Abstract
The cellulase system ofBacillus circulans F-2 effectively hydrolyzed carboxymethyl cellulose (CMC), xylan, avicel, cellobiose, filter paper, cotton, andp-nitrophenyl-Β-D-cellobioside, and the crude enzyme produced mainly glucose from digestion of avicel. Two major and one minor peaks of enzyme activities were eluted on DEAE ion-exchange chromatography, and designated cellulase complex I(C-I) and complex II(C-II) for the two major peaks, and cellulase-III for a minor peak. C-I and C-II were further purified on gel filtration column of a TSK-Gel SW G3000 ×L. The molecular masses of C-I and C-II were estimated to be about 669 and 443 kDa, respectively. Sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis of the C-I and C-II complexes showed that the C-I complex was present as a multiple protein complex, consisting of at least five CMCases and two xylanases, and that the C-II complex was consisted of at least three CMCase and four xylan ases. C-I showed high activities of cellohydrolase, CMCase, xylanase, and Β-glucosidase, whereas C-II showed high activities of CMCase, xylanase, avicelase, and Β-glucosidase. The outstanding property of the C-II was its high hydrolytic activity toward filter paper, a highly resistant substrate against enzymatic degradation. However, cellulaseIII showed only strong avicelase activity. These results indicated that the cellulase system of the strain exists as multiple complex forms.
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Abbreviations
- CMC:
-
carboxylmethyl cellulose
- FP:
-
filter paper
- kDa:
-
kilodalton
- DNS:
-
3′,5′-dinitrosalicylic acid
- pNPC:
-
p-nitrophenyl-Β-d-cellobioside
- pNPX:
-
p-nitrophenyl-Β-d-xylopyranoside
- pNPG:
-
p- nitrophenyl-Β-d-glucopyranoside
- PAGE:
-
polyacrylamide gel electrophoresis
- CBH:
-
cellobiohydrolase
- SDS:
-
sodium dodecyl sulfate
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Kim, CH., Kim, DS. Extracellular cellulolytic enzymes ofBacillus circulans are present as two multipleprotein complexes. Appl Biochem Biotechnol 42, 83–94 (1993). https://doi.org/10.1007/BF02788904
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DOI: https://doi.org/10.1007/BF02788904