期刊
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
卷 22, 期 1, 页码 -出版社
MDPI
DOI: 10.3390/ijms22010276
关键词
GH6 cellobiohydrolase; AA9 LPMO; lignocellulose hydrolysis; bioethanol
资金
- Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) [2016/190950-0, 2018/11231-7]
- Conselho Nacional de Desenvolvimento Cientifico e Tecnologico [CNPq: 425465/2016-0]
- Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior-Brasil (CAPES) [001]
The cellulose-active enzymes secreted by Aspergillus fumigatus play a crucial role in degrading lignocellulosic biomass, although some combinations of enzymes may inhibit each other. However, adding these enzymes to a cellulase cocktail can enhance the saccharification of complex substrates and increase the yield of fermentable sugars.
Cellulose is the most abundant polysaccharide in lignocellulosic biomass, where it is interlinked with lignin and hemicellulose. Bioethanol can be produced from biomass. Since breaking down biomass is difficult, cellulose-active enzymes secreted by filamentous fungi play an important role in degrading recalcitrant lignocellulosic biomass. We characterized a cellobiohydrolase (AfCel6A) and lytic polysaccharide monooxygenase LPMO (AfAA9_B) from Aspergillus fumigatus after they were expressed in Pichia pastoris and purified. The biochemical parameters suggested that the enzymes were stable; the optimal temperature was similar to 60 degrees C. Further characterization revealed high turnover numbers (k(cat) of 147.9 s(-1) and 0.64 s(-1), respectively). Surprisingly, when combined, AfCel6A and AfAA9_B did not act synergistically. AfCel6A and AfAA9_B association inhibited AfCel6A activity, an outcome that needs to be further investigated. However, AfCel6A or AfAA9_B addition boosted the enzymatic saccharification activity of a cellulase cocktail and the activity of cellulase Af-EGL7. Enzymatic cocktail supplementation with AfCel6A or AfAA9_B boosted the yield of fermentable sugars from complex substrates, especially sugarcane exploded bagasse, by up to 95%. The synergism between the cellulase cocktail and AfAA9_B was enzyme- and substrate-specific, which suggests a specific enzymatic cocktail for each biomass by up to 95%. The synergism between the cellulase cocktail and AfAA9_B was enzyme- and substrate-specific, which suggests a specific enzymatic cocktail for each biomass.
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