Structure-based engineering of a pectate lyase with improved specific activity for ramie degumming

摘要: Biotechnological applications of microbial pectate lyases (Pels) in plant fiber processing are promising, eco-friendly substitutes for conventional chemical degumming processes. However, to potentiate the enzymes’ use industrial applications, resolving molecular structure elucidate catalytic mechanisms becomes necessary. In this manuscript, we report high resolution (1.45 A) crystal lyase (pelN) from Paenibacillus sp. 0602 apo form. Through sequence alignment and structural superposition with other members polysaccharide (PL) family 1 (PL1), determined that pelN shares characteristic right-handed β-helix is structurally similar PL1 family, while exhibiting key differences terms substrate binding residues. Then, based on information alignments PLs, engineered a novel pelN. Our rational design yielded mutant temperature enzymatic activity optimally shifted 67.5 60 °C. Most importantly, displayed both higher specific ramie ability when compared wild-type enzyme. Altogether, our method shows great potential applications. Moreover, expect reported high-resolution provide solid foundation future rational, structure-based engineering genetically enhanced pelNs.