Glycan polymers and related methods thereof
AssigneeKaleido Biosciences, Inc.
InventorsMolly Krisann Gibson, Christopher Matthew Liu, Geoffrey A. von Maltzahn, Han Yuan
Life of this patent
Estimating the term…
Abstract
Compositions of glycan polymers and methods of making and manufacturing the same are described herein. Also provided are methods of treating a disease or disorder with a glycan polymer preparation.
Claims460
A method of treating a subject having a disease or disorder associated with an unwanted level of a metabolite (e.g., a short chain fatty acid (SCFA) (e.g., propionate or butyrate), ammonia, trimethylamine (TMA), trimethylamine N-oxide (TMAO), a uremic solute (e.g., p-cresol or indole), lipopolysaccharide (LPS), or a bile acid (e.g., a secondary bile acid)), comprising:
- optionally, selecting a glycan polymer preparation on the basis that it modulates the production or level of the metabolite, and
- administering an amount of the glycan polymer preparation effective to result in a modulation of the level of the metabolite, thereby treating the disease or disorder.
A method of treating a subject having a disease or disorder associated with an unwanted level of a metabolite (e.g., a short chain fatty acid (SCFA) (e.g., propionate or butyrate), ammonia, trimethylamine (TMA), trimethylamine N-oxide (TMAO), a uremic solute (e.g., p-cresol or indole), lipopolysaccharide (LPS), or a bile acid (e.g., a secondary bile acid)), comprising:
- optionally, acquiring knowledge that a glycan polymer preparation modulates the production or level of the metabolite, and
- administering an amount of the glycan polymer preparation effective to result in a modulation of the level of the metabolite, thereby treating the disease or disorder.
3A. The method of any of claims 1-3, wherein the glycan polymers, or at least 20, 30, 40, 50, 60, 70, 80, 90, 95, or 99% (by weight or number) of the glycan polymers, of the glycan polymer preparation have one or more (e.g. two, three, four, five, or six) of the properties listed in Table 1, optionally selected from:
- a. glycan polymers comprising a glucose, mannose, or galactose subunit, or a combination thereof and at least one alpha-glycosidic bond,
- b. glycan polymers comprising a glucose, mannose, or galactose subunit, or a combination thereof and at least one beta-glycosidic bond,
- c. glycan polymers comprising a xylose, arabinose, fucose or rhamnose subunit, or a combination thereof and at least one alpha-glycosidic bond,
- d. glycan polymers comprising a xylose, arabinose, fucose or rhamnose subunit, or a combination thereof and at least one beta-glycosidic bond,
- e. glycan polymers comprising a glucose or galactose subunit, or a combination thereof and at least one alpha-glycosidic bond, or
- f. glycan polymers comprising a glucose or galactose subunit, or a combination thereof and at least one beta-glycosidic bond.
A method of modulating the production or level of a product (e.g., a short chain fatty acid (SCFA), ammonia, trimethylamine (TMA), trimethylamine N-oxide (TMAO), a uremic solute, or a bile acid) in the body (e.g., the gut (colon, intestine), blood, urine, an organ (e.g. liver, kidney), the brain) of a subject comprising:
- administering (e.g. orally or rectally) an effective amount of a glycan polymer preparation to the subject sufficient to modulate the production or level of a product, optionally, wherein the glycan polymer is a substrate for a microbial constituent of the colon or intestine.
A method of selecting a glycan polymer preparation for use as a substrate for a glycosidase enzyme (e.g. CAZy family) of a preselected human gut microbe (e.g. selected because of its glycosidase profile), comprising:
- a) acquiring a value for the glycosidase (e.g. CAZy family) profile of a microbe,
- b) identifying, designing, or selecting a glycan polymer capable of being a substrate of the microbe on the basis of the glycosidase (e.g. CAZy family) profile,
- c) optionally,
- i. assembling a panel of human gut microbes (e.g. single strains, designed communities of strains, or ex vivo communities, e.g. from fecal samples, which include the microbe of interest)
- ii. contacting the panel of microbes with a test glycan preparation,
- iii. assessing the growth of the human gut microbe (of interest)
- d) selecting the glycan polymer preparation.
