High throughput screening of hydrolytic enzymes from termites using a natural substrate derived from sugarcane bagasse.

利用源自甘蔗渣的天然底物对白蚁水解酶进行高通量筛选

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作者:Lucena Severino A, Lima Leile S, Cordeiro Luís Sa Jr, Sant'anna Celso, Constantino Reginaldo, Azambuja Patricia, de Souza Wanderley, Garcia Eloi S, Genta Fernando A
BACKGROUND: 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. RESULTS: 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. CONCLUSIONS: 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.

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