Expression of eg9 gene encoding endoglucanase GH8 derived from metagenomic DNA data of bacteria in humus around white rot fungi degrading woods, in Escherichia coli

Endoglucanase is an important enzyme participating in the hydrolysis of cellulose into oligosaccharides or glucoses can be used a variety industrial fields. A gene (code GL0183420 designated as eg9) 1398 bp encoding for endoglucanase family GH8 was identified from metagenomic DNA data bacteria humus surrounding wood hydrolyzed by white rot fungi. The 126 terminal nucleotides at 5' encode signal peptide gram-negative and next 1269 GH8. In this study, eg9 (lack sequence peptid) coding mature codon optimized expression E. coli, artificical synthesized, then inserted pET22b(+) coli strains. Through survey 5 expressive strains, EG9 overexpressed Rosetta C43 However, 30oC LB medium, most were expressed insoluble fraction, which best strain synthesis EG9. large range OD600 0.2 to 1. After reducing culture temperature 25°C, 50% soluble fraction exhibited good activity CMC substrate on agar plate. investigation media showed that PE suitable medium recombinant grow synthesize well various IPTG concentrations 0.05 0.9 mM. These results offer great potential production properties research.