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通过工程化草酸青霉改造纤维素酶系统生产纤维素纳米晶体。

Modification of a cellulase system by engineering Penicillium oxalicum to produce cellulose nanocrystal.

机构信息

State Key Laboratory of Microbial Technology, Shandong University, No.72, Binhai Road, Qingdao City, 266237, Shandong, China.

State Key Laboratory of Microbial Technology, Shandong University, No.72, Binhai Road, Qingdao City, 266237, Shandong, China.

出版信息

Carbohydr Polym. 2020 Apr 15;234:115862. doi: 10.1016/j.carbpol.2020.115862. Epub 2020 Jan 16.

Abstract

In this study, three endoglucanases (EGs; Cel7B, Cel5B, and Cel12A), one cellobiohydrolase (CBH), and two auxiliary proteins (swollenin: SWO1 and SWO4) were used to hydrolyze microcrystalline cellulose (MCC) for cellulose nanocrystal (CNC) preparation. The mixture experiment of the three EGs showed that high CNC yield was obtained when the ratio of Cel7B and Cel5B is 1:1.11 (protein weight). Moreover, the addition of CBH (1 mg/g) and SWO1 from Trichoderma reesei effectively increased the yield of CNC. On the basis of the results, the cellulase-producing strain of Penicillium oxalicum M12 was engineered to improve its cellulase system. An engineered strain of cEES performed well in CNC preparation. CNC with a yield of 11.79 % and a crystallinity of 83.85 % were produced using the crude enzyme from cEES as a means to hydrolyze MCC, and the size and shape of CNC were uniform and fusiform.

摘要

在这项研究中,使用了三种内切葡聚糖酶(EGs;Cel7B、Cel5B 和 Cel12A)、一种纤维二糖水解酶(CBH)和两种辅助蛋白(膨胀素:SWO1 和 SWO4)来水解微晶纤维素(MCC)以制备纤维素纳米晶体(CNC)。三种 EGs 的混合物实验表明,当 Cel7B 和 Cel5B 的比例为 1:1.11(蛋白重量)时,可获得高 CNC 产率。此外,添加来自里氏木霉的 CBH(1mg/g)和 SWO1 可有效提高 CNC 的产率。在此基础上,对产纤维素酶的草酸青霉 M12 进行了工程改造,以改善其纤维素酶系统。经过工程改造的 cEES 菌株在 CNC 制备方面表现良好。使用 cEES 的粗酶水解 MCC 可生产出产率为 11.79%、结晶度为 83.85%的 CNC,其尺寸和形状均匀且呈梭形。

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