Disclosed herein relates to biopharmaceuticals, and more particularly to a continuous flow method for preparing (R)-3-hydroxy-5-hexenoate. Carbonyl reductase and isopropanol dehydrogenase are co-immobilized onto an inert solid medium simultaneously to prepare a carbonyl reductase/isopropanol dehydrogenase co-immobilized catalyst, which is then filled into a microchannel reactor of the micro reaction system. A solution containing substrate 3-carbonyl-5-hexenoate is subsequently pumped into the microchannel reactor to perform an asymmetric carbonyl reduction reaction to obtain (R)-3-hydroxy-5-hexenoate.

Disclosed are an expression cassette for isopropanol production, a recombinant vector for isopropanol production including the expression cassette, a recombinant microorganism for isopropanol production into which the vector is introduced, and a method of producing isopropanol using the recombinant microorganism. The recombinant microorganism in which a succinic acid bypass metabolic pathway is introduced to an isopropanol production pathway has very high ability to produce isopropanol. The recombinant microorganism is capable of producing isopropanol in an amount corresponding to about 100 times the maximum amount of isopropanol that is produced using known Corynebacterium glutamicum, and thus can effectively produce isopropanol and can be useful in various industrial fields where isopropanol is utilized. The use of the recombinant microorganism makes possible eco-friendly production of high-value-added isopropanol materials for manufacturing biomass-derived chemical products using glucose in lieu of petroleum.

Described is a method for the production isoamyl alcohol (3-methylbutan-1-ol) comprising the enzymatic conversion of 3-methylbutyryl-CoA (isovaleryl-CoA) into isoamyl alcohol comprising: (a) two enzymatic steps comprising (i) first the enzymatic conversion of 3-methylbutyryl-CoA into 3-methylbutyraldehyde (3-methylbutanal or isovaleraldehyde); and (ii) then enzymatically converting the thus obtained 3-methylbutyraldehyde into said isoamyl alcohol; or (b) a single enzymatic reaction in which 3-methylbutyryl-CoA is directly converted into isoamyl alcohol by making use of an alcohol-forming short chain acyl-CoA dehydrogenase/fatty acyl-CoA reductase or an alcohol-forming fatty acyl-CoA reductase (long-chain acyl-CoA:NADPH reductase) (EC 1.2.1.84). Further, described is the above method wherein the 3-methylbutyryl-CoA can be provided by the enzymatic conversion of 3-methylcrotonyl-CoA into said 3-methylbutyryl-CoA. It is also described that the thus obtained isoamyl alcohol can be further enzymatically converted into 3-methylbutyl acetate (isoamyl acetate) as described herein. Described are also recombinant organisms or microorganisms which are capable of performing the above enzymatic conversions. Furthermore, described are uses of enzymes and enzyme combinations which allow the above enzymatic conversions.

The present invention relates to thermophilic cells and methods for the microbial production of volatile compounds, including acetone, butanone and isopropanol. Also provided are nucleic acid constructs, vectors and host cells useful in such methods.

Described is a method for the production isoamyl alcohol (3-methylbutan-1-ol) comprising the enzymatic conversion of 3-methylbutyryl-CoA (isovaleryl-CoA) into isoamyl alcohol comprising: (a) two enzymatic steps comprising (i) first the enzymatic conversion of 3-methylbutyryl-CoA into 3-methylbutyraldehyde (3-methylbutanal or isovaleraldehyde); and (ii) then enzymatically converting the thus obtained 3-methylbutyraldehyde into said isoamyl alcohol; or (b) a single enzymatic reaction in which 3-methylbutyryl-CoA is directly converted into isoamyl alcohol by making use of an alcohol-forming short chain acyl-CoA dehydrogenase/fatty acyl-CoA reductase or an alcohol-forming fatty acyl-CoA reductase (long-chain acyl-CoA:NADPH reductase) (EC 1.2.1.84). Further, described is the above method wherein the 3-methylbutyryl-CoA can be provided by the enzymatic conversion of 3-methylcrotonyl-CoA into said 3-methylbutyryl-CoA. It is also described that the thus obtained isoamyl alcohol can be further enzymatically converted into 3-methylbutyl acetate (isoamyl acetate) as described herein. Described are also recombinant organisms or microorganisms which are capable of performing the above enzymatic conversions. Furthermore, described are uses of enzymes and enzyme combinations which allow the above enzymatic conversions.

The present disclosure relates to the technical field of biochemical engineering and particularly discloses a preparation method for (R)-3-hydroxyl-5-hexenoate. In the method of the present disclosure, the (R)-3-hydroxyl-5-hexenoate is prepared by catalytic reduction of 3-carbonyl-5-hexenoate by ketoreductase with 3-carbonyl-5-hexenoate as the substrate. The amino acid sequence of ketoreductase is shown in SEQ ID NO.1. In the present disclosure, the (R)-3-hydroxyl-5-hexenoate having a very high chiral purity is obtained by asymmetric reduction by ketoreductase as the biocatalyst. The present disclosure has the advantages of easy operation, mild reaction conditions, high reaction yield and good practical industrial application value.

Disclosed herein relates to biopharmaceuticals, and more particularly to a continuous flow method for preparing (R)-3-hydroxy-5-hexenoate. Carbonyl reductase and isopropanol dehydrogenase are co-immobilized onto an inert solid medium simultaneously to prepare a carbonyl reductase/isopropanol dehydrogenase co-immobilized catalyst, which is then filled into a microchannel reactor of the micro reaction system. A solution containing substrate 3-carbonyl-5-hexenoate is subsequently pumped into the microchannel reactor to perform an asymmetric carbonyl reduction reaction to obtain (R)-3-hydroxy-5-hexenoate.

This document describes biochemical pathways for producing 1,3-butanediol using a polypetide having -ketothiolase activity to form a 3-oxo-5-hydroxypentanoyl-CoA intermediate that can be enzymatically converted to 1,3-butanediol, as well as recombinant hosts producing 1,3-butanediol.

Recombinant yeasts bioengineered to overexpress genes for utilization of fatty acids to produce acetone and isopropanol, and methods of use thereof. The yeasts are modified to express, constitutively express, or overexpress an acetyl-CoA thioesterase, an acetyl-CoA C-acetyltransferase, an acetoacetyl-CoA transferase, an acetoacetyl-CoA thioesterase, an acetoacetate decarboxylase, an isopropanol dehydrogenase, or any combination thereof. The methods include cultivating the recombinant yeasts to convert any fatty acid-containing feedstocks into acetone and/or isopropanol.

This document describes biochemical pathways for producing 1,3-butanediol using a polypetide having -ketothiolase activity to form a 3-oxo-5-hydroxypentanoyl-CoA intermediate that can be enzymatically converted to 1,3-butanediol, as well as recombinant hosts producing 1,3-butanediol.