University of Piemonte Orientale, Dipartimento di Scienze e Innovazione Tecnologica Complesso Universitario S. Giuseppe, Piazza S. Eusebio 5, Vercelli, 13100, Italy.
University of Piemonte Orientale, Dipartimento di Scienze e Innovazione Tecnologica Viale T. Michel 11, Alessandria, 15121, Italy.
Sci Rep. 2018 Mar 13;8(1):4402. doi: 10.1038/s41598-018-22769-w.
Lignocellulosic biomass (LCB) is a low-cost and abundant source of fermentable sugars. Enzymatic hydrolysis is one of the main ways to obtain sugars from biomass, but most of the polysaccharide-degrading enzymes are poorly efficient on LCB and cellulases with higher performances are required. In this study, we designed a chimeric protein by adding the carbohydrate binding module (CBM) of the cellulosomal enzyme CtLic26A-Cel5E (endoglucanase H or CelH) from Clostridium (Ruminiclostridium) thermocellum to the C-terminus of Dtur CelA, an interesting hyperthermostable endoglucanase from Dictyoglomus turgidum. The activity and binding rate of both native and chimeric enzyme were evaluated on soluble and insoluble polysaccharides. The addition of a CBM resulted in a cellulase with enhanced stability at extreme pHs, higher affinity and activity on insoluble cellulose.
木质纤维素生物质 (LCB) 是一种低成本且丰富的可发酵糖来源。酶解是从生物质中获得糖的主要方法之一，但大多数多糖降解酶对 LCB 的效率较低，需要更高性能的纤维素酶。在这项研究中，我们通过在来自梭菌属（产热梭菌）的纤维二糖结合模块（CBM）的末端添加 Dtur CelA 的 C 末端设计了一种嵌合蛋白，Dtur CelA 是一种有趣的耐热内切葡聚糖酶 Dictyoglomus turgidum。我们评估了天然酶和嵌合酶在可溶性和不溶性多糖上的活性和结合率。添加 CBM 可使纤维素酶在极端 pH 值下具有更高的稳定性、对不溶性纤维素具有更高的亲和力和活性。
