The current disclosure provides a process for enzymatically converting a saccharide into allulose. The invention also relates to a process for preparing allulose where the process involves converting fructose 6-phosphate (F6P) to allulose 6-phosphate (A6P), catalyzed by allulose 6-phosphate 3-epimerase (A6PE), and converting the A6P to allulose, catalyzed by allulose 6-phosphate phosphatase (A6PP).

The invention provides methods for glycosylation and preparation of compounds. The compounds include galactosylated, sialylated, fucosylated, and N-acetylglucosaminylated compounds from simple animal-derived, plant-derived, or microbe-derived oligosaccharides and sugars. In certain embodiments, the invention provides trinucleotide-free enzymatic production of oligosaccharides starting from simple sugars that include plant-based sugars. The invention also provides the enzymatic production of fucosylated oligosaccharides and fucosylated antibody-glycan conjugates from common sugars. The production may be a cell-free, one-pot synthesis using enzymes, and in some embodiments, immobilized enzymes. The synthesis is a highly customizable and highly efficient cell-free manufacturing process. In some embodiments, lactose derivatives and human milk oligosaccharides (HMOs) are produced.

The current disclosure provides a process for enzymatically converting a saccharide into allulose. The invention also relates to a process for preparing allulose where the process involves converting fructose 6-phosphate (F6P) to allulose 6-phosphate (A6P), catalyzed by allulose 6-phosphate 3-epimerase (A6PE), and converting the A6P to allulose, catalyzed by allulose 6-phosphate phosphatase (A6PP).

A method for preparing a yeast capable of simultaneously fermenting a pentose and a hexose sugar, comprising: (a) providing a yeast comprising: one or more heterologous genes encoding an enzyme of a pentose metabolic pathway; disruptions of a gene encoding a ribulose-phosphate 3-epimerase and of a gene encoding a glucose-6-phosphate isomerase; one or more overexpressed endogenous genes encoding an enzyme of the pentose phosphate pathway; and a disruption of one or more genes encoding an NADPH dependent 6-phosphogluconate dehydrogenase, (b) subjecting said yeast to evolutionary engineering on a medium comprising a hexose sugar and at least one pentose sugar, selecting for a yeast with improved growth rate obtain an evolved yeast; (d) restoring, in the evolved yeast, one or more of the disrupted genes, or: (d) identifying genetic permutations in at least part of the genome of the evolved yeast by genome sequencing; (e) constructing an improved pentose and hexose-fermenting yeast comprising one or more said genetic permutations. Also described is a recombinant yeast comprising one or more heterologous genes of a pentose metabolic pathway, and a gene encoding a variant of a parent polypeptide, the variant comprising an amino acid sequence comprising at least one mutation, when aligned with the amino acid sequence in SEQ ID NO: 6.

Provided are a 3-epimerase, an encoding polynucleotide therefor, a nucleic acid construct, vector, and host cell comprising the polynucleotide, a method for producing the 3-epimerase, and use of the 3-epimerase.

The present application relates to recombinant microorganisms useful in the biosynthesis of monoethylene glycol (MEG) or glycolic acid (GA), or MEG and one or more co-product, from one or more pentose and/or hexose sugars. Also provided are methods of producing MEG (or GA), or MEG (or GA) and one or more co-product, from one or more pentose and/or hexose sugars using the recombinant microorganisms, as well as compositions comprising the recombinant microorganisms and/or the products MEG (or GA), or MEG and one or more co-product.

The present application relates to recombinant microorganisms useful in the biosynthesis of monoethylene glycol (MEG), or optionally MEG and one or more co-product, from one or more hexose feedstock. The present application also relates to recombinant microorganisms useful in the biosynthesis of glycolic acid (GA), or optionally GA and one or more co-product, from one or more hexose feedstock. The present application relates to recombinant microorganisms useful in the biosynthesis of xylitol, or optionally xylitol and one or more co-product, from one or more hexose feedstock. Also provided are methods of producing MEG (or GA or xylitol), or optionally MEG (or GA or xylitol) and one or more co-product, from one or more hexose feedstock using the recombinant microorganisms, as well as compositions comprising the recombinant microorganisms and/or the products MEG (or GA or xylitol), or optionally MEG (or GA or xylitol) and one or more co-product.

An acetic acid metabolizing ability of a recombinant yeast strain having xylose-metabolizing ability is to be improved. In such a recombinant yeast strain having xylose-metabolizing ability, the acetaldehyde dehydrogenase gene has been introduced and a gene encoding NADH dehydrogenase involved in reoxidation of cytoplasmic NADH on the mitochondrial outer membrane has been suppressed.

The current disclosure provides a process for enzymatically converting a saccharide into allulose. The invention also relates to a process for preparing allulose where the process involves converting fructose 6-phosphate (F6P) to allulose 6-phosphate (A6P), catalyzed by allulose 6-phosphate 3-epimerase (A6PE), and converting the A6P to allulose, catalyzed by allulose 6-phosphate phosphatase (A6PP).

The current disclosure provides a process for enzymatically converting a saccharide into allulose. The invention also relates to a process for preparing allulose where the process involves converting fructose 6-phosphate (F6P) to allulose 6-phosphate (A6P), catalyzed by allulose 6-phosphate 3-epimerase (A6PE), and converting the A6P to allulose, catalyzed by allulose 6-phosphate phosphatase (A6PP).