Provided is a method for biosynthesis of (R)-?-hydroxytetradecanoate compounds. The method comprises the following steps of: reacting ?-carbonyl tetradecanoate in a ketoreductase and a glucose dehydrogenase or a fusion enzyme thereof, glucose, NADP.sup.+ and a buffer solution to obtain (R)-?-hydroxytetradecanoate compounds. The method is an enzyme-catalyzed biosynthesis method having simple operations, conventional equipment and an environmentally friendly process, and the prepared product has high purity, high yield and a high ee value.

Geranyl pyrophosphate (GPP) is a key intermediate molecule in the bioproduction of thousands of natural products. Currently, natural products are either cultivated from plants, synthesized via complex chemical synthesis strategies, or through cell-based factories also known as biofoundries. However, in order to replicate the process in a cell free environment, numerous enzymes and cofactors must be utilized making this approach costly and unviable. In order to make this process viable, a new approach was needed that uses fewer enzymes and co-factors. As described herein, the present invention demonstrates that it is possible to create GPP from glycerol through a short and concise biosynthetic pathway outside of the cell.

In one non-limiting aspect, sterilizable reagent materials for diagnostic elements are provided. In other aspects, sterilized elements and techniques for producing the same are disclosed. In one embodiment, a sterilized element includes a chemical detection reagent including at least one component that is sensitive to ionizing radiation. The sterilized element is also mediator-free and the at least one component sensitive to ionizing radiation is present in a functional form in a proportion of 80% based on the total amount of the respective component in the diagnostic element before sterilization. In certain aspects, the at least one component sensitive to ionizing radiation includes one or both of an enzyme and a coenzyme. Other aspects include, but are not limited to, unique methods, techniques, products, systems and devices involving sterilizable reagent materials or sterilized elements.

Provided is a process for the preparation of L-methionine in an enzymatic reaction utilizing dimethyl disulfide (DMDS) a precursor of L-methionine, and an organic reducing compound. In the process, methyl mercaptan can be formed by the enzymatic hydrogenolysis of the DMDS.

In some embodiments, the present invention provides a protein comprising amino acids in the following sequence L(X).sub.n=14X.sup.3(X).sub.n=1X.sup.1(X).sub.n=1E(X).sub.n=4P(X).sub.n=1NR(X).sub.n=3S(X).sub.n=4D(X).sub.n=2G(X).sub.n=7Y(X).sub.n=4Y (X).sub.n=32-34X.sup.2, wherein each X independently represents any naturally occurring amino acid residue and n indicates the number of amino acid residues represented by the respective parenthetical at that position, wherein: a) X1 is selected from the group consisting of S, C, T, M, V, Y, N, P, L, G, Q, A, I, D, W, H, or E, wherein if X.sup.1 is L, H or V, then X.sup.3 is D; and/or b) X.sup.2 is selected from the group consisting of H, L, S or V. In some embodiments, the present invention also provides a protein comprising amino acids in the sequence set forth by SEQ ID NO: 38 or SEQ ID NO: 39, except that: the amino acid at position 406 is an amino acid other than F; and/or the amino acid at position 474 is an amino acid other than N.

The present invention provides a glucose dehydrogenase having an improved specific activity. A polypeptide comprising an amino acid sequence having such a structure that an amino acid residue at position-578 is substituted by a valine residue or a phenylalanine residue in the amino acid sequence represented by SEQ ID NO: 1, and a variant of the polypeptide have an improved glucose dehydrogenase activity.

The present invention provides a process for obtaining n+a oxidation and reduction products from a mixture of n sugars selected from the group consisting of C5 and C6 sugars, wherein n is at least 2 and a is at least 1, wherein at least two of the sugars in the mixture are present at a non-equimolar ratio to each other, wherein, in a first stage, at least one of the sugars which are present at a non-equimolar ratio to each other is oxidized enzymatically and, at the same time, at least one of the other sugars present at a non-equimolar ratio to each other is reduced enzymatically, and wherein, in the first stage, a portion of at least one of the sugars present at a non-equimolar ratio to each other is not converted, and which is characterized in that, in at least a second stage, at least a portion of the sugar not converted in the first stage is oxidized enzymatically by half and, respectively, is reduced enzymatically by the remaining half.

Provided are a recombinant microorganism including a genetic modification that increases a pyruvate, phosphate dikinase activity, a method of producing cellulose using the same, and a method of producing a microorganism having enhanced cellulose productivity.

Disclosed are a phosphinothricin dehydrogenase mutant, a recombinant bacterium and a one-pot multi-enzyme synchronous directed evolution method. The phosphinothricin dehydrogenase mutant, with an amino acid sequence as shown in SEQ ID No.1, is obtained by mutating alanine at position 164 to glycine, arginine at position 205 to lysine, and threonine at position 332 to alanine in a phosphinothricin dehydrogenase derived from Pseudomonas fluorescens. The recombinant bacterium is obtained by introducing a gene encoding the phosphinothricin dehydrogenase mutant into a host cell. The host cell can also incorporate a gene encoding a glucose dehydrogenase or a gene encoding a formate dehydrogenase to undergo synchronous directed evolution to achieve double gene overexpression. The one-pot multi-enzyme synchronous directed evolution method of the present invention can screen recombinant bacteria with greatly improved activity. Compared with other catalysis processes such as the transaminase method, the method for preparing L-PPT of the present invention features relatively simple process, high conversion of raw materials of up to 100%, and high stereo selectivity.

Described are methods for generating a reporter molecule in response to a target analyte in a cell-free system. A synthetic biological circuit is used to modify the level of the reporter molecule in response to the presence of the target analyte. The reporter molecule may be glucose or another molecule readily detected using a device such as glucose monitor or other portable sensor. Also provided are kits comprising a cell-free system with a synthetic biological circuit that generates or consumes a reporter molecule in response to a target analyte.