A novel endo-glucanase from the thermophilic bacterium Geobacillus sp. 70PC53 with high activity and stability over a broad range of temperatures

Abstract

A thermophilic Geobacillus bacterium secreting high activity of endo-glucanase (EC 3.2.1.4) was isolated from rice straw compost supplemented with pig manure. A full-length gene of 1,104 bp, celA, encoding this glycosyl hydrolase family 5 endo-glucanase of 368 amino acids was isolated. No related gene from Geobacillus has been reported previously. The recombinant CelA expressed in Escherichia coli had an optimal activity at 65°C and pH 5.0, and it exhibited tenfold greater specific activity than the commercially available Trichoderma reesei endo-glucanase. CelA displayed activity over a broad temperature range from 45 to 75°C and was a thermostable enzyme with 90% activity retained after heating at 65°C for 6 h. Interestingly, CelA activity could be enhanced by 100% in the presence of 2 mM MnSO₄. CelA had high specific activity over β-d-glucan from barley and Lichenan, making it a potentially useful enzyme in biofuel and food industries.