Molecular Cloning and Sequencing of AlkalophilicCellulosimicrobium cellulans CKMX1 Xylanase Gene Isolated from Mushroom Compost and Characterization of the Gene Product

Walia, Abhishek; Mehta, Preeti; Guleria, Shiwani; Chauhan, Anjali; Shirkot, C. K.

doi:10.1590/s1516-89132015060319

Cited by 9 publications

(4 citation statements)

References 30 publications

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“…Therefore, recombinant DNA techniques must be employed as an excellent tool for the construction of genetically modified strains of microbes with selected characteristics for enzyme production. In this case, isolation and cloning of xylanase gene designate an important step in the engineering of the most efficient microorganism (Walia et al 2015a). Till date, xylanase gene isolated from various microorganisms have been cloned and expressed into suitable hosts with various objectives.…”

Section: Molecular Cloning and Expression Of Xylanase Genementioning

confidence: 99%

“…There are reports related to cloning and expression of xylanase from bacteria such as Cellulosimicrobium sp. (Kim et al 2012), Cellulosimicrobium cellulans (Walia et al 2015a), Nesterenkoniaxinjiangensis (Kui et al 2010), Thermobifida halotolerans (Zhang et al 2012) and Bacillus subtilis into a non-cellulase producing strain of E. coli. The main targets of cloning are the improvement of fermentation processes of industrially important xylose fermenting microbes, by introducing genes for xylanase and xylosidase, for enhancing of xylanolytic activity devoid of cellulase activity.…”

Section: Molecular Cloning and Expression Of Xylanase Genementioning

confidence: 99%

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Microbial xylanases and their industrial application in pulp and paper biobleaching: a review

et al. 2017

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Xylanases are hydrolytic enzymes which cleave the β-1, 4 backbone of the complex plant cell wall polysaccharide xylan. Xylan is the major hemicellulosic constituent found in soft and hard food. It is the next most abundant renewable polysaccharide after cellulose. Xylanases and associated debranching enzymes produced by a variety of microorganisms including bacteria, actinomycetes, yeast and fungi bring hydrolysis of hemicelluloses. Despite thorough knowledge of microbial xylanolytic systems, further studies are required to achieve a complete understanding of the mechanism of xylan degradation by xylanases produced by microorganisms and their promising use in pulp biobleaching. Cellulase-free xylanases are important in pulp biobleaching as alternatives to the use of toxic chlorinated compounds because of the environmental hazards and diseases caused by the release of the adsorbable organic halogens. In this review, we have focused on the studies of structural composition of xylan in plants, their classification, sources of xylanases, extremophilic xylanases, modes of fermentation for the production of xylanases, factors affecting xylanase production, statistical approaches such as Plackett Burman, Response Surface Methodology to enhance xylanase production, purification, characterization, molecular cloning and expression. Besides this, review has focused on the microbial enzyme complex involved in the complete breakdown of xylan and the studies on xylanase regulation and their potential industrial applications with special reference to pulp biobleaching, which is directly related to increasing pulp brightness and reduction in environmental pollution.

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Section: Molecular Cloning and Expression Of Xylanase Genementioning

confidence: 99%

Section: Molecular Cloning and Expression Of Xylanase Genementioning

confidence: 99%

Microbial xylanases and their industrial application in pulp and paper biobleaching: a review

et al. 2017

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“…The xylanase enzymes used in bio-bleaching process should be characterized by active alkaline pH, high temperature, and do not contain a cellulolytic enzyme to avoid cellulose fiber degradation [47]. The researchers were interested in using xylanase enzyme in the bleaching process since it helps to reduce the consumption of chlorine compounds [48,49].…”

Section: Bio-bleaching Of Wastepaper By Xylanase Enzymementioning

confidence: 99%

Xylanase from thermotolerant Bacillus haynesii strain, synthesis, characterization, optimization using Box-Behnken Design, and biobleaching activity

Bakry

Salem

Atta

et al. 2022

Biomass Conv. Bioref.

