how recombinant swollenin from kluyveromyces lactis affects cellulosic substrates and accelerates their hydrolysis如何重组swollenin克鲁维酵母菌属lactis影响纤维素基质和加速水解.pdfVIP

Jäger et al. Biotechnology for Biofuels 2011, 4:33 /content/4/1/33 RESEARCH Open Access How recombinant swollenin from Kluyveromyces lactis affects cellulosic substrates and accelerates their hydrolysis 1† 2† 1 3 3 2 Gernot Jäger , Michele Girfoglio , Florian Dollo , Roberto Rinaldi , Hans Bongard , Ulrich Commandeur , Rainer Fischer2,4, Antje C Spiess5 and Jochen Büchs1* Abstract Background: In order to generate biofuels, insoluble cellulosic substrates are pretreated and subsequently hydrolyzed with cellulases. One way to pretreat cellulose in a safe and environmentally friendly manner is to apply, under mild conditions, non-hydrolyzing proteins such as swollenin - naturally produced in low yields by the fungus Trichoderma reesei. To yield sufficient swollenin for industrial applications, the first aim of this study is to present a new way of producing recombinant swollenin. The main objective is to show how swollenin quantitatively affects relevant physical properties of cellulosic substrates and how it affects subsequent hydrolysis. Results: After expression in the yeast Kluyveromyces lactis, the resulting swollenin was purified. The adsorption parameters of the recombinant swollenin onto cellulose were quantified for the first time and were comparable to those of individual cellulases from T. reesei. Four different insoluble cellulosic substrates were then pretreated with swollenin. At first, it could be qualitatively shown by macroscopic evaluation and microscopy that swollenin caused deagglomeration of bigger cellulose agglomerates as well as dispersion of cellulose microfibrils (amorphogenesis). Afterwards, the effects of swollenin on cellulose particle size, maximum ce