A glycan polymer preparation comprising glycan polymers, e.g., wherein the preparation comprises at least 0.5, 1, 2, 5, 10, 50, or 100 kg, and, e.g., is at least 20, 30, 40, 50, 60, 70, 80, 90, 95 or 99% pure, comprising:
- i) a glucose, mannose, or galactose subunit, or a combination thereof and at least one alpha-glycosidic bond, or
- ii) a glucose, mannose, or galactose subunit, or a combination thereof and at least one beta-glycosidic bond, and
- which are a substrate of one or more, e.g., two, three, four, or more, human gut microbe glycosidase enzymes selected from:
- i) GT5, GH94, GH13 subfamily 9, GH13 subfamily 39, GH13 subfamily 36, GH113 or GH112 CAZy family,
- ii) GT2, GT4, GT5, GT35, GT51, GH1, GH2, GH3, GH4, GH13, GH13 subfamily 9, GH13 subfamily 31, GH18, GH23, GH25, GH28, GH31, GH32, GH36, GH51, GH73, GH77, or GH94 CAZy family,
- iii) GT11, GT10, GH92, GH51, GH35, GH29, GH28, GH20, GH130, GH13 subfamily 8, or GH13 subfamily 14 CAZy family, or
- iv) GT2, GT4, GH2, GH23, GH3, GT8, GT51, GT9, GH1, GH92, GH73, GH31, GH20, GH28, GT25, GT28, GT35, GH18, GT0, GH13, GH36, GH97, GH105, GH25, GH4, GH32, GH78, GH29, GH0, GH51, GT10, or GH77 CAZy family.
A glycan polymer preparation, e.g., wherein the preparation comprises at least about 0.5, 1, 2, 5, 10, 50, or 100 kg, and, e.g., is at least 20, 30, 40, 50, 60, 70, 80, 90, 95 or 99% pure, comprising glycan polymers comprising:
- i) a xylose, arabinose, fucose or rhamnose subunit, or a combination thereof and at least one alpha-glycosidic bond, or
- ii) a xylose, arabinose, fucose or rhamnose subunit, or a combination thereof and at least one beta-glycosidic bond, and
- which are a substrate of one or more, e.g., two, three, four, or more, human gut microbe glycosidase enzymes selected from:
- i) GT11, GT10, GH92, GH51, GH35, GH29, GH28, GH20, GH130, GH13 subfamily 8, or GH13 subfamily 14 CAZy family, or
- ii) GT2, GT4, GH2, GH23, GH3, GT8, GT51, GT9, GH1, GH92, GH73, GH31, GH20, GH28, GT25, GT28, GT35, GH18, GT0, GH13, GH36, GH97, GH105, GH25, GH4, GH32, GH78, GH29, GH0, GH51, GT10, or GH77 CAZy family.
A glycan polymer preparation, e.g., wherein the preparation comprises at least 0.5, 1, 2, 5, 10, 50, or 100 kg, and, e.g., is at least 20, 30, 40, 50, 60, 70, 80, 90, 95 or 99% pure, comprising glycan polymers comprising:
- i) a glucose or galactose subunit, or a combination thereof and at least one alpha-glycosidic bond, or
- ii) a glucose or galactose subunit, or a combination thereof and at least one beta-glycosidic bond, and
- which are a substrate of one or more, e.g., two, three, four, or more, human gut microbe glycosidase enzymes selected from:
- i) GT3, GH97, GH43 subfamily 24, GH27, GH133, GH13 subfamily 8, GH13 CAZy family, or
- ii) GT2, GT4, GH2, GH23, GH3, GT8, GT51, GT9, GH1, GH92, GH73, GH31, GH20, GH28, GT25, GT28, GT35, GH18, GT0, GH13, GH36, GH97, GH105, GH25, GH4, GH32, GH78, GH29, GH0, GH51, GT10, GH77, GT2, GT4, GH2, GH23, GH3, GT51, GH1, GT8, GH92, GT9, GH73, GH31, GH20, Gh28, GT35, GT28, GH18, GH13, GH97, GH25, GH36, GH4, GH105, GH32, GH78, GH29, GH0, GT25, GH51, GH77, GH88, GH24 CAZy family.
