Conversion of biomass to ethanol |
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Authors: | Lee Ching-Yi Wen Jingquan Thomas Seqwana Delgass W. Nicholas Grutzner John B. Tsao George T. |
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Affiliation: | (1) Biotechnology Research Laboratory, Kobe Steel Ltd., 1-25-14 Kannondai, 305 Tsukuba, Japan |
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Abstract: | In theEscherichia coli cell-free system, the modification of cell extract can be achieved by preparation of the strains carrying additional property or those being induced with a certain gene expression prior to harvesting. In this study, we analyzed the cell-free system with S30 extract containing T7 RNA polymerases (S30 extract-T7pol) prepared from E.coli BL21(DE3) strain, which includes T7 RNA polymerase from extrinsic genes by IPTG induction, as a model for the improvement of the cell-free system. The fact that a significant degree of mRNA degradation was observed in the cell-free system with S30 extract-T7pol indicates the increase of ribonuclease activity was an unfavorable influence derived from the cell-extract modification process. We also showed that this influence was settled by the addition of an effective ribonuclease inhibitor, such as copper (II) ion, to the reaction mixture. |
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Keywords: | Cell-free system copper ion ribonuclease Escherichia coli protein synthesis |
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