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The economic value of xylanolytic enzymes is derived from their use in a variety of industrial processes, which necessitates a cost-effective manufacturing procedure. In the current study, forty bacterial isolates were isolated from water samples and investigate their efficacy to producing xylanase enzyme. The most potent bacterial isolate was identified by sequencing and amplifications of 16Sr RNA gene as Bacillus haynesii strain K6. The impacts of various culture conditions on the productivity of xylane were examined. Data showed that the highest xylanase production was achieved at pH 7, in presence of 3 g/L xylan, 5 g/L peptone, and incubated at 40 °C for 24 h. The Box-Behnken model was used to find the best parameters for the relevant variables, and the results revealed an increase in xylanase production with values of 35.02 U/mL. The maximum precipitation of xylanase from the optimized culture was attained by ammonium sulfate (60%) followed by purification using dialysis and sephadex G100 column chromatography. The purified xylanase had a 12-fold enrichment, with a specific activity of 84 U/mg and a molecular weight approximately 439 KDa determined by thin-layer chromatography (TLC)/mass spectrometry. The amino acid analysis of the purified xylanase enzyme revealed the presence of 15 amino acids, with the highest concentrations of 1940 and 1520 mg/L for proline and cysteine, respectively. Finally, the physical properties of wastepaper pulp were improved after treatment with xylanase enzyme. The whiteness and softness of xylanase-treated wastepaper were improved with percentages of 34.6% and 16.2%, respectively. Therefore, we recommend the use of xylanase enzyme in the bleaching process as it is a biologically synthetic material, safe, and suitable for industrial use, and it reduces the use of harmful chemicals.

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“…Developments in biotechnology, such as protein engineering and directed evolution, revolutionized the probability of producing novel enzymes by introducing or modifying the capability of specific genes (Sandgren et al, 2013;Chen et al, 2014;Walia et al, 2015;Enkhbaatar et al, 2016;Castillo et al, 2016). Advances in biotechnology is providing a plethora of enzymes displaying new activities and having adaptability to a range of conditions leading to their increased adoption for industrial purposes (Diaz et al, 2004;Brzezinska et al, 2013;Sriyapai et al, 2013;Munar et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

Purification and molecular characterization of chitinases from soil actinomycetes

Das¹,

Kumar²,

Kumar³

et al. 2017

Afr. J. Microbiol. Res.

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Microbial extracellular chitinases are used in agriculture as effective biocontrol agents and in waste degradation, pharmaceutical and food industry. Actinomycetes are widely tapped group for production of extracellular chitinases. In the present study, approximately 260 actinomycetes were isolated from various ecological habitats was subjected to primary analyses and screened for production of chitinase by plate assay method. Diameter of zones of hydrolysis ranged from 8 to 16 mm. Based on the results, isolates 130, 194, 184, NRRLB 24916 (Streptomyces mexicanus) and NRRLB 16746 (Streptomyces albidoflavus, positive control) were selected for secondary screening and purification. Enzyme activity was estimated in crude cell free extract and partially purified samples. Activity ranged from 7.16 to 14.12 IU/ml (in crude extracts) and 12.1 to 23.10 IU/ml (in partially purified samples). In case of highest chitinase producing isolate 130, effect of various fermentation conditions (pH, temperature and substrate concentration) was studied in crude extract. Furthermore, complete purification of isolate 130 was done by column chromatography and the activity in purified fraction was found to be 32.12 IU/ml. The K m and V max values of the purified fractions for isolate 130 were 2.11 µg/ml/min and 53.11mg/ml respectively. This shows that the enzyme has high affinity for the substrate. SDS gel electrophoresis of the purified fraction showed presence of single band of approximately 65 to 70 kDa. Analyses of purified chitinase were done using MS/MS technique. N-terminal sequence corresponded to chitinase, the gene encodes a protein of 453 amino acid residues. Comparison of deduced amino acid sequence to other chitinases in the database indicated that enzyme showed 70% similarity with chitinase from Streptomyces plicatus and belongs to glycoside hydrolase family 18. Homology modeling showed that the enzyme was folded into a domain of (α/β) 8 barrel structure. Identification of secondary structure was done by CD spectroscopy. Isolate 130 was capable of degrading biodegradable wastes such as crustacean shells.

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Molecular Cloning and Sequencing of AlkalophilicCellulosimicrobium cellulans CKMX1 Xylanase Gene Isolated from Mushroom Compost and Characterization of the Gene Product

Abstract: A xylanolytic bacterium was isolated from mushroom compost by using enrichment

Cited by 9 publications

References 30 publications

Microbial xylanases and their industrial application in pulp and paper biobleaching: a review

Microbial xylanases and their industrial application in pulp and paper biobleaching: a review

Xylanase from thermotolerant Bacillus haynesii strain, synthesis, characterization, optimization using Box-Behnken Design, and biobleaching activity

Purification and molecular characterization of chitinases from soil actinomycetes

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