Abstract
Product inhibition by cellobiose decreases the rate of enzymatic cellulose degradation. The optimal reaction conditions for two Emericella (Aspergillus) nidulans-derived cellobiohydrolases I and II produced in Pichia pastoris were identified as CBHI: 52 °C, pH 4.5–6.5, and CBHII: 46 °C, pH 4.8. The optimum in a mixture of the two was 50 °C, pH 4.9. An almost fourfold increase in enzymatic hydrolysis yield was achieved with intermittent product removal of cellobiose with membrane filtration (2 kDa cut-off): The conversion of cotton cellulose after 72 h was ~19 % by weight, whereas the conversion in the parallel batch reaction was only ~5 % by weight. Also, a synergistic effect, achieving ~27 % substrate conversion, was obtained by addition of endo-1,4-β-d-glucanase. The synergistic effect was only obtained with product removal. By using pure, monoactive enzymes, the work illustrates the profound gains achievable by intermittent product removal during cellulose hydrolysis.
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The authors thank Dr. Markus Klinger Dupont-Danisco for the cotton linter and acknowledge Tarbiat Modares University and the Ministry of Science, Research and Technology of Iran for their financial support.
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Gavlighi, H.A., Meyer, A.S. & Mikkelsen, J.D. Enhanced enzymatic cellulose degradation by cellobiohydrolases via product removal. Biotechnol Lett 35, 205–212 (2013). https://doi.org/10.1007/s10529-012-1067-4
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DOI: https://doi.org/10.1007/s10529-012-1067-4