Cellobiose dehydrogenase of <i>Chaetomium</i> sp. INBI 2‐26(–): Structural basis of enhanced activity toward glucose at neutral pH

Vasil'chenko, L. G.; Karapetyan, K.N.; Yershevich, Olga P.; Ludwig, Roland; Zámocký, Marcel; Peterbauer, Clemens K.; Haltrich, Dietmar; Rabinovich, M. A.

doi:10.1002/biot.201000373

Cited by 6 publications

(3 citation statements)

References 42 publications

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“…The rather strict selectivity of the originally investigated class I basidiomycete CDHs for cellodextrins and lactose [60, 84] mainly found application in lactose biosensors for the dairy industry [55, 58, 85] or cellobiose biosensors that can be used to follow cellulose hydrolysis caused by cellulose-hydrolysing enzymes [54, 86]. However, when it was realised that the recently discovered ascomycete CDHs (class IIA and IIB) could also have high turnover rates for glucose (and a series of other mono- and oligosaccharides) and especially at neutral pH values (in contrast to class I CDHs that work best under slightly acidic conditions) an increased interest in CDH for application in biosensors and biofuel cells appeared [30, 42, 44, 49–51, 56, 57, 87–92]. …”

Section: Direct Electron Transfermentioning

confidence: 99%

Cellobiose dehydrogenase modified electrodes: advances by materials science and biochemical engineering

et al. 2013

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The flavocytochrome cellobiose dehydrogenase (CDH) is a versatile biorecognition element capable of detecting carbohydrates as well as quinones and catecholamines. In addition, it can be used as an anode biocatalyst for enzymatic biofuel cells to power miniaturised sensor–transmitter systems. Various electrode materials and designs have been tested in the past decade to utilize and enhance the direct electron transfer (DET) from the enzyme to the electrode. Additionally, mediated electron transfer (MET) approaches via soluble redox mediators and redox polymers have been pursued. Biosensors for cellobiose, lactose and glucose determination are based on CDH from different fungal producers, which show differences with respect to substrate specificity, pH optima, DET efficiency and surface binding affinity. Biosensors for the detection of quinones and catecholamines can use carbohydrates for analyte regeneration and signal amplification. This review discusses different approaches to enhance the sensitivity and selectivity of CDH-based biosensors, which focus on (1) more efficient DET on chemically modified or nanostructured electrodes, (2) the synthesis of custom-made redox polymers for higher MET currents and (3) the engineering of enzymes and reaction pathways. Combination of these strategies will enable the design of sensitive and selective CDH-based biosensors with reduced electrode size for the detection of analytes in continuous on-site and point-of-care applications.

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Section: Direct Electron Transfermentioning

confidence: 99%

Cellobiose dehydrogenase modified electrodes: advances by materials science and biochemical engineering

et al. 2013

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“…The same optimums as for cellobiose were obtained by both enzymes with glucose as substrate, although the activity of the basidiomycetous Sr CDH in this reaction was two orders of magnitude lower than that of neutral ascomycetous Ch CDH (cf. [38]).…”

Section: Resultsmentioning

confidence: 99%

High‐throughput screening for cellobiose dehydrogenases by Prussian Blue in situ formation

Vasil'chenko

Ludwig

Yershevich

et al. 2012

Biotechnology Journal

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Extracellular fungal flavocytochrome cellobiose dehydrogenase (CDH) is a promising enzyme for both bioelectronics and lignocellulose bioconversion. A selective high-throughput screening assay for CDH in the presence of various fungal oxidoreductases was developed. It is based on Prussian Blue (PB) in situ formation in the presence of cellobiose (<0.25 mM), ferric acetate, and ferricyanide. CDH induces PB formation via both reduction of ferricyanide to ferrocyanide reacting with an excess of Fe³⁺ (pathway 1) and reduction of ferric ions to Fe²⁺ reacting with the excess of ferricyanide (pathway 2). Basidiomycetous and ascomycetous CDH formed PB optimally at pH 3.5 and 4.5, respectively. In contrast to the holoenzyme CDH, its FAD-containing dehydrogenase domain lacking the cytochrome domain formed PB only via pathway 1 and was less active than the parent enzyme. The assay can be applied on active growing cultures on agar plates or on fungal culture supernatants in 96-well plates under aerobic conditions. Neither other carbohydrate oxidoreductases (pyranose dehydrogenase, FAD-dependent glucose dehydrogenase, glucose oxidase) nor laccase interfered with CDH activity in this assay. Applicability of the developed assay for the selection of new ascomycetous CDH producers as well as possibility of the controlled synthesis of new PB nanocomposites by CDH are discussed.

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“…Homogeneous flavohemoprotein cellobiose dehydrogenase (CDH) from Chaetomium sp . INBI 2‐26 was used for the assay of free cellooligosaccharides accumulation in the course of hydrolysis of fluorogenic substrates with HjCel7A.…”

Section: Methodsmentioning

confidence: 99%

Predominant Nonproductive Substrate Binding by Fungal Cellobiohydrolase I and Implications for Activity Improvement

et al. 2018

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Enzymatic conversion of the most abundant renewable source of organic compounds, cellulose to fermentable sugars is attractive for production of green fuels and chemicals. The major component of industrial enzyme systems, cellobiohydrolase I from Hypocrea jecorina (Trichoderma reesei) (HjCel7A) processively splits disaccharide units from the reducing ends of tightly packed cellulose chains. HjCel7A consists of a catalytic domain (CD) and a carbohydrate-binding module (CBM) separated by a linker peptide. A tunnel-shaped substrate-binding site in the CD includes nine subsites for β-d-glucose units, seven of which (-7 to -1) precede the catalytic center. Low catalytic activity of Cel7A is the bottleneck and the primary target for improvement. Here it is shown for the first time that, in spite of much lower apparent k of HjCel7A at the hydrolysis of β-1,4-glucosidic linkages in the fluorogenic cellotetra- and -pentaose compared to the structurally related endoglucanase I (HjCel7B), the specificity constants (catalytic efficiency) k /K for both enzymes are almost equal in these reactions. The observed activity difference appears from strong nonproductive substrate binding by HjCel7A, particularly significant for MU-β-cellotetraose (MUG ). Interaction of substrates with the subsites -6 and -5 proximal to the nonconserved Gln101 residue in HjCel7A decreases K by >1500 times. HjCel7A can be nonproductively bound onto cellulose surface with K ≈2-9 nM via CBM and CD that captures six terminal glucose units of cellulose chain. Decomposition of this nonproductive complex can determine the rate of cellulose conversion. MUG is a promising substrate to select active cellobiohydrolase I variants with reduced nonproductive substrate binding.

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Cellobiose dehydrogenase of Chaetomium sp. INBI 2‐26(–): Structural basis of enhanced activity toward glucose at neutral pH

Cited by 6 publications

References 42 publications

Cellobiose dehydrogenase modified electrodes: advances by materials science and biochemical engineering

Cellobiose dehydrogenase modified electrodes: advances by materials science and biochemical engineering

High‐throughput screening for cellobiose dehydrogenases by Prussian Blue in situ formation

Predominant Nonproductive Substrate Binding by Fungal Cellobiohydrolase I and Implications for Activity Improvement

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