The present disclosure relates to a multi-enzyme conjugate, a method for preparing the same and a method for preparing an organic compound using the same. More particularly, a multi-enzyme conjugate exhibiting improved catalytic efficiency over respective free enzymes using site-specific incorporation of a clickable non-natural amino acid into the enzymes and two compatible click reactions, a method for preparing the same and a method for preparing an organic compound using the same may be provided.

Disclosed in the present invention is an L-glutamate dehydrogenase mutant, the sequence of the L-glutamate dehydrogenase mutant being a sequence in which amino acid residue A at position 175 in SEQ ID NO: 1 is mutated to be G, and amino acid residue V at position 386 is mutated to be an amino acid residue having less steric hindrance. Further disclosed in the present invention is an application of the described L-amino acid dehydrogenase mutant in the preparation of L-glufosinate-ammonium or a salt thereof. When the L-glutamate dehydrogenase mutant of the present invention is used to prepare L-glufosinate-ammonium or a salt thereof, compared to an L-glutamate dehydrogenase mutant in which only position 175 or 386 is mutated, the specific enzyme activity is higher. Therefore, the action efficiency of the enzyme is improved, reaction costs are reduced, and industrial production is facilitated.

The present disclosure provides recombinant bacteria for production of indole-3-acetic acid (IAA). Pharmaceutical compositions and methods of treating diseases are also included.

A process comprising (i) providing a gaseous stream including greater than 1% by volume carbon dioxide; (ii) providing water; (iii) converting the carbon dioxide and the water to an organic intermediate and oxygen gas in the presence of light; (iv) separating the oxygen gas from the organic intermediate; and (v) converting the organic intermediate to ethylene and carbon dioxide after said step of separating the oxygen gas from the organic intermediate.

The present invention concerns a method for the production of derivatives of 4-hydroxy-2-ketobutyrate chosen among 1,3-propanediol or 2,4-dihydroxybutyrate by culturing a genetically modified microorganism for the production of the desired derivative of 4-hydroxy-2-ketobutyrate, the microorganism further comprising a gene coding for a mutant glutamate dehydrogenase converting by deamination L-homoserine into 4-hydroxy-2-ketobutyrate. The invention also concerns said genetically modified microorganism.

The present disclosure provides recombinant bacterial cells that have been engineered with genetic circuitry which allow the recombinant bacterial cells to sense a patient's internal environment and respond by turning an engineered metabolic pathway on or off. When turned on, the recombinant bacterial cells complete all of the steps in a metabolic pathway to achieve a therapeutic effect in a host subject. These recombinant bacterial cells are designed to drive therapeutic effects throughout the body of a host from a point of origin of the microbiome. Specifically, the present disclosure provides recombinant bacterial cells comprising a heterologous gene encoding a branched chain amino acid catabolism enzyme. The disclosure further provides pharmaceutical compositions comprising the recombinant bacteria, and methods for treating disorders involving the catabolism of branched chain amino acids using the pharmaceutical compositions disclosed herein.

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.

The invention relates to a whole-cell catalyst which expresses a recombinant α-dioxygenase or the combination of a recombinant fatty acid reductase and a phosphopantetheinyl transferase which phosphopantetheinylates the fatty acid reductase, and which expresses, in addition to the α-dioxygenase and/or the combination of fatty acid reductase and phosphopantetheinyl transferase, a transaminase, wherein the phosphopantetheinyl transferase and/or transaminase is preferably recombinant; and also to a process for converting a carboxylic acid or dicarboxylic acid or a monoester thereof to an amine or diamine, comprising the steps of contacting the carboxylic acid or dicarboxylic acid or the monoester thereof with a phosphopantetheinylated fatty acid reductase or an α-dioxygenase and contacting the product with a transaminase.

The present disclosure provides compositions and methods for rapid production of chemicals in genetically engineered microorganisms in a large scale. Also provided herein is a high-throughput metabolic engineering platform enabling the rapid optimization of microbial production strains. The platform, which bridges a gap between current in vivo and in vitro bio-production approaches, relies on dynamic minimization of the active metabolic network.

The invention provides a whole cell catalyst which expresses a recombinant α-dioxygenase or the combination of a recombinant fatty acid reductase and a phosphopantetheinyl transferase phosphopantetheinylating the fatty acid reductase, and which in addition to the α-dioxygenase and/or the combination of fatty acid reductase and phosphopantetheinyl transferase expresses a transaminase, characterized in that the phosphopantetheinyl transferase and/or transaminase is preferably recombinant; and a method for the conversion of a fatty acid, ω-hydroxy fatty acid, ω-oxo fatty acid or a monoester thereof to an amine, comprising oxidation of the fatty acid, ω-hydroxy fatty acid, ω-oxo fatty acid or the monoester thereof to an oxidation product by contacting with an alkane hydroxylase and/or alcohol dehydrogenase, contacting the oxidation product with a phosphopantetheinylated fatty acid reductase or a α-dioxygenase to give an aldehyde, and contacting the aldehyde with a transaminase.