A glycan polymer preparation, e.g., wherein the preparation comprises at least 0.5, 1, 2, 5, 10, 50, or 100 kg, and, e.g., is at least 20, 30, 40, 50, 60, 70, 80, 90, 95 or 99% pure, comprising glycan polymers comprising:
- an arabinose, galactose, xylose, or glucose subunit, or a combination thereof and at least one alpha-glycosidic bond, and
- which are a substrate of one or more, e.g., two, three, four, or more, human gut microbe glycosidase enzymes selected from:
- i) GH13 subfamily 3, GH13 subfamily 30, GH30 subfamily 2, GH30 subfamily 5, GH43 subfamily 22, GH43 subfamily 8, or GH84 CAZy family, or
- ii) GH3, GH106, GH105, GH2, GH20, GH28, GH76, GH97, or GH92 CAZy family.
A glycan polymer preparation, e.g., wherein the preparation comprises at least 0.5, 1, 2, 5, 10, 50, or 100 kg, and, e.g., is at least 20, 30, 40, 50, 60, 70, 80, 90, 95 or 99% pure, comprising glycan polymers comprising:
- a glucose and at least one alpha-glycosidic bond, and
- which are a substrate of one or more, e.g., two, three, four, or more, human gut microbe glycosidase enzymes selected from:
- i) GH13 subfamily 19, GH13 subfamily 21, GH23, GH33, GH37 or GH104 CAZy family, or
- ii) GH23, GH24, or GH33 CAZy family.
A glycan polymer preparation, e.g., wherein the preparation comprises at least 0.5, 1, 2, 5, 10, 50, or 100 kg, and, e.g., is at least 20, 30, 40, 50, 60, 70, 80, 90, 95 or 99% pure, comprising glycan polymers comprising:
- i) a glucose or xylose subunit, or a combination thereof and at least one alpha-glycosidic bond, or
- ii) a glucose or xylose subunit, or a combination thereof and at least one beta-glycosidic bond, and
- which are a substrate of one or more, e.g., two, three, four, or more, human gut microbe glycosidase enzymes selected from:
- i) GH13 subfamily 20, GH13 subfamily 31, GH13 subfamily 39, GH39, GH43 subfamily 11, GH5 subfamily 44, or GH94 CAZy family, or
- ii) GH2, GH31, GH23, GH13, or GH24 CAZy family.
A glycan polymer preparation, e.g., wherein the preparation comprises at least 0.5, 1, 2, 5, 10, 50, or 100 kg, and, e.g., is at least 20, 30, 40, 50, 60, 70, 80, 90, 95 or 99% pure, comprising glycan polymers comprising:
- a glucose, xylose, arabinose, or galactose subunit, or a combination thereof and at least one alpha-glycosidic bond, and
- which are a substrate of one or more, e.g., two, three, four, or more, human gut microbe glycosidase enzymes selected from:
- i) GH13 subfamily 3, GH13 subfamily 30, GH121, GH15, GH43 subfamily 27, GH43 subfamily 34, or GH43 subfamily 8 CAZy family, or
- ii) GH92, GH97, GH76, GH28, GH20, GH105, GH2, GH50, GH3, or GH106 CAZy family.
A method of treating a subject having a dysbiosis, comprising:
- administering a composition comprising a glycan polymer preparation described herein and a preparation of a microbe in an amount effective to treat the dysbiosis.
