Development of bi-functional chimeric enzyme (CtGH1-L1-CtGH5-F194A) from endoglucanase (CtGH5) mutant F194A and β-1,4-glucosidase (CtGH1) from Clostridium thermocellum with enhanced activity and structural integrity

Nath, Priyanka; Dhillon, Arun; Kumar, Krishan; Sharma, Kananbala; Jamaldheen, Sumitha Banu; Moholkar, Vijayanand S.

doi:10.1016/j.biortech.2019.03.051

Cited by 34 publications

(10 citation statements)

References 20 publications

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“…Since the last decades, designing and constructing of chimeric enzymes, generally based on the association of two enzymes in a bifunctional protein complex, were widely explored and pursued for different purposes including the research of synergistic or cooperative effects between partners [ 34 , 35 ], the improvement of inherent biochemical properties such as enzymes stability (pH and thermostability) and activity [ 36 , 37 , 38 ], and the expansion of the specificity to additional substrates [ 39 , 40 ]. In these perspectives, many efforts have been made in the development of fusion enzymes by genetic engineering to optimize the degradation of plants cell wall polymers (polysaccharides, lignin, hemicelluloses, etc.)…”

Section: Discussionmentioning

confidence: 99%

Biochemical Characterization of a Bifunctional Enzyme Constructed by the Fusion of a Glucuronan Lyase and a Chitinase from Trichoderma sp.

Baklouti

Delattre

Pierre

et al. 2020

Life

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Bifunctional enzymes created by the fusion of a glucuronan lyase (TrGL) and a chitinase (ThCHIT42) from Trichoderma sp. have been constructed with the aim to validate a proof of concept regarding the potential of the chimera lyase/hydrolase by analyzing the functionality and the efficiency of the chimeric constructions compared to parental enzymes. All the chimeric enzymes, including or nor linker (GGGGS), were shown functional with activities equivalent or higher to native enzymes. The velocity of glucuronan lyase was considerably increased for chimeras, and may involved structural modifications at the active site. The fusion has induced a slightly decrease of the thermostability of glucuronan lyase, without modifying its catalytic activity regarding pH variations ranging from 5 to 8. The biochemical properties of chitinase seemed to be more disparate between the different fusion constructions suggesting an impact of the linkers or structural interactions with the linked glucuronan lyase. The chimeric enzymes displayed a decreased stability to temperature and pH variations, compared to parental one. Overall, TrGL-ThCHIT42 offered the better compromise in terms of biochemical stability and enhanced activity, and could be a promising candidate for further experiments in the field of fungi Cell Wall-Degrading Enzymes (CWDEs).

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Section: Discussionmentioning

confidence: 99%

Biochemical Characterization of a Bifunctional Enzyme Constructed by the Fusion of a Glucuronan Lyase and a Chitinase from Trichoderma sp.

Baklouti

Delattre

Pierre

et al. 2020

Life

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“…GH5, GH26, and GH44 are three families often found fused to additional GH domains. Fusion of endocellulase and exocellulase (or β-glucosidase) motifs may create intramolecular synergy [ 45 • , 46 ], but this is not a general rule: additional CBMs are often required [ 47 ]. Synergy in these multidomain enzymes is a complex property, and the position of CBM relative to GH is critical [ 48 ].…”

Section: Tier 3 Mfcs: Multi-domain Intramolecular Synergymentioning

confidence: 99%

Multifunctional cellulases are potent, versatile tools for a renewable bioeconomy

Glasgow

Meulen

Kuch

et al. 2021

Current Opinion in Biotechnology

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Enzyme performance is critical to the future bioeconomy based on renewable plant materials. Plant biomass can be efficiently hydrolyzed by multifunctional cellulases (MFCs) into sugars suitable for conversion into fuels and chemicals, and MFCs fall into three functional categories. Recent work revealed MFCs with broad substrate specificity, dual exo-activity/endo-activity on cellulose, and intramolecular synergy, among other novel characteristics. Binding modules and accessory catalytic domains amplify MFC and xylanase activity in a wide variety of ways, and processive endoglucanases achieve autosynergy on cellulose. Multidomain MFCs from Caldicellulosiruptor are heat-tolerant, adaptable to variable cellulose crystallinity, and may provide interchangeable scaffolds for recombinant design. Further studies of MFC properties and their reactivity with plant biomass are recommended for increasing biorefinery yields.

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“…The chimera showed enzyme activity of both fused endoglucanase and glucosidase. It was thermally stable and structurally integral . In another experiment, Chahed et al used recombinant plasmid p PICZαA to transform endoglucanase.…”

Section: Use Of Synthetic Biology Tools and Enzyme Immobilizationmentioning

confidence: 99%

Synthetic Biology and Biocomputational Approaches for Improving Microbial Endoglucanases toward Their Innovative Applications

2021

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Microbial endoglucanases belonging to the β-1–4 glycosyl hydrolase family are useful enzymes due to their vast industrial applications in pulp and paper industries and biorefinery. They convert lignocellulosic substrates to soluble sugars and help in the biodegradation process. Various biocomputational tools can be utilized to understand the catalytic activity, reaction kinetics, complexity of active sites, and chemical behavior of enzyme complexes in reactions. This might be helpful in increasing productivity and cost reduction in industries. The present review gives an overview of some interesting aspects of enzyme design, including computational techniques such as molecular dynamics simulation, homology modeling, mutational analysis, etc., toward enhancing the quality of these enzymes. Moreover, the review also covers the aspects of synthetic biology, which could be helpful in faster and reliable development of useful enzymes with desired characteristics and applications. Finally, the review also deciphers the utilization of endoglucanases in biodegradation and emphasizes the use of diversified protein engineering tools and the modification of metabolic pathways for enzyme engineering.

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Development of bi-functional chimeric enzyme (CtGH1-L1-CtGH5-F194A) from endoglucanase (CtGH5) mutant F194A and β-1,4-glucosidase (CtGH1) from Clostridium thermocellum with enhanced activity and structural integrity

Cited by 34 publications

References 20 publications

Biochemical Characterization of a Bifunctional Enzyme Constructed by the Fusion of a Glucuronan Lyase and a Chitinase from Trichoderma sp.

Biochemical Characterization of a Bifunctional Enzyme Constructed by the Fusion of a Glucuronan Lyase and a Chitinase from Trichoderma sp.

Multifunctional cellulases are potent, versatile tools for a renewable bioeconomy

Synthetic Biology and Biocomputational Approaches for Improving Microbial Endoglucanases toward Their Innovative Applications

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