Department of Biotechnology, University of São Paulo, School of Engineering of Lorena, Estrada Municipal do Campinho- Caixa,, Postal 116 12,602,810, Lorena/SP, Brazil.
Biotechnol Biofuels. 2013 Jan 16;6(1):4. doi: 10.1186/1754-6834-6-4.
Diminishing supplies of fossil fuels and oil spills are rousing to explore the alternative sources of energy that can be produced from non-food/feed-based substrates. Due to its abundance, sugarcane bagasse (SB) could be a model substrate for the second-generation biofuel cellulosic ethanol. However, the efficient bioconversion of SB remains a challenge for the commercial production of cellulosic ethanol. We hypothesized that oxalic-acid-mediated thermochemical pretreatment (OAFEX) would overcome the native recalcitrance of SB by enhancing the cellulase amenability toward the embedded cellulosic microfibrils.
OAFEX treatment revealed the solubilization of hemicellulose releasing sugars (12.56 g/l xylose and 1.85 g/l glucose), leaving cellulignin in an accessible form for enzymatic hydrolysis. The highest hydrolytic efficiency (66.51%) of cellulignin was achieved by enzymatic hydrolysis (Celluclast 1.5 L and Novozym 188). The ultrastructure characterization of SB using scanning electron microscopy (SEM), atomic force microscopy (AFM), Raman spectroscopy, Fourier transform-near infrared spectroscopy (FT-NIR), Fourier transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD) revealed structural differences before and after OAFEX treatment with enzymatic hydrolysis. Furthermore, fermentation mediated by C. shehatae UFMG HM52.2 and S. cerevisiae 174 showed fuel ethanol production from detoxified acid (3.2 g/l, yield 0.353 g/g; 0.52 g/l, yield, 0.246 g/g) and enzymatic hydrolysates (4.83 g/l, yield, 0.28 g/g; 6.6 g/l, yield 0.46 g/g).
OAFEX treatment revealed marked hemicellulose degradation, improving the cellulases' ability to access the cellulignin and release fermentable sugars from the pretreated substrate. The ultrastructure of SB after OAFEX and enzymatic hydrolysis of cellulignin established thorough insights at the molecular level.
化石燃料供应减少和石油泄漏促使人们探索可从非粮/饲料基底物生产的替代能源。由于其丰富的资源,甘蔗渣(SB)可能成为第二代生物燃料纤维素乙醇的模型底物。然而,SB 的高效生物转化仍然是纤维素乙醇商业化生产的一个挑战。我们假设草酸介导的热化学预处理(OAFEX)可以通过增强纤维素酶对嵌入的纤维素微纤维的可及性来克服 SB 的天然顽固性。
OAFEX 处理揭示了半纤维素的溶解,释放出糖(12.56 g/L 木糖和 1.85 g/L 葡萄糖),使纤维素木质素以可接近的形式用于酶解。通过酶解(Celluclast 1.5 L 和 Novozym 188)实现了纤维素木质素的最高水解效率(66.51%)。利用扫描电子显微镜(SEM)、原子力显微镜(AFM)、拉曼光谱、傅里叶变换近红外光谱(FT-NIR)、傅里叶变换红外光谱(FTIR)和 X 射线衍射(XRD)对 SB 的超微结构进行了表征,发现了 OAFEX 处理前后的结构差异,以及酶解处理后的结构差异。此外,C. shehatae UFMG HM52.2 和 S. cerevisiae 174 通过发酵从解毒酸(3.2 g/L,产率 0.353 g/g;0.52 g/L,产率 0.246 g/g)和酶解产物(4.83 g/L,产率 0.28 g/g;6.6 g/L,产率 0.46 g/g)中生产燃料乙醇。
OAFEX 处理显著降解了半纤维素,提高了纤维素酶对预处理底物中纤维素木质素的可及性,并释放出可发酵糖。SB 经 OAFEX 处理和纤维素木质素酶解后的超微结构在分子水平上建立了全面的见解。