The present disclosure provides engineered ketoreductase enzymes having improved properties as compared to a naturally occurring wild-type ketoreductase enzyme. Also provided are polynucleotides encoding the engineered ketoreductase enzymes, host cells capable of expressing the engineered ketoreductase enzymes, and methods of using the engineered ketoreductase enzymes to synthesize a variety of chiral compounds.

Provided are amino acid sequences of ketoreductase polypeptides that are useful for asymmetrically synthesizing chiral alcohol compounds, its preparation process as well as reaction process under industrial-relevant conditions. Also provided are polynucleotide sequences encoding engineered ketoreductase polypeptides, engineered host cells capable of expressing engineered ketoreductase polypeptides, and methods of producing chiral alcohol compounds using engineered ketoreductase polypeptides. Compared to other enzymes, the engineered ketoreductase polypeptides provided by the invention have better catalytic activity and thermal stability, allowing purification of the enzyme solution by heat treatment, which is advantageous for the production of enzymes and the industrial application of enzymatic reactions. The use of the engineered polypeptides of the present invention greatly simplifies the production process of chiral alcohol compounds, reduces the cost of production and the impact on the environment, and has good industrial application prospects.

A monooxygenase having an amino acid sequence obtained by mutation of the amino acid sequence shown in SEQ ID NO:2 is disclosed. The use of the monooxygenase of the present invention in production of chiral sulfoxide-based drugs has advantages including mild reaction conditions, environmental friendliness, high yield, high optical purity of products, less peroxide products, and the like, and therefore the monooxygenase in the present invention has a good industrial application prospect in the production of proton pump inhibitors for the treatment of gastric ulcers.

A process for the enzymatic regeneration of the redox cofactors NAD.sup.+/NADH and NADP.sup.+/NADPH in a one-pot reaction, wherein, as a result of at least two further enzymatically catalyzed redox reactions proceeding in the same reaction batch (product-forming reactions), one of the two redox cofactors accumulates in its reduced form and, respectively, the other one in its oxidized form, characterized in that a) in the regeneration reaction which reconverts the reduced cofactor into its original oxidized form, oxygen or a compound of general formula R.sub.1C(O)COOH is reduced, and b) in the regeneration reaction which reconverts the oxidized cofactor into its original reduced form, a compound of general formula R.sub.2CH(OH)R.sub.3 is oxidized and wherein R.sub.1, R.sub.2 and R.sub.3 in the compounds have different meanings.

The present disclosure relates to compositions and methods for producing stereoisomerically pure aminocyclopropanes.

The invention discloses an alcohol dehydrogenase mutant and use thereof. The alcohol dehydrogenase mutant of the present invention has high thermal stability and enables high catalytic efficiency and high conversion rate (i.e. space time yield) in the asymmetric reduction of prochiral diaryl ketones to produce chiral diaryl alcohols. Therefore, the alcohol dehydrogenase mutant of the present invention has extremely high prospect of application in the production of chiral diaryl alcohols, such as (S)-(4-chlorophenyl)-(pyridin-2-yl)-methanol, (R)-(4-chlorophenyl)-(pyridin-2-yl)-methanol.

A monooxygenase having an amino acid sequence obtained by mutation of the amino acid sequence shown in SEQ ID NO:2 is disclosed. The use of the monooxygenase of the present invention in production of chiral sulfoxide-based drugs has advantages including mild reaction conditions, environmental friendliness, high yield, high optical purity of products, less peroxide products, and the like, and therefore the monooxygenase in the present invention has a good industrial application prospect in the production of proton pump inhibitors for the treatment of gastric ulcers.

The present invention provides engineered ketoreductase and phosphite dehydrogenase enzymes having improved properties as compared to a naturally occurring wild-type ketoreductase and phosphite dehydrogenase enzymes, as well as polynucleotides encoding the engineered ketoreductase and phosphite dehydrogenase enzymes, host cells capable of expressing the engineered ketoreductase and phosphite dehydrogenase enzymes, and methods of using the engineered ketoreductase and phosphite dehydrogenase enzymes to synthesize a chiral catalyst used in the synthesis of antiviral compounds, such as nucleoside inhibitors. The present invention further provides methods of using the engineered enzymes to deracemize a chiral alcohol in a one-pot, multi-enzyme system.

An object of the present invention is to provide a method of industrially producing a high-purity L-cyclic amino acid more inexpensively and with a high efficiency, from a cyclic amino acid having a double bond at the 1-position. The present invention provides a method in which an L-cyclic amino acid is produced by allowing a cyclic amino acid having a double bond at the 1-position to react with a specific enzyme having a catalytic ability to reduce a cyclic amino acid having a double bond at the 1-position to produce an L-cyclic amino acid.

A method for producing a compound represented by formula (3) including bringing a carbon-carbon double bond reductase, a microorganism or cell having an ability to produce the enzyme, a processed product of the microorganism or cell, and/or a culture solution containing the enzyme which is obtained by culturing the microorganism or cell, and a carbonyl reductase, a microorganism or cell having an ability to produce the enzyme, a processed product of the microorganism or cell, and/or a culture solution containing the enzyme which is obtained by culturing the microorganism or cell into contact with a compound represented by formula (1) to obtain a compound represented by formula (3): ##STR00001##