Recombinant lactase with a cellulose binding domain permits facile immobilization onto cellulose with retained activity

Abstract

Because of their potential impact in food and bioprocessing, diagnostics, green(er) chemistry, and waste remediation, new enzyme immobilization technologies continue to be explored. In this work, a recombinant lactase (β-galactosidase) (LacZ) presenting a carbohydrate binding module (LacZ-CBM), was engineered, expressed, and immobilized onto cellulose (LacZ-CBM:cell). Binding density, activity, and pH and temperature activity profiles were characterized in comparison to wild type lactase (LacZ). LacZ-CBM effectively self-immobilized onto cellulose (Sigmacell®) in less than 2 h, demonstrating that recombinant enzymes with cellulose binding modules can enable immobilization onto solid supports without the need for chemical crosslinking agents. The immobilized recombinant LacZ-CBM:cell retained over 30% of its initial activity over 9 cycles. Both immobilized and non-immobilized LacZ-CBM presented similar optimum reaction conditions as wild type lactase, which demonstrates that the addition of a carbohydrate binding module (CBM) to lactase does not alter its optimum reaction conditions. This work has direct relevance to immobilized lactase applications (e.g. lactose reduced milk; oligosaccharide production) and serves as a model for other greener enzyme immobilization applications.