A glycan polymer preparation described herein comprising glycan polymers which are a substrate of a human gut microbe glycosidase enzyme of a spore-forming microbe (e.g. spore-forming bacterial taxa)
A glycan polymer preparation, optionally, e.g., wherein the preparation comprises at least about 0.5, 1, 2, 5, 10, 50, or 100 kg, and/or, further optionally, e.g., is at least 20, 30, 40, 50, 60, 70, 80, 90, 95 or 99% pure, comprising glycan polymers comprising:
- a. a xylose or arabinose subunit, or a combination thereof and at least one alpha-glycosidic bond,
- b. a xylose or arabinose subunit, or a combination thereof and at least one beta-glycosidic bond,
- c. a galactose, xylose, or arabinose subunit, or a combination thereof and at least one alpha-glycosidic bond,
- d. a galactose, xylose, or arabinose subunit, or a combination thereof and at least one beta-glycosidic bond,
- e. a glucose, xylose, or arabinose subunit, or a combination thereof and at least one alpha-glycosidic bond,
- f. a glucose, xylose, or arabinose subunit, or a combination thereof and at least one beta-glycosidic bond,
- g. a xylose, arabinose, glucose or galactose subunit, or a combination thereof and at least one alpha-glycosidic bond,
- h. a xylose, arabinose, glucose or galactose subunit, or a combination thereof and at least one beta-glycosidic bond, or a combination thereof and at least one beta-glycosidic bond, and
- which are a substrate of a human gut microbe glycosidase enzyme of one of:
- GT5, GT35, GT3, GH97, GH95, GH92, GH89, GH88, GH78, GH77, GH57, GH51, GH43 subfamily 34, GH43 subfamily 24, GH43 subfamily 10, GH42, GH36, GH35, GH33, GH32, GH31, GH3, GH29, GH28, GH27, GH24, GH20, GH2, GH16, GH133, GH130, GH13 subfamily 8, GH13 subfamily 38, GH13 subfamily 14, GH13, GH123, GH115, GH109, or GH105 CAZy family.
A method of making a co-preparation comprising:
- providing a preparation of a spore-forming microbe (e.g. a spore-forming human gut microbe),
- providing the glycan polymer preparation (described herein),
- wherein the glycan polymer is a substrate of the spore-forming microbe, and
- combining the preparation of the spore-forming microbe with the glycan polymer preparation.
A method of any embodiment described herein.
A composition of any embodiment described herein.
A method of making a preparation of a glycan polymer, e.g., a glycan polymer that is a substrate for a glycosidase enzyme present in a human gut microbe, comprising:
- providing a plurality of glycan subunits, e.g., a sugar monomer or a sugar dimer, suitable for the production of the glycan polymer;
- and
- contacting the glycan subunits of the plurality with a glycosidase enzyme molecule, e.g. derived from a human gut microbe, under conditions that result in the incorporation, e.g., by a condensation reaction, of the glycan subunits into a glycan polymer,
- thereby making a glycan polymer preparation that is a substrate for a human gut microbe, optionally wherein:
- i) the glycan polymer preparation comprises at least about 0.25, 0.5, 1, 5, 10, 20, 50, 100, 200, 300, 400 or 500 kilograms of glycan polymer, and/or
- ii) the glycan polymer preparation is produced at a yield of at least about 15%, 30%, 45%, 60%, or of about 75% (as determined on a weight/weight basis as a percentage of input glycan subunits).
A method of making a glycan polymer preparation, comprising:
- providing a plurality of glucose, mannose, and/or galactose containing glycan subunits (e.g., monomers or dimers);
- contacting the plurality of glycan subunits with a glycosidase enzyme selected from one of GT5, GH94, GH13 subfamily 9, GH13 subfamily 39, GH13 subfamily 36, GH113 or GH112 CAZy family;
- under conditions that result in making a glycan polymer preparation, wherein a glycan polymer of the preparation is a substrate for a human gut microbe comprising a glycosidase enzyme of a GT5, GH94, GH13 subfamily 9, GH13 subfamily 39, GH13 subfamily 36, GH113 or GH112 CAZy family.
