Isolation of Paraclostridium CR4 from sugarcane bagasse and its evaluation in the bioconversion of lignocellulosic feedstock into hydrogen by monitoring cellulase gene expression.

Bioconversion of sugarcane bagasse (SCB) into hydrogen (H) and organic acids was evaluated using a biomolecular approach to monitor the quantity and expression of the cellulase (Cel) gene. Batch reactors at 37 °C were operated with Paraclostridium sp. (10% v/v) and different substrates (5 g/L): glucose, cellulose and SCB in natura and pre-heat treated and hydrothermally. H production from glucose was 162.4 mL via acetic acid (2.9 g/L) and 78.4 mL from cellulose via butyric acid (2.9 g/L). H production was higher in hydrothermally pretreated SCB reactors (92.0 mL), heat treated (62.5 mL), when compared to in natura SCB (51.4 mL). Butyric acid (5.8, 4.9 and 4.0 g/L) was the main acid observed in hydrothermally, thermally pretreated, and in natura SCB, respectively. In the reactors with cellulose and reactors with hydrothermally pretreated SCB, the Cel gene copy number 3 and 2 log were higher, respectively, during the stage of maximum H production rate, when compared to the initial stage. Differences in Cel gene expression were observed according to the concentration of soluble sugars in the reaction medium. That is, there was no gene expression at the initial phase of the experiment using SCB with 2.6 g/L of sugars and increase of 2.2 log in gene expression during the phases with soluble sugars of <1.4 g/L.