Laboratory of Insect Biochemistry and Physiology, Oswaldo Cruz Institute, Avenida Brasil 4365, Leônidas Deane Building Room 207, Rio de Janeiro, 21040-360, Brazil.
Biotechnol Biofuels. 2011 Nov 14;4:51. doi: 10.1186/1754-6834-4-51.
The description of new hydrolytic enzymes is an important step in the development of techniques which use lignocellulosic materials as a starting point for fuel production. Sugarcane bagasse, which is subjected to pre-treatment, hydrolysis and fermentation for the production of ethanol in several test refineries, is the most promising source of raw material for the production of second generation renewable fuels in Brazil. One problem when screening hydrolytic activities is that the activity against commercial substrates, such as carboxymethylcellulose, does not always correspond to the activity against the natural lignocellulosic material. Besides that, the macroscopic characteristics of the raw material, such as insolubility and heterogeneity, hinder its use for high throughput screenings.
In this paper, we present the preparation of a colloidal suspension of particles obtained from sugarcane bagasse, with minimal chemical change in the lignocellulosic material, and demonstrate its use for high throughput assays of hydrolases using Brazilian termites as the screened organisms.
Important differences between the use of the natural substrate and commercial cellulase substrates, such as carboxymethylcellulose or crystalline cellulose, were observed. This suggests that wood feeding termites, in contrast to litter feeding termites, might not be the best source for enzymes that degrade sugarcane biomass.
新水解酶的描述是利用木质纤维素材料作为燃料生产起点的技术发展的重要步骤。甘蔗渣经过预处理、水解和发酵，在几家试验性精炼厂中生产乙醇，是巴西生产第二代可再生燃料最有前途的原料来源。在筛选水解活性时存在一个问题，即对商业底物（如羧甲基纤维素）的活性并不总是与天然木质纤维素材料的活性相对应。此外，原材料的宏观特性，如不溶性和异质性，阻碍了其用于高通量筛选。
本文介绍了一种从甘蔗渣中获得的颗粒胶体悬浮液的制备方法，该方法对木质纤维素材料的化学变化最小，并展示了其在使用巴西白蚁作为筛选生物的高通量水解酶测定中的应用。
观察到天然底物和商业纤维素酶底物（如羧甲基纤维素或结晶纤维素）的使用之间存在重要差异。这表明，与枯枝落叶取食白蚁相比，木材取食白蚁可能不是降解甘蔗生物质的酶的最佳来源。
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