A method of making a glycan polymer preparation, comprising:
- providing a plurality of glucose, mannose, and/or galactose containing glycan subunits (e.g., monomers or dimers);
- contacting the plurality of glycan subunits with a glycosidase enzyme selected from one of GT2, GT4, GT5, GT35, GT51, GH1, GH2, GH3, GH4, GH13.0, GH13.9, GH13.31, GH18, GH23, GH25, GH28, GH31, GH32, GH36, GH51, GH73, GH77, or GH94 CAZy family,
- under conditions that result in making a glycan polymer preparation, wherein a glycan polymer of the preparation is a substrate for a human gut microbe comprising a glycosidase enzyme of a GT2, GT4, GT5, GT35, GT51, GH1, GH2, GH3, GH4, GH13.0, GH13.9, GH13.31, GH18, GH23, GH25, GH28, GH31, GH32, GH36, GH51, GH73, GH77, GH94 CAZy family.
A method of making a glycan polymer preparation, comprising:
- providing a plurality of xylose, arabinose, galactose and/or glucose containing glycan subunits (e.g., monomers or dimers);
- contacting the plurality of glycan subunits with a glycosidase enzyme selected from one of GH13 subfamily 3, GH13 subfamily 30, GH30 subfamily 2, GH30 subfamily 5, GH43 subfamily 22, GH43 subfamily 8, or GH84 CAZy family,
- under conditions that result in making a glycan polymer preparation, wherein a glycan polymer of the preparation is a substrate for a human gut microbe comprising a glycosidase enzyme of a GH13 subfamily 3, GH13 subfamily 30, GH30 subfamily 2, GH30 subfamily 5, GH43 subfamily 22, GH43 subfamily 8, or GH84 CAZy family.
A method of making a glycan polymer preparation, comprising:
- providing a plurality of xylose, arabinose, galactose and/or glucose containing glycan subunits (e.g., monomers or dimers);
- contacting the plurality of glycan subunits with a glycosidase enzyme selected from one of GH3, GH106, GH105, GH2, GH20, GH28, GH76, GH97, or GH92 CAZy family,
- under conditions that result in making a glycan polymer preparation, wherein a glycan polymer of the preparation is a substrate for a human gut microbe comprising a glycosidase enzyme of a GH3, GH106, GH105, GH2, GH20, GH28, GH76, GH97, or GH92 CAZy family.
A method of making a glycan polymer preparation, comprising:
- providing a plurality of glucose and/or sialic acid containing glycan subunits (e.g., monomers or dimers);
- contacting the plurality of glycan subunits with a glycosidase enzyme selected from one of GH13 subfamily 19, GH13 subfamily 21, GH23, GH33, GH37 or GH104 CAZy family,
- under conditions that result in making a glycan polymer preparation, wherein a glycan polymer of the preparation is a substrate for a human gut microbe comprising a glycosidase enzyme of a GH13 subfamily 19, GH13 subfamily 21, GH23, GH33, GH37 or GH104 CAZy family.
A method of making a glycan polymer preparation, comprising:
- providing a plurality of glucose and/or sialic acid containing glycan subunits (e.g., monomers or dimers);
- contacting the plurality of glycan subunits with a glycosidase enzyme selected from one of GH23, GH24, or GH33 CAZy family,
- under conditions that result in making a glycan polymer preparation, wherein a glycan polymer of the preparation is a substrate for a human gut microbe comprising a glycosidase enzyme of a GH23, GH24, or GH33 CAZy family.
