Enhanced hydrolytic efficiency of an engineered CBM11-glucanase enzyme chimera against barley β-d-glucan extracts

Abstract

The β-d-glucans are abundant cell wall polysaccharides in many cereals and contain both (1,3)- and (1,4)-bonds. The β-1,3-1,4-glucanases (EC 3.2.1.73) hydrolyze β-(1,4)-d-glucosidic linkages in glucans, and have applications in both animal and human food industries. A chimera between the family 11 carbohydrate-binding module from Ruminoclostridium (Clostridium)thermocellumcelH (RtCBM11), with the β-1,3-1,4-glucanase from Bacillus subtilis (BglS) was constructed by end-to-end fusion (RtCBM11-BglS) to evaluate the effects on the catalytic function and its application in barley β-glucan degradation for the brewing industry. The parental and chimeric BglS presented the same optimum pH (6.0) and temperature (50 °C) for maximum activity. The RtCBM11-BglS showed increased thermal stability and 30% higher hydrolytic efficiency against purified barley β-glucan, and the rate of hydrolysis of β-1,3-1,4-glucan in crude barley extracts was significantly increased. The enhanced catalytic performance of the RtCBM11-BglS may be useful for the treatment of crude barley extracts in the brewing industry.

Keywords: Barley β-glucan degradation; Brewing; Chimeric protein; Family 11 carbohydrate-binding module; GH16 β-1,3-1,4-Glucanase.