This invention relates to the production of chemicals by fermentation with a microorganism in which the fermentation medium contains the sugar sucrose. As a specific example, succinic acid is produced from a sucrose-containing renewable feedstock through fermentation using a biocatalyst. Examples of such a biocatalyst include microorganisms that have been enhanced in their ability to utilize sucrose as a carbon and energy source. The biocatalysts of the present invention are derived from the genetic manipulation of parental strains that were originally constructed with the goal to produce one or more chemicals (for example succinic acid and/or a salt of succinic acid) at a commercial scale using feedstocks other than sucrose. The genetic manipulations of the present invention involve the introduction of exogenous genes involved in the transport and metabolism of sucrose into the parental strains. The genes involved in the transport and metabolism of sucrose can also be introduced into a microorganism prior to developing the organism to produce a particular chemical. The genes involved in the transport and metabolism of sucrose can also be used to augment or improve the sucrose transport and metabolism by strains already known to have some ability for sucrose utilization in biological fermentation.

This invention relates to the production of chemicals by fermentation with a microorganism in which the fermentation medium contains the sugar sucrose. As a specific example, succinic acid is produced from a sucrose-containing renewable feedstock through fermentation using a biocatalyst. Examples of such a biocatalyst include microorganisms that have been enhanced in their ability to utilize sucrose as a carbon and energy source. The biocatalysts of the present invention are derived from the genetic manipulation of parental strains that were originally constructed with the goal to produce one or more chemicals (for example succinic acid and/or a salt of succinic acid) at a commercial scale using feedstocks other than sucrose. The genetic manipulations of the present invention involve the introduction of exogenous genes involved in the transport and metabolism of sucrose into the parental strains. The genes involved in the transport and metabolism of sucrose can also be introduced into a microorganism prior to developing the organism to produce a particular chemical. The genes involved in the transport and metabolism of sucrose can also be used to augment or improve the sucrose transport and metabolism by strains already known to have some ability for sucrose utilization in biological fermentation.

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.

An acetyl-CoA producing microorganism obtained by imparting at least one enzymatic activity selected from the group consisting of malate thiokinase, malyl-CoA lyase, glyoxylate carboligase, 2-hydroxy-3-oxopropionate reductase, and hydroxypyruvate reductase, to a microorganism that does not have any of the following (a), (b), (c), (d) or (e): (a) a carbon dioxide fixation cycle including an enzymatic reaction from malonyl-CoA to malonate semialdehyde or 3-hydroxypropionate; (b) a carbon dioxide fixation cycle including an enzymatic reaction from acetyl-CoA and CO.sub.2 to pyruvate; (c) a carbon dioxide fixation cycle including an enzymatic reaction from crotonyl-CoA and CO.sub.2 to ethylmalonyl-CoA or glutaconyl-CoA; (d) a carbon dioxide fixation cycle including an enzymatic reaction from CO.sub.2 to formate; or (e) at least one selected from the group consisting of malate thiokinase and malyl-CoA lyase.

An acetyl-CoA producing microorganism obtained by imparting at least one enzymatic activity selected from the group consisting of malate thiokinase, malyl-CoA lyase, glyoxylate carboligase, 2-hydroxy-3-oxopropionate reductase, and hydroxypyruvate reductase, to a microorganism that does not have any of the following (a), (b), (c), (d) or (e): (a) a carbon dioxide fixation cycle including an enzymatic reaction from malonyl-CoA to malonate semialdehyde or 3-hydroxypropionate; (b) a carbon dioxide fixation cycle including an enzymatic reaction from acetyl-CoA and CO.sub.2 to pyruvate; (c) a carbon dioxide fixation cycle including an enzymatic reaction from crotonyl-CoA and CO.sub.2 to ethylmalonyl-CoA or glutaconyl-CoA; (d) a carbon dioxide fixation cycle including an enzymatic reaction from CO.sub.2 to formate; or (e) at least one selected from the group consisting of malate thiokinase and malyl-CoA lyase.

The present invention relates to KatG enzymes and enzyme compositions and their uses in enzymatic degradation of plastics.

The disclosure discloses an Aspergillus niger engineered strain for reducing byproduct succinic acid in a fermentation process of L-malic acid. The Aspergillus niger engineered strain is an Aspergillus niger engineered strain in which fumaric acid reductase frdA and fumaric acid reductase flavoprotein subunit frdB are simultaneously knocked out. The disclosure provides an frdA and frdB gene double-knockout Aspergillus niger strain, and a method for greatly reducing byproduct succinic acid in a fermentation process of L-malic acid. By the disclosure, the byproduct succinic acid accumulated in a production process of malic acid through fermentation of Aspergillus niger is significantly reduced, a cost in a downstream separation and purification process of malic acid is decreased, and good strains are provided for producing malic acid via industrial fermentation.

The disclosure discloses an Aspergillus niger engineered strain for reducing byproduct succinic acid in a fermentation process of L-malic acid. The Aspergillus niger engineered strain is an Aspergillus niger engineered strain in which fumaric acid reductase frdA and fumaric acid reductase flavoprotein subunit frdB are simultaneously knocked out. The disclosure provides an frdA and frdB gene double-knockout Aspergillus niger strain, and a method for greatly reducing byproduct succinic acid in a fermentation process of L-malic acid. By the disclosure, the byproduct succinic acid accumulated in a production process of malic acid through fermentation of Aspergillus niger is significantly reduced, a cost in a downstream separation and purification process of malic acid is decreased, and good strains are provided for producing malic acid via industrial fermentation.

The disclosure discloses an Aspergillus niger engineered strain for reducing byproduct fumaric acid in a fermentation process of L-malic acid. The Aspergillus niger engineered strain is an Aspergillus niger engineered strain in which a fumarate hydratase gene fum is knocked out. The disclosure overcomes the defects in the prior art, in the current process of producing malic acid through fermentation of Aspergillus niger, byproduct fumaric acid can be accumulated with the generation of malic acid so as to cause the improved cost of the subsequent malic acid purification process. The disclosure provides an Aspergillus niger engineered strain in which a fum gene is knocked out and a method for greatly reducing byproduct fumaric acid in the fermentation production of Aspergillus niger.

The disclosure discloses an Aspergillus niger engineered strain for reducing byproduct fumaric acid in a fermentation process of L-malic acid. The Aspergillus niger engineered strain is an Aspergillus niger engineered strain in which a fumarate hydratase gene fum is knocked out. The disclosure overcomes the defects in the prior art, in the current process of producing malic acid through fermentation of Aspergillus niger, byproduct fumaric acid can be accumulated with the generation of malic acid so as to cause the improved cost of the subsequent malic acid purification process. The disclosure provides an Aspergillus niger engineered strain in which a fum gene is knocked out and a method for greatly reducing byproduct fumaric acid in the fermentation production of Aspergillus niger.