Glc‐SGM culture filtrates by successive DEAE anion exchange and Sephacryl S‐300 size exclusion chromatographies. SDS‐PAGE was performed on isolated cellulase fractions to resolve a primary (~ 100 kDa) protein band for sequencing. The isolated cellulase was free of contaminating glycohydrolase activities. Research support provided by the Merck Institute for Science Education.