A diverse set of family 48 bacterial glycoside hydrolase cellulases created by structure‐guided recombination

• DOI: 10.1111/febs.12032
• Publication Date: 2012-10-17T19:47:48Z
• Authors: Matthew A. Smith; Andrea Rentmeister; Christopher D. Snow; Timothy Wu; Mary F. Farrow; Florence Mingardon; Frances H. Arnold
• Keywords: Protein Engineering; Hydrolase

Sequence diversity within a family of functional enzymes provides platform for elucidating structure-function relationships and protein engineering to improve properties important applications. Access nature's vast sequence is often limited by the fact that only few have been characterized in given family. Here, we recombined catalytic domains three glycoside hydrolase 48 bacterial cellulases (Cel48; EC 3.2.1.176) - Clostridium cellulolyticum CelF, stercorarium CelY, thermocellum CelS create diverse library Cel48 with an average 106 mutations from closest native enzyme. Within this set, found large variations such as temperature range, stability, specific activity on crystalline cellulose. We showed status stability were predictable simple linear models sequence-property data: fragments contributed additively these chimera. Using this, correctly predicted sequences stable any described date. The characterization 60 active chimeras expands number 13 73. Our work illustrates role structure-guided recombination can play helping identify sequence-function supplementing natural synthetic diversity.