Sharma Gaurav, Singh Varinder, Raheja Yashika, Chadha Bhupinder Singh
Department of Microbiology, Guru Nanak Dev University, Amritsar, 143005 Punjab India.
3 Biotech. 2024 Jun;14(6):168. doi: 10.1007/s13205-024-04013-7. Epub 2024 May 30.
The lignocellulolytic accessory enzyme, Feruloyl esterase C (FE_5DR), encoded in the genome of thermotolerant was successfully cloned and heterologously expressed in . The expressed FE_5DR was purified using UNOsphere™ Q anion exchange chromatography column, exhibiting a homogeneous band of ~ 39 kDa. Its optimum temperature was determined to be 60 °C, with an optimal pH of 6.0. Additionally, the enzyme activity of FE_5DR was significantly enhanced by preincubation in a buffer containing Mg, Cu and Ca metal ions. Enzyme kinetic parameters, computed from double reciprocal Lineweaver-Burk plots, yielded observed V and K values of 0.758 U/mg and 0.439 mM, respectively. Furthermore, the potential of custom-made cocktails comprising FE_5DR and benchmark cellulase derived from the developed mutant strain of MAN 40, as well as the biorefinery-relevant lignocellulolytic enzyme Cellic CTec 3, resulted in improved saccharification of unwashed acid pretreated (UWAP) rice straw slurry and mild alkali deacetylated (MAD) rice straw when compared to benchmark MAN 40 and Cellic CTec 3.
The online version contains supplementary material available at 10.1007/s13205-024-04013-7.
在耐热菌基因组中编码的木质纤维素分解辅助酶阿魏酸酯酶C（FE_5DR）已成功克隆并在[具体表达宿主]中进行了异源表达。使用UNOsphere™ Q阴离子交换色谱柱对表达的FE_5DR进行纯化，显示出一条约39 kDa的均一蛋白条带。其最适温度确定为60°C，最适pH为6.0。此外，在含有镁、铜和钙金属离子的缓冲液中预孵育可显著提高FE_5DR的酶活性。根据双倒数Lineweaver - Burk图计算的酶动力学参数，观察到的V和K值分别为0.758 U/mg和0.439 mM。此外，与基准菌株MAN 40和纤维素酶Cellic CTec 3相比，由[具体菌株]的突变菌株开发的包含FE_5DR和基准纤维素酶的定制酶混合物以及与生物精炼相关的木质纤维素分解酶Cellic CTec 3，可提高未洗涤酸预处理（UWAP）稻草浆和轻度碱脱乙酰化（MAD）稻草的糖化效果。
在线版本包含可在10.1007/s13205 - 024 - 04013 - 7获取的补充材料。
