This disclosure is in the technical field of synthetic biology and metabolic engineering. More particularly, this disclosure is in the technical field of cultivation or fermentation of metabolically engineered cells. This disclosure provides a method for the production of a mixture of at least two different oligosaccharides by a cell as well as the purification of at least one of the oligosaccharides from the cultivation. In addition, this disclosure provides a method for the production of a mixture of at least two different oligosaccharides by a metabolically engineered cell as well as the purification of at least one of the oligosaccharides from the cultivation.

This disclosure is in the technical field of synthetic biology and metabolic engineering. More particularly, this disclosure is in the technical field of cultivation or fermentation of metabolically engineered cells. This disclosure provides a method for the production of a mixture of at least two different oligosaccharides by a cell as well as the purification of at least one of the oligosaccharides from the cultivation. In addition, this disclosure provides a method for the production of a mixture of at least two different oligosaccharides by a metabolically engineered cell as well as the purification of at least one of the oligosaccharides from the cultivation.

The disclosure is in the technical field of synthetic biology and metabolic engineering. More particularly, the disclosure is in the technical field of cultivation or fermentation of metabolically engineered cells. The disclosure describes a cell metabolically engineered for production of an alpha-1,3 glycosylated form of fucose-alpha1,2-galactose-R (Fuc-a1,2-Gal-R). Furthermore, the disclosure provides a method for the production of an alpha-1,3 glycosylated form of Fuc-a1,2-Gal-R by a cell as well as the purification of the alpha-1,3 glycosylated form Fuc-a1,2-Gal-R from the cultivation.

A protein having a novel saccharide oxidase activity capable of being subjected to various uses is provided. The present invention provides a protein having the following physicochemical characteristics: (1) effect: oxidizing a saccharide to produce a saccharic acid; (2) substrate specificity: acting on glucose, maltotriose, maltose, galactose, maltotetraose, lactose, and cellobiose; and, (3) [Km value of glucose]/[Km value of maltose]≤1.

A protein having a novel saccharide oxidase activity capable of being subjected to various uses is provided. The present invention provides a protein having the following physicochemical characteristics: (1) effect: oxidizing a saccharide to produce a saccharic acid; (2) substrate specificity: acting on glucose, maltotriose, maltose, galactose, maltotetraose, lactose, and cellobiose; and, (3) [Km value of glucose]/[Km value of maltose]≤1.

A fermentation product manufacturing process includes fermenting under fermentation conditions in an aqueous fermentation medium in a fermentation reactor a carbohydrate source with a microorganism capable of converting the carbohydrate into a fermentation product which is a salt or a product with a boiling point above the boiling point of water, during the process withdrawing part of the medium including biomass from the reactor in the form of a recycle stream, providing the stream including biomass to a pressure vessel wherein the pressure is such that the temperature of the stream decreases 1-8° C., as compared to the temperature of the medium in the reactor, by the evaporation of water, and recycling the cooled recycle stream to the reactor. The process makes it possible to obtain a homogeneous temperature profile of the fermentation medium with limited occurrence of hot or cool spots within the reactor which results in improved fermentation performance.

The present application discloses a method of culturing a Euglena sp. microorganism, Schizochytrium sp. microorganism, or a Chlorella sp. microorganism comprising culturing the microorganism in a hybrid culture media; maintaining the microorganism heterotrophically in an environment substantially free from light; wherein the hybrid culture media comprises a carbohydrate; and wherein the hybrid culture media comprises fresh media and recycled culture media. The present application also discloses a hybrid culture media.

The present application discloses a method of culturing a Euglena sp. microorganism, Schizochytrium sp. microorganism, or a Chlorella sp. microorganism comprising culturing the microorganism in a hybrid culture media; maintaining the microorganism heterotrophically in an environment substantially free from light; wherein the hybrid culture media comprises a carbohydrate; and wherein the hybrid culture media comprises fresh media and recycled culture media. The present application also discloses a hybrid culture media.

The present invention provides a D-glucaric acid-producing bacterium and a method for producing D-glucaric acid. The present invention is characterized in that D-glucaric acid or a salt thereof is produced from one or more saccharides selected from the group consisting of D-glucose, D-gluconic acid and D-glucuronic acid with catalytic action of a specific alcohol dehydrogenase PQQ-ADH (1) and a specific aldehyde dehydrogenase PQQ-ALDH (2), and that D-glucaric acid or a salt thereof is produced by using a microorganism having the PQQ-ADH (1) and the PQQ-ALDH (2) or a processed product thereof in the presence of the one or more saccharides. The present invention can provide a microorganism having improved productivity of D-glucaric acid to be used for production of D-glucaric acid and a method for efficiently producing D-glucaric acid.

The present invention provides a D-glucaric acid-producing bacterium and a method for producing D-glucaric acid. The present invention is characterized in that D-glucaric acid or a salt thereof is produced from one or more saccharides selected from the group consisting of D-glucose, D-gluconic acid and D-glucuronic acid with catalytic action of a specific alcohol dehydrogenase PQQ-ADH (1) and a specific aldehyde dehydrogenase PQQ-ALDH (2), and that D-glucaric acid or a salt thereof is produced by using a microorganism having the PQQ-ADH (1) and the PQQ-ALDH (2) or a processed product thereof in the presence of the one or more saccharides. The present invention can provide a microorganism having improved productivity of D-glucaric acid to be used for production of D-glucaric acid and a method for efficiently producing D-glucaric acid.