Molecular cloning, characterisation and expression of a gene encoding cellobiose dehydrogenase from Termitomyces clypeatus

Banerjee, Sanchita; Roy, Amit

doi:10.1016/j.genrep.2021.101063

Cited by 5 publications

(3 citation statements)

References 44 publications

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“…The molecular weight of CDH is found to be 100 kD. These results are consistent with other reports which have shown CDH to be a monomeric enzyme 6,10,16,19 .…”

Section: Production and Purification Of Cdhsupporting

confidence: 91%

Cellobiose Dehydrogenase from Schizophyllum commune Bcc26414: Purification and Characterization

Yadav,

Nighojkar

2023

Biosci., Biotech. Res. Asia

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CDH, an enzyme produced by wood-decomposing fungi, has diverse applications in biosensor production, bioremediation, and biomedical industries. In this study, CDH from Schizophyllum commune BCC26414 was purified using ammonium sulfate precipitation, DEAE-cellulose chromatography, and Sephadex G-200 chromatography. The purification fold achieved was 65.81 with a specific activity of 1612.34 U/mg. The purity and molecular weight of CDH was confirmed using native and SDS PAGE. Optimal temperature and pH were found to be 30°C and 5, respectively. The purified CDH exhibited stability over a wide pH range (3.5 to 6.5) for 24 hrs and retained complete activity at 40°C, with reduced activity at 50°C when observed for 150 min. KCl, MgSO4, ZnSO4, and NiCl2 at a concentration of 5 mM enhanced CDH activity and HgCl2 and CuSO4 inhibited the enzyme activity. The kinetic constants, Km and Vmax of CDH for lactose were observed to be 125 mM and 13.26 U/ml, respectively. The purified CDH may be utilized commercially in various applications.

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“…The molecular weight of CDH is found to be 100 kD. These results are consistent with other reports which have shown CDH to be a monomeric enzyme 6,10,16,19 .…”

Section: Production and Purification Of Cdhsupporting

confidence: 91%

Cellobiose Dehydrogenase from Schizophyllum commune Bcc26414: Purification and Characterization

Yadav,

Nighojkar

2023

Biosci., Biotech. Res. Asia

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“…In addition, the optimal temperature of gCDH was determined to be 45°C, and the optimal PH was 4.0. CDH from various sources had been successfully expressed in Pichia pastoris , Trichoderma reesei , and A. niger ( Ma et al, 2017 ; Banerjee and Roy, 2021 ). Ma et al (2017) and Bey et al (2011) expanded the scale of fermentation and made the activity of CDH reach a relatively high level.…”

Section: Discussionmentioning

confidence: 99%

Expression of cellobiose dehydrogenase gene in Aspergillus niger C112 and its effect on lignocellulose degrading enzymes

Zhong,

Guo,

et al. 2024

Front. Microbiol.

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Cellobiose dehydrogenase (CDH) is one of the cellulase auxiliary proteins, which is widely used in the field of biomass degradation. However, how to efficiently and cheaply apply it in industrial production still needs further research. Aspergillus niger C112 is a significant producer of cellulase and has a relatively complete lignocellulose degradation system, but its CDH activity was only 3.92 U. To obtain a recombinant strain of A. niger C112 with high cellulases activity, the CDH from the readily available white-rot fungus Grifola frondose had been heterologously expressed in A. niger C112, under the control of the gpdA promoter. After cultivation in the medium with alkali-pretreated poplar fiber as substrate, the enzyme activity of recombinant CDH reached 36.63 U/L. Compared with the original A. niger C112, the recombinant A. niger transformed with Grifola frondosa CDH showed stronger lignocellulase activity, the activities of cellulases, β-1, 4-glucosidase and manganese peroxidase increased by 28.57, 35.07 and 121.69%, respectively. The result showed that the expression of the gcdh gene in A. niger C112 could improve the activity of some lignocellulose degrading enzymes. This work provides a theoretical basis for the further application of gcdh gene in improving biomass conversion efficiency.

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“…Termitomyces clypeatus CDH intracellular expression in S. cerevisiae was achieved by Banerjee et al [41] in a relatively low but detectable degree of the expressed protein, 0.039 U/mg, with cellobiose as an electron donor [41]. Previous research showed that the secretion signal influences the level of cellobiose dehydrogenase expression.…”

Section: Cdhmentioning

confidence: 99%

State of the Art Technologies for High Yield Heterologous Expression and Production of Oxidoreductase Enzymes: Glucose Oxidase, Cellobiose Dehydrogenase, Horseradish Peroxidase, and Laccases in Yeasts P. pastoris and S. cerevisiae

Crnoglavac Popović,

Stanišić,

Prodanović

2024

Fermentation

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Oxidoreductase (OXR) enzymes are in high demand for biocatalytic applications in the food industry and cosmetics (glucose oxidase (GOx) and cellobiose dehydrogenase (CDH)), bioremediations (horseradish peroxidase (HRP) and laccase (LAC)), and medicine for biosensors and miniature biofuel cells (GOx, CDH, LAC, and HRP). They can be used in a soluble form and/or within the yeast cell walls expressed as chimeras on the surface of yeast cells (YSD), such as P. pastoris and S. cerevisiae. However, most of the current studies suffer from either low yield for soluble enzyme expression or low enzyme activity when expressed as chimeric proteins using YSD. This is always the case in studies dealing with the heterologous expression of oxidoreductase enzymes, since there is a requirement not only for multiple OXR gene integrations into the yeast genome (super transformations), and codon optimization, but also very careful design of fermentation media composition and fermentation conditions during expression due to the need for transition metals (copper and iron) and metabolic precursors of FAD and heme. Therefore, scientists are still trying to find the optimal formula using the above-mentioned approaches; most recently, researcher started using protein engineering and directed evolution to increase in the yield of recombinant enzyme production. In this review article, we will cover all the current state-of-the-art technologies and most recent advances in the field that yielded a high expression level for some of these enzymes in specially designed expression/fermentation systems. We will also tackle and discuss new possibilities for further increases in fermentation yield using cutting-edge technologies such as directed evolution, protein and strain engineering, high-throughput screening methods based on in vitro compartmentalization, flow cytometry, and microfluidics.

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Molecular cloning, characterisation and expression of a gene encoding cellobiose dehydrogenase from Termitomyces clypeatus

Cited by 5 publications

References 44 publications

Cellobiose Dehydrogenase from Schizophyllum commune Bcc26414: Purification and Characterization

Cellobiose Dehydrogenase from Schizophyllum commune Bcc26414: Purification and Characterization

Expression of cellobiose dehydrogenase gene in Aspergillus niger C112 and its effect on lignocellulose degrading enzymes

State of the Art Technologies for High Yield Heterologous Expression and Production of Oxidoreductase Enzymes: Glucose Oxidase, Cellobiose Dehydrogenase, Horseradish Peroxidase, and Laccases in Yeasts P. pastoris and S. cerevisiae

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