• Korean Journal of Microbiology
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• v.26 no.2
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• pp.122-128
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• 1988

The enzymatic properties of soil cellobiohydrolase were examined and compared with those of cellobiohydrolase-active extracts from soil in the forms of enzyme-humic complex and humicfree enzyme, and cellobiohydrolase partially pruified from Aspergillus niger. The pH optima of soil cellobiohydrolase and cellobiohydrolase-humic complex were greater by 1.5-3.0 pH units than those of cellobiohydrolase in humic-free extract and from A. niger. Soil cellobiohydrolase and cellobiohydrolase-humic complex were remarkably resistant to thermal denaturation and proteolysis. These results confirm that cellobiohydrolase in soil is atable in conditions which rapidly inactivate microbial cellobiohydrolase and that its stability is due to the immobilization of this enzyme by association with humic substances. The Michaelis-Menten constants (Km) for soil, cellobiohydrolase-humic complex, humic free extract and cellobiohydrolase from A. niger were 22.1mg/ml, 11.3mg/ml, 10.6mg/ml and 4.5 mg/ml of Avicel, respectively.

• Microbiology and Biotechnology Letters
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• v.18 no.4
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• pp.390-393
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• 1990

A cellobiohydrolase was purified from the culture broth of Trichoderma uiride by using immunoaffinity chromatography. A single protein band in polyacylamide gel electrophoresis and isoelectrofocusing after immunoaffinity purification corresponded to cellobiohydrolase activity. A immunoaffinity purified cellobiohydrolase is more effective in the hydrolysis of highly crystalline cellulose than amorphous cellulose.

• Proceedings of the Microbiological Society of Korea Conference
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• 1991.04a
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• pp.187-199
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• 1991

Penicillium verruculosum 배양액으로부터 소단위체 분자량이 60,000(cellobiohydrolase I)과 66,000(cellobiohydrolase II) 및 76,000(cellobiohydrolase III)인 cellobiohydrolase를 분리 정제하여 그들의 일반적 특성을 검토하였다. 이들 세 효소들은 모두 당단백질로서 cellobiohydrolase I, II 그리고 III는 각각 $8.6\%$, $4.2\%$ 그리고 $8.5\%$의 당함량을 나타냈으며 세 효소 모두 pH 4.5 - 5.0, 온도 $50 - 60^{\circ}C$에서 최적조건을 나타냈다. Cellobiohydrolase I, II 및 III는 모두 Avicel, cotton, 여지 등의 결정성 섬유소 뿐만 아니라 carboxymethyl cellulose에도 활성도를 나타냈다. 정제된 cellobiohydrolase I, II 및 III의 Avicel에 대한 비 활성도는 각각 0.07, 0.10, 그리고 0.07 unit per mg. of protein 이었으며 Avicel 분해 생성물은 거의 cellobiose였다. 또한 p-Nitrophenyl-$\beta$-D-cellobioside를 기질로 하였을 때 이들 세 효소 모두 포도당에 의해 활성도가 저해되지 않은 반면 cellobiose에 의해서는 저해되었다. 아미노산조성, 트립신에 의한 펩타이드들의 용출양상 그리고 항체를 이용한 Immunoblotting 결과로부터 cellobiohydrolase II와 III는 동일 유전자산물이거나 1차 구조가 거의 유사한 단백질로 추정된다. Cellobiohydrolase II로부터 분리한 2 개의 펩타이드의 아미노산 서열은 Trichoderma cellobiohydrolase I과 상동성을 보였으며 또한 이 두 호소의 아미노산조성은 매우 유사하였다.

• Microbiology and Biotechnology Letters
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• v.17 no.4
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• pp.365-369
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• 1989

Thermal inactivation of Tyichoderma viride cellobiohydrolase was investigated by immunoassay and residual enzyme assay such as carboxymethyl cellulase (CMCase) and filter paper degradation activity (FPase). Arrhenius plots of cellobiohydrolase were appeared as straight line. The Z-values of cellobiohydrolase calculated by CMCase, FPase and immunoassay were 5.2$^{\circ}C$, 6.4$^{\circ}C$ and 5.8$^{\circ}C$, respectively. The thermodynamic parameters obtained from FPase were better agreement with those of immunoassay than CMCase assay.

