The present application discloses a simple process for the purification of neutral human milk oligosaccharides (HMOs) produced by microbial fermentation. The process uses a combination of cationic ion exchanger treatment, an anionic ion exchanger treatment, and a nanofiltration and/or electrodialysis step, which allows efficient purification of large quantities of neutral HMOs at high purity. Contrary to the purification currently used in fermentative production of neutral HMOs, the presented process allows the provision of HMOs without the need of a chromatographic separation. The so purified HMOs may be obtained in solid form by spray drying, as crystalline material or as sterile filtered concentrate. The provided HMOs are free of proteins and recombinant material originating from the used recombinant microbial strains and thus very well-suited for use in food, medical food and feed (e.g. pet food) applications.

The present application discloses a simple process for the purification of neutral human milk oligosaccharides (HMOs) produced by microbial fermentation. The process uses a combination of cationic ion exchanger treatment, an anionic ion exchanger treatment, and a nanofiltration and/or electrodialysis step, which allows efficient purification of large quantities of neutral HMOs at high purity. Contrary to the purification currently used in fermentative production of neutral HMOs, the presented process allows the provision of HMOs without the need of a chromatographic separation. The so purified HMOs may be obtained in solid form by spray drying, as crystalline material or as sterile filtered concentrate. The provided HMOs are free of proteins and recombinant material originating from the used recombinant microbial strains and thus very well-suited for use in food, medical food and feed (e.g. pet food) applications.

An improved dry grind system and method for producing a sugar stream from grains or similar carbohydrate sources and/or residues, such as for biochemical production. In particular, after saccharification and prior to a sugar conversion process, a sugar/carbohydrate stream is removed from a saccharified stream. The sugar/carbohydrate stream includes a desired Dextrose Equivalent (DE) where DE describes the degree of conversion of starch to dextrose can be produced, with the such sugar stream being available for biochemical production, e.g., alcohol production, or other processes. In addition, the systems and methods also can involve the removal of certain grain components, e.g., corn kernel components, including protein and/or fiber. Sugar stream production occurs on the front end of the system and method.

An improved dry grind system and method for producing a sugar stream from grains or similar carbohydrate sources and/or residues, such as for biochemical production. In particular, after saccharification and prior to a sugar conversion process, a sugar/carbohydrate stream is removed from a saccharified stream. The sugar/carbohydrate stream includes a desired Dextrose Equivalent (DE) where DE describes the degree of conversion of starch to dextrose can be produced, with the such sugar stream being available for biochemical production, e.g., alcohol production, or other processes. In addition, the systems and methods also can involve the removal of certain grain components, e.g., corn kernel components, including protein and/or fiber. Sugar stream production occurs on the front end of the system and method.

The present invention can: efficiently and readily produce glucose from microalgae that accumulate starch in their cells; and obtain ethanol. During a preparation step of the glucose production method, microalgae are prepared on which a saccharifying enzyme acts on starch accumulated inside the microalgae cells, without disrupting the cell walls. In a saccharification step, starch inside the cells is saccharified and glucose is generated, by adding a saccharifying enzyme to the microalgae without a disruption treatment. The ethanol production method has a step in which, after the saccharification step, the glucose undergoes alcoholic fermentation and ethanol is generated.

The present invention provides Streptomyces coelicolor strain A3(2)_M22-2C43 obtained by inducing a point mutation in the base sequence of the DagB gene in a wild-type Streptomyces coelicolor strain A3(2) by UV radiation. Since the Streptomyces coelicolor strain A3(2)_M22-2C43 according to the present invention expresses a DagB mutant enzyme expressing little or no DagB beta-agarase or exhibiting little or no beta-agarase activity, there is no need for separate isolation and purification of DagA enzymes from culture fluid, and the culture fluid of the Streptomyces coelicolor strain A3(2)_M22-2C43 or supernatant thereof may be used to produce, from agar or agarose, neoagarose oligosaccharides with a higher content of neoagarotetraose or neoagarohexaose than that of neoagarobiose.

The present invention provides Streptomyces coelicolor strain A3(2)_M22-2C43 obtained by inducing a point mutation in the base sequence of the DagB gene in a wild-type Streptomyces coelicolor strain A3(2) by UV radiation. Since the Streptomyces coelicolor strain A3(2)_M22-2C43 according to the present invention expresses a DagB mutant enzyme expressing little or no DagB beta-agarase or exhibiting little or no beta-agarase activity, there is no need for separate isolation and purification of DagA enzymes from culture fluid, and the culture fluid of the Streptomyces coelicolor strain A3(2)_M22-2C43 or supernatant thereof may be used to produce, from agar or agarose, neoagarose oligosaccharides with a higher content of neoagarotetraose or neoagarohexaose than that of neoagarobiose.

The present disclosure provides methods for generating sugars from a cellulosic biomass. The methods combine treatment of the biomass using a high-shear milling device and saccharification of the biomass to partially hydrolyze the biomass. The biomass can be saccharified either after or simultaneously with the high-shear milling treatment. The partially hydrolyzed biomass is then separated into a solids stream with saccharification enzymes, and a liquid stream with sugars. The solids stream and associated enzymes are further incubated under saccharification conditions to produce additional sugars, or are recycled and added to fresh biomass, which is saccharified under high-shear milling conditions. The methods result in improved conversion of cellulosic biomass to glucose.

The present disclosure provides methods for generating sugars from a cellulosic biomass. The methods combine treatment of the biomass using a high-shear milling device and saccharification of the biomass to partially hydrolyze the biomass. The biomass can be saccharified either after or simultaneously with the high-shear milling treatment. The partially hydrolyzed biomass is then separated into a solids stream with saccharification enzymes, and a liquid stream with sugars. The solids stream and associated enzymes are further incubated under saccharification conditions to produce additional sugars, or are recycled and added to fresh biomass, which is saccharified under high-shear milling conditions. The methods result in improved conversion of cellulosic biomass to glucose.

The present invention is directed to methods and kits for template-free enzymatic synthesis of polynucleotides that include or enable a step of efficiently cleaving the polynucleotide products from its initiator using an endonuclease V activity and initiator with a 3′-penultimate deoxyinosine.