Identification and Characterization of a Cellulase-Encoding Gene from the Buffalo Rumen Metagenomic Library

Nguyen, Nhung H. A.; Maruset, Lalita; Uengwetwanit, Tanaporn; Mhuantong, Wuttichai; Harnpicharnchai, Piyanun; Champreda, Verawat; Tanapongpipat, Sutipa; Jirajaroenrat, Kanya; Rakshit, Sudip Kumar; Eurwilaichitr, Lily; Pongpattanakitshote, Somchai

doi:10.1271/bbb.110786

Cited by 48 publications

(44 citation statements)

References 41 publications

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“…In addition, the enzyme retains 70% of the activity in pH values of 8 and 9. Usually cellulases belonging to GH5 family display an optimum for activity at slightly acid pH as observed for the hyperthermophilic CelB from Caldicellulosiruptor saccharolyticus (pH 5.5) [21], Cel5G derived from a soil metagenomic library (pH 4.8) [22], BsCel5A from Bacillus subtilis (pH 6) [23] and cellulase 5 from the buffalo rumen metagenomic library (pH 5.5) [24]. However, this novel cellulase exhibited a bell-shaped curve for pH dependence, shifted towards basic conditions, which can be attractive for biotechnological applications in which conventional acidic cellulases are not suitable.…”

Section: Resultsmentioning

confidence: 99%

Structure and Function of a Novel Cellulase 5 from Sugarcane Soil Metagenome

et al. 2013

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Cellulases play a key role in enzymatic routes for degradation of plant cell-wall polysaccharides into simple and economically-relevant sugars. However, their low performance on complex substrates and reduced stability under industrial conditions remain the main obstacle for the large-scale production of cellulose-derived products and biofuels. Thus, in this study a novel cellulase with unusual catalytic properties from sugarcane soil metagenome (CelE1) was isolated and characterized. The polypeptide deduced from the celE1 gene encodes a unique glycoside hydrolase domain belonging to GH5 family. The recombinant enzyme was active on both carboxymethyl cellulose and β-glucan with an endo-acting mode according to capillary electrophoretic analysis of cleavage products. CelE1 showed optimum hydrolytic activity at pH 7.0 and 50 °C with remarkable activity at alkaline conditions that is attractive for industrial applications in which conventional acidic cellulases are not suitable. Moreover, its three-dimensional structure was determined at 1.8 Å resolution that allowed the identification of an insertion of eight residues in the β8-α8 loop of the catalytic domain of CelE1, which is not conserved in its psychrophilic orthologs. This 8-residue-long segment is a prominent and distinguishing feature of thermotolerant cellulases 5 suggesting that it might be involved with thermal stability. Based on its unconventional characteristics, CelE1 could be potentially employed in biotechnological processes that require thermotolerant and alkaline cellulases.

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Section: Resultsmentioning

confidence: 99%

Structure and Function of a Novel Cellulase 5 from Sugarcane Soil Metagenome

et al. 2013

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“…17) Among NGS technologies, pyrosequencing technology has been instrumental in metagenomic analysis of the microbial communities from environmental samples. 14,[18][19][20][21] In line with this, many studies regarding the microbial community analysis of traditional Korean foods also implemented pyrosequencing technologies. 3,5,6,12,13,15,22,23) In the present study, we executed pyrosequencingbased metagenomic analysis of the bacterial community resident in gajami-sikhae.…”

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confidence: 99%

Pyrosequencing analysis of microbiota reveals that lactic acid bacteria are dominant in Korean flat fish fermented food,gajami-sikhae

Kim

Turner

et al. 2014

Bioscience, Biotechnology, and Biochemistry

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“…To ensure identification of different hemicellulases, more than 50 000 colonies per library were assayed. We found 22 clones with endoxylanase activity and 125 with β‐xylosidase activity (Table ); this indicated that the rate of recovery of hemicellulases was in the range of 1 in 0.5 Mb, much higher than in other reports in which hemicellulases hits were in the range of 1 in 8 Mb or more (Gao et al ., ; Nguyen et al ., ). We consider that our high rate of success was due to the specific enrichment of rumen microbes that used industrial substrates rich in hemicellulose as a carbon and energy source.…”

Section: Resultsmentioning

confidence: 97%

Ruminal metagenomic libraries as a source of relevant hemicellulolytic enzymes for biofuel production

Duque

Daddaoua

Cordero

et al. 2018

Microbial Biotechnology

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SummaryThe success of second‐generation (2G) ethanol technology relies on the efficient transformation of hemicellulose into monosaccharides and, particularly, on the full conversion of xylans into xylose for over 18% of fermentable sugars. We sought new hemicellulases using ruminal liquid, after enrichment of microbes with industrial lignocellulosic substrates and preparation of metagenomic libraries. Among 150 000 fosmid clones tested, we identified 22 clones with endoxylanase activity and 125 with β‐xylosidase activity. These positive clones were sequenced en masse, and the analysis revealed open reading frames with a low degree of similarity with known glycosyl hydrolases families. Among them, we searched for enzymes that were thermostable (activity at > 50°C) and that operate at high rate at pH around 5. Upon a wide series of assays, the clones exhibiting the highest endoxylanase and β‐xylosidase activities were identified. The fosmids were sequenced, and the corresponding genes cloned, expressed and proteins purified. We found that the activity of the most active β‐xylosidase was at least 10‐fold higher than that in commercial enzymatic fungal cocktails. Endoxylanase activity was in the range of fungal enzymes. Fungal enzymatic cocktails supplemented with the bacterial hemicellulases exhibited enhanced release of sugars from pretreated sugar cane straw, a relevant agricultural residue.

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Identification and Characterization of a Cellulase-Encoding Gene from the Buffalo Rumen Metagenomic Library

Cited by 48 publications

References 41 publications

Structure and Function of a Novel Cellulase 5 from Sugarcane Soil Metagenome

Structure and Function of a Novel Cellulase 5 from Sugarcane Soil Metagenome

Pyrosequencing analysis of microbiota reveals that lactic acid bacteria are dominant in Korean flat fish fermented food,gajami-sikhae

Ruminal metagenomic libraries as a source of relevant hemicellulolytic enzymes for biofuel production

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