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基于碱性预处理和响应面法的甘蔗梢重组酶糖化与发酵

Alkaline pretreatment and response surface methodology based recombinant enzymatic saccharification and fermentation of sugarcane tops.

作者信息

Chandrakant Khaire Kaustubh, Suryakant Moholkar Vijayanand, Goyal Arun

机构信息

School of Energy Science and Engineering, Indian Institute of Technology Guwahati, Guwahati 781039, Assam, India.

School of Energy Science and Engineering, Indian Institute of Technology Guwahati, Guwahati 781039, Assam, India; Department of Chemical Engineering, Indian Institute of Technology Guwahati, Guwahati 781039, Assam, India.

出版信息

Bioresour Technol. 2021 Dec;341:125837. doi: 10.1016/j.biortech.2021.125837. Epub 2021 Aug 25.

Abstract

In present study, the water-soluble extractives removal prior to alkali pretreatment of sugarcane tops (SCT) was carried out. The solid alkali pretreated SCT (apSCT) recovered on Field-emission scanning electron microscopy (FE-SEM) analysis showed exposure of cellulosic fibres as compared with raw SCT. The analyses of apSCT by Fourier Transform Infrared (FT-IR) Spectroscopy, X-ray diffraction (XRD) and High performance liquid chromatography (HPLC) analysis also confirmed the enhanced cellulose content in apSCT. Optimum conditions for response surface methodology based saccharification of apSCT at 40 °C, 150 rpm were 2.14% (w/v) apSCT loading in citrate-phosphate buffer (50 mM, pH 6.0), recombinant hydrolytic enzymes (from Clostridium/Hungateiclostridium thermocellum) loading for endo-1,4-β-glucanase (CtCel8A) = 213.2 U/g, cellobiohydrolase (CtCBH5A) = 272.5 U/g and β-glucosidase (HtBg1) = 299.8 U/g for 49.2 h. Under optimized saccharification conditions, the total reducing sugar yield was 265 mg/g (glucose 214 mg/g) of apSCT. Fermentation of produced glucose by S. cerevisiae gave 0.19 g/g glucose of bioethanol.

摘要

在本研究中,对甘蔗梢(SCT)进行了碱预处理前的水溶性提取物去除操作。场发射扫描电子显微镜(FE-SEM)分析显示,回收的固体碱预处理甘蔗梢(apSCT)与未处理的甘蔗梢相比,纤维素纤维暴露出来。傅里叶变换红外光谱(FT-IR)、X射线衍射(XRD)和高效液相色谱(HPLC)对apSCT的分析也证实了apSCT中纤维素含量增加。基于响应面法在40°C、150 rpm条件下对apSCT进行糖化的最佳条件为:在柠檬酸盐-磷酸盐缓冲液(50 mM,pH 6.0)中,apSCT负载量为2.14%(w/v),重组水解酶(来自嗜热栖热梭菌/亨盖特栖热梭菌)负载量为:内切-1,4-β-葡聚糖酶(CtCel8A)=213.2 U/g,纤维二糖水解酶(CtCBH5A)=272.5 U/g,β-葡萄糖苷酶(HtBg1)=299.8 U/g,反应49.2小时。在优化的糖化条件下,apSCT的总还原糖产量为265 mg/g(葡萄糖214 mg/g)。酿酒酵母对产生的葡萄糖进行发酵,得到0.19 g/g葡萄糖的生物乙醇。

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