Determination of bioethanol production potential from lignocellulosic biomass using novel Cel-5m isolated from cow rumen metagenome.

We are reporting the novel cellulase named Cel-5M from rumen metagenome. The deduced amino acid sequence and biochemical characterization suggested that Cel-5M is endoglucanase from the GH5 family with multifunctional potential. Cel-5M showed similarity to non-characterized proteins and the genus Prevotella as parental organism. The 957 bp ORF encoding Cel-5M was cloned and overexpressed in E. coli and purified. The recombinant Cel-5M showed maximum activity at pH 6.0 and 40 °C. It retained more than 80% activity between 4 and 7 pH and 65% thermostability between 30 and 70 °C. Cel-5M showed activity on various substrates like CMC, Filter paper, Avicel, Xylan, β-Mannan, and Glucopyranoside, which confirmed its multifunctional characteristics and classifies as member of subfamily 4 of GH5 (GH5_4). LCB hydrolysis potential of Cel-5M was studied using wheat straw (10% w/v). Alkali-treated and steam-exploded wheat straws were inoculated with 1 mg/g Cel-5M, 2% yeast in a reaction mixture and SSF at 10% w/v loading rate. The ethanol yield 0.46 g/g and 0.43 g/g of cellulose obtained after 72 h fermentation in alkali-treated and steam-exploded wheat straw, respectively. Cel-5M is novel multifunction cellulase belongs GH-5 endoglucanase from rumen origin can be employed for bioethanol based biofuel production.

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