Department of Chemistry, National Changhua University of Education, Changhua, Taiwan.
Biotechnol Lett. 2011 Apr;33(4):777-82. doi: 10.1007/s10529-010-0500-9. Epub 2010 Dec 25.
The synergism between the endo/exocellulase, Cel9A, and β-glucosidase (βgl) of Thermobifida fusca was investigated. Wild type βgl or S319C, a βgl mutant with significantly improved cellobiase activity, were added to Cel9A. Both wild type and mutant βgl enhanced the Cel9A hydrolysis of carboxymethyl cellulose (CMC) and filter paper by 50-100% compared to Cel9A alone. No enhancement occurred with addition of E388A, an inactive form of βgl. HPLC analysis showed that, with Cel9A alone, the resulting hydrolysate of glucose and cellobiose contained about half glucose; after addition of equimolar amounts of either wild type βgl or mutant S319C to Cel9A, the hydrolysate contained more than 85% glucose. βgl thus acted synergistically with Cel9A by converting cello-oligomers to glucose; this reduced the soluble sugar accumulation during hydrolysis of cellulose.
研究了嗜热纤维梭菌的内切/外切纤维素酶、Cel9A 和 β-葡萄糖苷酶(βgl)之间的协同作用。向 Cel9A 中添加野生型βgl 或 S319C,这是一种细胞二糖酶活性显著提高的βgl 突变体。与单独使用 Cel9A 相比,野生型和突变型 βgl 均将 Cel9A 对羧甲基纤维素(CMC)和滤纸的水解提高了 50-100%。添加 E388A(βgl 的无活性形式)不会发生增强作用。HPLC 分析表明,单独使用 Cel9A 时,葡萄糖和纤维二糖的水解产物中约含有一半的葡萄糖;向 Cel9A 中添加等摩尔量的野生型βgl 或突变型 S319C 后,水解产物中含有超过 85%的葡萄糖。因此,βgl 通过将纤维寡糖转化为葡萄糖与 Cel9A 协同作用;这减少了纤维素水解过程中可溶性糖的积累。