Pristavka A A, Kodituvakku P A, Kozlov Iu P, Zacchi G, Berezin I V, Rabinovich M L
Bach Institute of Biochemistry, Russian Academy of Sciences, Moscow, Russia.
Prikl Biokhim Mikrobiol. 2000 Mar-Apr;36(2):122-30.
A laboratory reactor equipped with a screw press was used for hydrolysis of steam-SO2 exploded willow Salix caprea by a composition of Trichoderma reesei and Aspergillus foetidus enzyme preparations at high substrate concentrations. Optimal conditions providing the maximal volume of hydrolysis syrup with maximal sugar concentrations were determined. Two different hydrolysis procedures were developed in order to exclude initial washing of steam-pretreated plant raw material by large volumes of water, which is necessary to eliminate the inhibitory effect of explosion by-products on enzymatic hydrolysis. The first procedure included a one-hour-long enzymatic prehydrolysis of the substrate, then separation of sugar syrup containing 40-60 g/l of glucose, 20-25 g/l of xylose, and up to 10% of disaccharides, as well as up to 35% of the initial enzymatic activity, then addition of a diluted acetate buffer (pH 4.5), and subsequent hydrolysis of the substrate by the adsorbed enzymes leading to the final accumulation of up to 140 g/l glucose and up to 15 g/l xylose. In the second scenario, the exploded willow was initially adjusted by alkali to pH 4.5 and then hydrolyzed directly by added enzymes for 24 hours. This procedure resulted in a nearly total polysaccharide hydrolysis and accumulation of up to 170 g/l glucose and 20 g/l xylose. The reasons of inhibition of enzymatic hydrolysis are discussed.
使用配备螺旋压榨机的实验室反应器,在高底物浓度下,用里氏木霉和臭曲霉酶制剂的组合物对蒸汽 - SO₂ 爆碎的柳树(Salix caprea)进行水解。确定了能提供最大体积且糖浓度最高的水解糖浆的最佳条件。开发了两种不同的水解程序,以排除大量水洗蒸汽预处理的植物原料,而水洗对于消除爆碎副产物对酶水解的抑制作用是必要的。第一种程序包括对底物进行一小时的酶预水解,然后分离出含有40 - 60 g/l葡萄糖、20 - 25 g/l木糖以及高达10%二糖且酶活性高达初始活性35%的糖浆,接着添加稀释的醋酸盐缓冲液(pH 4.5),随后吸附的酶对底物进行水解,最终积累高达140 g/l葡萄糖和高达15 g/l木糖。在第二种情况下,将爆碎的柳树先用碱调节至pH 4.5,然后直接用添加的酶水解24小时。该程序导致几乎完全的多糖水解,并积累高达170 g/l葡萄糖和20 g/l木糖。文中讨论了酶水解受抑制的原因。