• Microbiology and Biotechnology Letters
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• v.16 no.3
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• pp.226-230
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• 1988

Antibody against cellobiohydrolase purified from Trichoderma viride had been obtained by injection to rabbit. The antibody had a high specificity against the cellobiohydroase evidienced by absence of immunological reaction to other isozymes from Trichoderma viride. Assay limit of cellobiohydrolase was 1-10 $\mu\textrm{g}$ by column single immunodiffusion and by enzyme linked immunosorbent assay, it was 10-140 ng and 100-1200 pg when the dilution of antibody was 10$^{-6}$ and 10$^{-5}$, respectively.

• Microbiology and Biotechnology Letters
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• v.19 no.3
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• pp.248-252
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• 1991

- Mutation process and substrate induction of 7i-ichoderma viride cellobiohydrolase were investigated by immunological techniques. Since mutants of Trichodemza uiride such as QM9123, QM9414, TKO41 and MCG77 produced immunologically same cellobiohydrolase, it may be that the mutation be occurred in the reguratory gene rather than in the structural gene of cellobiohydrolase. a-Cellulose and Solka Floc were found to be the best inducer for the production of cellobiohydrolase in Trichodemza viride culture compared to low molecular weight inducer such as carboxylmethyl cellulose and cellobiose.

• Microbiology and Biotechnology Letters
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• v.16 no.3
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• pp.219-225
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• 1988

Two isozymes of cellobiohydrolase and fifteen isozymes of endoglucanase from Trichodema viride QM 9414 were purified by ammonium sulfate fractionation, Sephadex G-100 column chromatography, DEAE-Sephadex A-50 column chromatography and preparative electrophoresis. The purified cellobiohydrolnse had a molecular weight of 71,000 estimated by electrophoresis and amino acid analysis showed its main amino acids to be in the form of aspartic acid and glutamic acid result-ing from its low pI point of 3.81. The optimum pH and temperature were 5.1 and 5$0^{\circ}C$ respectively.

• Korean Journal of Environmental Biology
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• v.17 no.4
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• pp.493-498
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• 1999

To verify the usefulness of enzyme activities as a index for the stability or maturity of organic refuse composting such as grape pomace, Vmax of $\beta$-glucosidase, cellobiohydrolase and alkaline phosphatase were measured. The peak values of all measured enzymes at the initial stage of composting were probably associated with easily degradable organic matter in the grape pomace and decreased gradually. But the activities of $\beta$-glucosidase and cellobiohydrolase were increased again rapidly whereas that of alkaline phosphatase remained approximately constant after 60 composting days. These results suggest that the increase of enzyme activities during the later periods of grape pomace composting process could be used as a index for their stability.

• Applied Biological Chemistry
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• v.28 no.3
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• pp.162-166
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• 1985

Three isozymes of Carboxymethyl Cellulase $(FI^*,\;FII^*,\;FIII)$ and two fractious of ${\beta}-1,4-D-Cellobiohydrolase$(CI, CIl) from Aspergillus niger were purified by Sephadex G-150, DEAE-Sephadex and Sephadex G-75 column chromatography. From the results of enzymatic hydrolysis and X-ray diffraction, ${\beta}-1,4-D-Cellobiohyarolase$ has a high activity toward highly crystalline cellulose such as filter paper and acts synergistically with Cx enzyme.

• Journal of Industrial Technology
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• v.36
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• pp.33-37
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• 2016

Cellulose binding domains (CBDs) of cellulases are thought to assist in the hydrolysis of insoluble crystalline cellulose. To gain sufficient amount of CBDs, the self-cleavable intein tag was used for expression and purification of Trichoderma reesei cellobiohydrolase I CBD in E. coli. Synthetic CBD genes, CBD or linker-CBD were cloned into expression vector pTYB11. Recombinant CBDs were successfully purified by intein mediated purification with an affinity chitin-binding domain. The final yields of recombinant CBD and linker-CBD were 3.2 mg/L and 1.4 mg/L, respectively. The functional bindings of recombinant CBDs were confirmed by Avicel binding experiments. The simple and easy purification method using self-cleavable intein tag can be further used in pretreatment of crystalline cellulose or characterization of engineered CBDs.