Biological Resources and Post-harvest Division, Japan International Research Center for Agricultural Sciences (JIRCAS), 1-1 Ohwashi, Tsukuba, Ibaraki 303-8686, Japan.
Bioresour Technol. 2013 Feb;130:424-30. doi: 10.1016/j.biortech.2012.12.059. Epub 2012 Dec 20.
This report describes a novel recycling method utilizing a combination of Clostridium thermocellum cellulosomes and Thermoanaerobacter brockii β-glucosidase (CglT). To recover cellulosomes and CglT through re-binding to additional cellulose, a chimeric CBM3-CglT was created by fusing carbohydrate binding module (CBM3) from the scaffolding protein CipA into the N-terminal region of CglT. When a recycling test using cellulosomes and CBM3-CglT was performed on microcrystalline cellulose, the process was capable of 4 rounds of recycling (1%w/vcellulose/round). Although irreversible absorption of cellulosomes and CBM3-CglT into the residues was observed when ammonia-pretreated rice straw and delignified rice straw was used as substrates, a maximum of 2 and 4 recycling rounds (1%w/vglucan/round) were achieved, respectively, consistent with a 70% saccharification rate. This novel recycling method using cellulosomes and CBM3-CglT has great potential as an effective lignocellulose degradation system.
本报告描述了一种新颖的回收方法，利用了嗜热纤维梭菌细胞表面展示纤维素酶和热厌氧菌β-葡萄糖苷酶（CglT）的组合。为了通过重新结合到额外的纤维素上来回收细胞表面展示纤维素酶和 CglT，通过将碳水化合物结合模块（CBM3）从支架蛋白 CipA 融合到 CglT 的 N 端区域，创建了一种嵌合 CBM3-CglT。当在微晶纤维素上进行使用细胞表面展示纤维素酶和 CBM3-CglT 的回收测试时，该过程能够进行 4 轮回收（每轮 1%w/v 纤维素）。尽管在用氨预处理的稻草和脱木质素的稻草作为底物时观察到细胞表面展示纤维素酶和 CBM3-CglT 不可逆地吸收到残留物中，但仍分别实现了最大 2 和 4 轮回收（每轮 1%w/v 葡聚糖），与 70%的糖化率一致。这种使用细胞表面展示纤维素酶和 CBM3-CglT 的新型回收方法具有作为有效木质纤维素降解系统的巨大潜力。
