Enhanced Enzymatic Hydrolysis of High-Solids Content Corncobs by a Lytic Polysaccharide Monooxygenase from Podospora anserina S Mat+ for Valuable Monosaccharides-猫眼课题宝

DOI：

10.1021/acssuschemeng.3c02691

发表时间：

2023-06

期刊：

ACS Sustainable Chemistry & Engineering

影响因子：

通讯作者：

中科院分区：

其他

文献类型：

作者：

Xue Cai;Jing Hua;Zhengyou Lin;Chen-Yang Sun;Chang-Hui Hu;Xiao‐Jian Zhang;Ji-dong Shen;Hai-yan Zhou;Hong-Yan Wang;Kai-Qian Chen;De-Shui Chen;Xin-Ping Cheng;Mian Li;Zhi-qiang Liu;Yuguo Zheng

多糖单加氧酶（LPMO）可以提高纤维素酶的活性，促进玉米芯纤维素的高效利用。为此，通过硅基筛选，从anserinaS mat+ (podospora anserinaS mat+)中分离出一种新的LPMO，在大肠杆菌bl21 （DE3）中异种表达，并对其生化特性进行了研究。为了降低生产成本，使用伴侣和定点诱变来提高蛋白质的溶解度，产量提高了8%。通过单因素实验，确定高固含量25%玉米芯、60℃、pH 5.5、palpmo与纤维素酶比1:3（总蛋白浓度为12 mg/g）、添加0.5 mmol/L Mn2+的最佳水解条件为葡萄糖产率129.8 g/L。与单独使用纤维素酶相比，这一结果提高了24%，在相同水解程度下，商用纤维素酶的使用量减少了33%。将高固含量玉米芯降解后的水解产物进一步通过葡萄糖异构酶和-allulose 3- epimase以2:1的比例催化生产有价值的allulose，果糖的产率为56.22 g/L， allulose的产率为20.6 g/L。PaLPMO和纤维素酶复合物显著提高了玉米芯渣的降解效率，减少了纤维素酶的使用，降低了生产成本。这些研究结果有助于开发玉米芯综合利用，为可持续、高效地利用能源和资源铺平道路。

Lytic polysaccharide monooxygenase (LPMO) has been found to enhance cellulase activity and promote efficient utilization of corncob cellulose. To this end, a novel LPMO fromPodospora anserinaS mat+ (PaLPMO) throughin silicoscreening was heterologously expressed inEscherichia coliBL21(DE3) and its biochemical properties were investigated. To reduce production costs, chaperones and site-directed mutagenesis were employed to enhance the protein’s solubility, yielding an 8% increase. Through single-factor experiments, the optimal hydrolysis condition for the highest glucose yield of 129.8 g/L was identified as 25% high-solids content of corncob at 60 °C and pH 5.5 using a PaLPMO-to-cellulase ratio of 1:3 (total protein concentration of 12 mg/g) and with the addition of 0.5 mmol/L Mn2+. This result represented a 24% improvement compared to cellulase alone and a 33% reduction in commercial cellulase usage with the same degree of hydrolysis. The hydrolysis products of high-solids content corncob degradation were further used to produce valuabled-allulose through glucose isomerase andd-allulose 3-epimerase catalysis in a ratio of 2:1, with a yield of 56.22 g/L fructose and 20.6 g/Ld-allulose. The PaLPMO and cellulase complex significantly improved the degradation efficiency of corncob residue and reduced cellulase usage in lowering the production cost. These findings contribute to developing comprehensive corncob utilization and pave the way for sustainable and efficient energy and resource utilization.

Enhanced Enzymatic Hydrolysis of High-Solids Content Corncobs by a Lytic Polysaccharide Monooxygenase from Podospora anserina S Mat+ for Valuable Monosaccharides

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