Abstract: Disclosed is a process for simply preparing cell extracts for use as a catalyst of cell-free protein synthesis by centrifugation, which improves cost effectiveness and productivity of cell-free protein synthesis. Specifically, a conventional process for preparing cell extracts comprises the complicated steps, i.e. cell culture, cell lysis, high-speed centrifugation, pre-incubation, dialysis and the like. In comparison, the cell lysate just obtained by centrifugation is directly applied to protein synthesis, thereby providing higher producibility and more consistent productivity of protein than the conventional process. Further, the cell extracts are prepared by the simple process to reduce the protein production cost and time by about 60% and about 80%, respectively.

Abstract: The present invention relates to a method for synthesizing a protein in a cell-free synthesis system, which comprises the steps of: (a) amplifying a DNA molecule encoding the protein to prepare linear DNA molecules with a stem and loop sequence at their terminus, wherein the stem and loop sequence comprises a nucleotide sequence inducing the formation of a stem and loop structure at the 3?-end of transcripts of the amplified DNA molecules to prevent degradation of the transcripts; and (b) contacting the amplified DNA molecules to a lysate of cells with deficient endoribonuclease E activity, thereby producing the protein in the cell-free synthesis system.

Abstract: The present invention relates to methods of production of the completely post-translationally modified protein by combination of cell-free protein synthesis and cell-free co- and post-translational modification. Previous cell-free protein synthesis system has only been capable of producing partially post-translationally modified protein but the present invention employs a co- and post-translational modification machinery that produces completely post-translationally modified protein.

Abstract: The present invention relates to methods of production of the completely post-translationally modified protein by combination of cell-free protein synthesis and cell-free co- and post-translational modification. Previous cell-free protein synthesis system has only been capable of producing partially post-translationally modified protein but the present invention employs a co- and post-translational modification machinery that produces completely post-translationally modified protein.

Abstract: The present invention discloses modified muteins of erythropoietin (EPO) with increased biological activity derived from in vitro or in vivo expression system of microorganism exhibited a prolonged circulating half-life compared to unmodified muteins of EPO produced from microorganisms, and novel methods of preparing the selectively modified muteins of EPO.

Abstract: A metabolite, e.g., ethanol, is continuously produced from low cost carbohydrate substrates by a process which comprises pulverizing the carbohydrate substrate; liquefying and saccharifying the pulverized substrate; continuously fermenting the lique-saccharified substrate in a fermentor equipped with a moving filter, in the presence of flocculent biological cells maintained at a concentration ranging from 90 to 160 g/l by using the moving filter and a culture medium to produce a fermentation product mixture; and recovering the desired metabolite from the fermentation product mixture.

Abstract: A method for producing protein in a cell-free system uses cell-free extract required for protein production, extracted from a biological cell. According to the present invention, a wide range of proteins can be produced economically and efficiently even in a semi-continuous operation mode, by regenerating energy source, increasing operation time of the reactor with porous solid material and using selective protein isolating means having high specific affinity to a desired protein. Thus, problems appearing in the living cell (in vivo) system due to the absence of post-translational processing can be avoided and the total production amount can be greatly increased due to prolonged operation time of the reactor and increased protein productivity.