BIOCHEMICAL AND ELECTRON-MICROSCOPIC STUDIES OF THE STREPTOMYCES-RETICULI CELLULASE (AVICELASE) IN ITS MYCELIUM-ASSOCIATED AND EXTRACELLULAR FORMS
Autor(en): | SCHLOCHTERMEIER, A NIEMEYER, F SCHREMPF, H |
Stichwörter: | 1,4-BETA-D-GLUCAN CELLOBIOHYDROLASE; Biotechnology & Applied Microbiology; CELLULOLYTIC ENZYMES; CLOSTRIDIUM-THERMOCELLUM; COMPLEX; FAMILIES; GENES; Microbiology; PURIFICATION; STERCORARIUM; SYSTEM; TRICHODERMA-REESEI | Erscheinungsdatum: | 1992 | Herausgeber: | AMER SOC MICROBIOLOGY | Journal: | APPLIED AND ENVIRONMENTAL MICROBIOLOGY | Volumen: | 58 | Ausgabe: | 10 | Startseite: | 3240 | Seitenende: | 3248 | Zusammenfassung: | Streptomyces reticuli is able to grow efficiently with crystalline cellulose (Avicel) as the sole carbon source. Cultivation in the presence of the nonionic detergent Tween 80 at a concentration of 0.1% led to a 10-fold increase in extracellular cellulolytic activity. Under these conditions, one single 82-kDa cellulase (Avicelase) capable of degrading crystalline and soluble cellulose as well as cellodextrins and p-nitrophenylcellobioside was purified to apparent homogeneity by a procedure which consisted of two consecutive anion-exchange chromatographies followed by chromatofocusing. Aggregation, which was a major problem during protein purification, could be avoided by including Triton X-100 at a concentration of 0.1% in every chromatographic step. The Avicelase was identified in extracellular and mycelium-associated forms, the latter of which could be released efficiently by nonionic detergents. In addition, a 42-kDa truncated form retaining cellulolytic activity was identified which had been generated from the 82-kDa enzyme by a protease. Antibodies raised against the mycelium-associated Avicelase reacted with the 42-kDa derivative and the extracellular form. The mycelial association of the enzyme was confirmed by immunofluorescence and immunoelectron microscopies. |
ISSN: | 00992240 | DOI: | 10.1128/AEM.58.10.3240-3248.1992 |
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