Introduction to Extremozymes

Krishnaraj, R. Navanietha; Sani, Rajesh K.

doi:10.1007/978-3-319-54684-1_1

Cited by 7 publications

(4 citation statements)

References 4 publications

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“…These binding energy values can be used as the standard for predicting the catalytic rates of these enzymes with different lignocellulosic substrates such as corn stover, sugarcane bagasse, switchgrass, wheat straw, etc. Previously, we have reported computational investigations for the screening of the pesticides, drugs, toxicity of nanomaterials and enzymes for bioenergy applications [28][29][30][31][32][33][34]. This strategy will also serve as a tool for screening the other cellulolytic enzymes.…”

Section: Residues In the Allowed Region (%)mentioning

confidence: 99%

Bioprospecting of Thermostable Cellulolytic Enzymes through Modeling and Virtual Screening Method

Krishnaraj¹,

Samanta²,

Kumar³

et al. 2017

Can J Biotech

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Cellulolytic enzymes are promising candidates for the use of cellulose in any bioprocess operations and for the disposal of the cellulosic wastes in an environmentally benign manner. Cellulases from thermophiles have the advantage of hydrolyzing cellulose at wider range of operating conditions unlike the normal enzymes. Herein we report the modeled structures of cellulolytic enzymes (endoglucanase, cellobiohydrolase and β-glucosidase) from a thermophilic bacterium, Clostridium thermocellum and their validation using Root Mean Square Deviation (RMSD) and Ramachandran plot analyses. Further, the molecular interactions of the modeled enzyme with cellulose were analyzed using molecular docking technique. The results of molecular docking showed that the endoglucanase, cellobiohydrolase and β-glucosidase had the binding affinities of -10.7, -9.0 and -10.8 kcal/mol, respectively. A correlation between the binding affinity of the endoglucanase with cellulose and the enzyme activity was also demonstrated. The results showed that the binding affinities of cellulases with cellulose could be used as a tool to assess the hydrolytic activity of cellulases. The results obtained could be used in virtual screening of cellulolytic enzymes based on the molecular interactions with the substrate, and aid in developing systems biology models of thermophiles for industrial biotechnology applications.

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Section: Residues In the Allowed Region (%)mentioning

confidence: 99%

Bioprospecting of Thermostable Cellulolytic Enzymes through Modeling and Virtual Screening Method

Krishnaraj¹,

Samanta²,

Kumar³

et al. 2017

Can J Biotech

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“…Bio-corrosion refers to the deterioration of metal or nonmetal materials caused by the influence of microorganisms typically growing as a biofilm ( 1 ). It is observed in many environments and media including seawater, freshwater, soils, foodstuffs, demineralized water, sewage, aircraft petrol, human plasma and process chemicals ( 2 ). These microbes can be studied to determine how to best protect against these destructive effects and can also provide a rich source of prospected enzymes for the degradation of materials such as plastics.…”

Section: Introductionmentioning

confidence: 99%

Screening putative polyester polyurethane degrading enzymes with semi-automated cell-free expression and nitrophenyl probes

Ahsan,

Wagner,

Varaljay

et al. 2024

Synthetic Biology

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Cell-free expression (CFE) has shown recent utility in prototyping enzymes for discovery efforts. In this work, CFE is demonstrated as an effective tool to screen putative polyester polyurethane degrading enzyme sequences sourced from metagenomic analysis of biofilms prospected on aircraft and vehicles. An automated fluid handler with a controlled temperature block is used to assemble the numerous 30 µL CFE reactions to provide more consistent results over human assembly. In sum, 13 putative hydrolase enzymes from the biofilm organisms as well as a previously verified, polyester-degrading cutinase were expressed using in-house E. coli extract and minimal linear templates. The enzymes were then tested for esterase activity directly in extract using nitrophenyl conjugated substrates, showing highest sensitivity to shorter substrates (4-nitrophenyl hexanoate and 4-nNitrophenyl valerate). This screen identified 10 enzymes with statistically significant activities against these substrates; however, all were lower in measured relative activity, on a CFE volume basis, to the established cutinase control. This approach portends the use of CFE and reporter probes to rapidly prototype, screen and design for synthetic polymer degrading enzymes from environmental consortia. Graphical Abstract

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“…Extremophiles are a type of microorganisms that can grow and reproduce normally under extreme environmental conditions such as extreme temperature, pH, ion concentration, and radioactivity, and they have evolved promising biological components and metabolic pathways to adapt to these complex environments 1 , 2 . In recent years, extremophiles have aroused great interest among researchers due to their ability to catalyze chemical reactions under harsh conditions and other industrial application potentials 3 , 4 , 5 .…”

Section: Introductionmentioning

confidence: 99%

High‐efficiency genome editing of an extreme thermophile Thermus thermophilus using endogenous type I and type III CRISPR‐Cas systems

Wang

Wei

et al. 2022

mLife

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Thermus thermophilus is an attractive species in the bioindustry due to its valuable natural products, abundant thermophilic enzymes, and promising fermentation capacities. However, efficient and versatile genome editing tools are not available for this species. In this study, we developed an efficient genome editing tool for T. thermophilus HB27 based on its endogenous type I-B, I-C, and III-A/B CRISPR-Cas systems. First, we systematically characterized the DNA interference capabilities of the different types of the native CRISPR-Cas systems in T. thermophilus HB27. We found that genomic manipulations such as gene deletion, mutation, and in situ tagging could be easily implemented by a series of genome-editing plasmids carrying an artificial self-targeting mini-CRISPR and a donor DNA responsible for the recombinant recovery. We also compared the genome editing efficiency of different CRISPR-Cas systems and the editing plasmids with donor DNAs of different lengths. Additionally, we developed a reporter gene system for T. thermophilus based on a heat-stable β-galactosidase gene TTP0042, and constructed an engineered strain with a high production capacity of superoxide dismutases by genome modification.

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Introduction to Extremozymes

Cited by 7 publications

References 4 publications

Bioprospecting of Thermostable Cellulolytic Enzymes through Modeling and Virtual Screening Method

Bioprospecting of Thermostable Cellulolytic Enzymes through Modeling and Virtual Screening Method

Screening putative polyester polyurethane degrading enzymes with semi-automated cell-free expression and nitrophenyl probes

High‐efficiency genome editing of an extreme thermophile Thermus thermophilus using endogenous type I and type III CRISPR‐Cas systems

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