Structural and functional domains of cellobiohydrolase I from trichoderma reesei

Abuja, Peter M.; Schmuck, Maria; Pilz, Ingrid; Tomme, Peter; Claeyssens, Marc; Esterbauer, Hermann

doi:10.1007/bf00254721

Cited by 131 publications

(70 citation statements)

References 21 publications

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“…10 In the cellulose degradation system of the ascomycetes fungus Trichoderma reesei, a dominant component of the secreted proteins is glycoside hydrolase (GH) family 7 cellobiohydrolase (TrCel7A, formerly known as CBH I), which produces cellobiose from the reducing end of cellulose. 11−13 TrCel7A has a two-domain structure, 14 with a catalytic domain (CD) and a cellulose-binding domain (CBD) belonging to the (A) Phylogenetic tree of the sequence-based alignment of Cel7s classified as "characterized" by CAZY (http://www.cazy.org/). Sequences were aligned and the tree was made with ClustalW2 server default settings.…”

Section: ■ Introductionmentioning

confidence: 99%

Trade-off between Processivity and Hydrolytic Velocity of Cellobiohydrolases at the Surface of Crystalline Cellulose

et al. 2014

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Analysis of heterogeneous catalysis at an interface is difficult because of the variety of reaction sites and the difficulty of observing the reaction. Enzymatic hydrolysis of cellulose by cellulases is a typical heterogeneous reaction at a solid/liquid interface, and a key parameter of such reactions on polymeric substrates is the processivity, i.e., the number of catalytic cycles that can occur without detachment of the enzyme from the substrate. In this study, we evaluated the reactions of three closely related glycoside hydrolase family 7 cellobiohydrolases from filamentous fungi at the molecular level by means of high-speed atomic force microscopy to investigate the structure−function relationship of the cellobiohydrolases on crystalline cellulose. We found that high moving velocity of enzyme molecules on the surface is associated with a high dissociation rate constant from the substrate, which means weak interaction between enzyme and substrate. Moreover, higher values of processivity were associated with more loop regions covering the subsite cleft, which may imply higher binding affinity. Loop regions covering the subsites result in stronger interaction, which decreases the velocity but increases the processivity. These results indicate that there is a trade-off between processivity and hydrolytic velocity among processive cellulases.

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Section: ■ Introductionmentioning

confidence: 99%

Trade-off between Processivity and Hydrolytic Velocity of Cellobiohydrolases at the Surface of Crystalline Cellulose

et al. 2014

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“…Most fungal cellulases have a common structural organization where the main part of the enzyme, the catalytic core, is connected through a heavily glycosylated linker region to a small cellulose-binding domain (CBD) [4]. This gives the whole molecule an elongated tadpole shape (180 A in the case of CBH 1) [5]. The three-dimensional *Corresponding author.…”

Section: Introductionmentioning

confidence: 99%

“…Abbreviations: EG 1, endoglucanase 1; CBH 1, cellobiohydrolase 1; pNPL, p-nitro-phenyl-~-D-lactoside; CBD, cellulose-binding domain; CSP, chiral stationary phase structure of the binding domain has been solved in only one case, but there are good reasons to believe that the others are all folded in the same way on the basis of sequence identity [2,5]. The crystal structure of the CBH 1 catalytic domain reveals a large domain (434 residues) with the dimensions 60 × 50 × 40 A.…”

Section: Introductionmentioning

confidence: 99%

The active sites of cellulases are involved in chiral recognition: a comparison of cellobiohydrolase 1 and endoglucanase 1

et al. 1996

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The cellulases ceHobiohydrolase 1 (CBH 1) and endoglucanase 1 (EG 1) from the fungus Trichoderma reesei are closely related with 40% sequence identity and very similar in structure. In CBH 1 the active site is enclosed by long loops and some antiparallel ~strands forming a 40 A long tunnel, whereas in EG 1 part of those loops are missing so that the enzyme has a more common active site groove. Both enzymes were immobilized on silica and these materials were used as chiral stationary phases for chromatographic separation of the enantiomers of two chiral drugs, propranolol and alprenolol. The CBH 1 phase showed much better resolution than did the EG 1 phase, suggesting that the tunnel structure of the protein may play an important role in the chiral separation. The chiral compounds were found to be competitive inhibitors of both enzymes when p-nitrophenyl lactoside (pNPL) was used as substrate. (S)-enantiomers showed stronger inhibitory effects and also longer retention time on the stationary phases than the (R)-enantiomers. The consistency between kinetic data and retention on the stationary phases clearly shows that the enzymatically active sites of CBH 1 and EG 1 are involved in chiral recognition.

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“…We expect this type of surface to be relevant to the cellulose/cellulase system. The shape of the cellulases in the model was defined according to small-angle X-ray-scattering (SAXS) imaging [20,21] (Fig. 1).…”

Section: Methods Preparation Of Bacterial Microcrystalline Cellulosementioning

confidence: 99%

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Nutt

Sild²,

Pettersson³

et al. 1998

European Journal of Biochemistry

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The chain-end preference and processivity of the cellulases 1,4-β-D-glucan-cellobiohydrolase I (CBH I) and 1,4-β-D-glucan-cellobiohydrolase II (CBH II) from Trichoderma reesei and 1,4-β-D-glucancellobiohydrolase 50 (CBH 50) and 1,4-β-D-glucan-cellobiohydrolase 58 (CBH 58) from Phanerochaete chrysosporium were studied by comparing experimental degradation data on reducing end-labelled bacterial microcrystalline cellulose with computer simulations of different models. Our results with T. reesei and P. chrysosporium cellulases show that there is a common pattern of hydrolysis for CBH-I-type enzymes and another clearly distinguishable pattern for CBH-II-type enzymes, especially in the initial part of the progress curve.Keywords : cellulase ; cellulose; hydrolysis ; simulation ; mode of action.In nature, the breakdown of cellulose is carried out mainly by fungi and bacteria. These organisms degrade crystalline cellulose by means of several enzymes acting at the ends (exocellulases) or randomly (endoglucanases) on the cellulose chains. Cellulases, like many other enzymes attacking insoluble or 'larger-than-enzyme' polymeric substrates, have a characteristic two-domain organisation : a catalytic domain connected by a flexible linker to a separate cellulose-binding domain [1]. Effective degradation of native cellulose requires cooperation between different functional types of cellulases and displays two distinctive modes of synergism; namely, cooperation among endoglucanases and cellobiohydrolases (CBHs) (endo/exo synergism), where the endoglucanases are supposed to create a large number of free ends susceptible to exoglucanase action [2Ϫ4], and cooperation among different CBHs [5Ϫ7]. The molecular basis for this latter synergism is not yet understood, mainly because the modes of action of the individual enzymes on crystalline cellulose are not known. By definition, CBHs attack the cellulose chain from the non-reducing end [8]. Recent data indicate, however, that the exhaustively studied Trichoderma reesei 1,4-β-D-glucan-cellobiohydrolase I (CBH I) preferentially attacks the reducing end of cellooligosaccharides, whereas the similarly well-characterised 1,4-β-D-glucan-cellobiohydrolase II (CBH II) from the same organism indeed prefers the non-reducing end [9,10].

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Structural and functional domains of cellobiohydrolase I from trichoderma reesei

Cited by 131 publications

References 21 publications

Trade-off between Processivity and Hydrolytic Velocity of Cellobiohydrolases at the Surface of Crystalline Cellulose

Trade-off between Processivity and Hydrolytic Velocity of Cellobiohydrolases at the Surface of Crystalline Cellulose

The active sites of cellulases are involved in chiral recognition: a comparison of cellobiohydrolase 1 and endoglucanase 1

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