A method of making a glycan polymer preparation, comprising:
- providing a plurality of glucose, xylose, mannose, arabinose, and/or galactose containing glycan subunits (e.g., monomers or dimers);
- contacting the plurality of glycan subunits with a glycosidase enzyme selected from one of GH13 subfamily 20, GH13 subfamily 31, GH13 subfamily 39, GH39, GH43 subfamily 11, GH5 subfamily 44, or GH94 CAZy family,
- under conditions that result in making a glycan polymer preparation, wherein a glycan polymer of the preparation is a substrate for a human gut microbe comprising a glycosidase enzyme of a GH13 subfamily 20, GH13 subfamily 31, GH13 subfamily 39, GH39, GH43 subfamily 11, GH5 subfamily 44, or GH94 CAZy family.
A method of making a glycan polymer preparation, comprising:
- providing a plurality of glucose, xylose, mannose, arabinose, and/or galactose containing glycan subunits (e.g., monomers or dimers);
- contacting the plurality of glycan subunits with a glycosidase enzyme selected from one of GH2, GH31, GH23, GH13, or GH24 CAZy family,
- under conditions that result in making a glycan polymer preparation, wherein a glycan polymer of the preparation is a substrate for a human gut microbe comprising a glycosidase enzyme of a GH2, GH31, GH23, GH13, or GH24 CAZy family.
A method of making a glycan polymer preparation, comprising:
- providing a plurality of glucose, xylose, arabinose, and/or galactose containing glycan subunits (e.g., monomers or dimers);
- contacting the plurality of glycan subunits with a glycosidase enzyme selected from one of GH13 subfamily 3, GH13 subfamily 30, GH121, GH15, GH43 subfamily 27, GH43 subfamily 34, or GH43 subfamily 8 CAZy family,
- under conditions that result in making a glycan polymer preparation, wherein a glycan polymer of the preparation is a substrate for a human gut microbe comprising a glycosidase enzyme of a GH13 subfamily 3, GH13 subfamily 30, GH121, GH15, GH43 subfamily 27, GH43 subfamily 34, or GH43 subfamily 8 CAZy family.
A method of making a glycan polymer preparation, comprising:
- providing a plurality of glucose, xylose, arabinose, and/or galactose containing glycan subunits (e.g., monomers or dimers);
- contacting the plurality of glycan subunits with a glycosidase enzyme selected from one of GH92, GH97, GH76, GH28, GH20, GH105, GH2, GH50, GH3, or GH106 CAZy family,
- under conditions that result in making a glycan polymer preparation, wherein a glycan polymer of the preparation is a substrate for a human gut microbe comprising a glycosidase enzyme of a GH92, GH97, GH76, GH28, GH20, GH105, GH2, GH50, GH3, or GH106 CAZy family.
A method of making a glycan polymer preparation, comprising:
- providing a plurality of glucose, mannose, and/or galactose containing glycan subunits (e.g., monomers or dimers);
- contacting the plurality of glycan subunits with a glycosidase enzyme selected from one of GT11, GT10, GH92, GH51, GH35, GH29, GH28, GH20, GH130, GH13 subfamily 8, GH13 subfamily 14 CAZy family
- under conditions that result in making a glycan polymer preparation, wherein a glycan polymer of the preparation is a substrate for a human gut microbe comprising a glycosidase enzyme of a GT11, GT10, GH92, GH51, GH35, GH29, GH28, GH20, GH130, GH13 subfamily 8, GH13 subfamily 14 CAZy family.
A method of making a glycan polymer preparation, comprising:
- providing a plurality of glucose, mannose, and/or galactose containing glycan subunits (e.g., monomers or dimers);
- contacting the plurality of glycan subunits with a glycosidase enzyme selected from one of GT2, GT4, GH2, GH23, GH3, GT8, GT51, GT9, GH1, GH92, GH73, GH31, GH20, GH28, GT25, GT28, GT35, GH18, GT0, GH13, GH36, GH97, GH105, GH25, GH4, GH32, GH78, GH29, GH0, GH51, GT10, GH77 CAZy family,
- under conditions that result in making a glycan polymer preparation, wherein a glycan polymer of the preparation is a substrate for a human gut microbe comprising a glycosidase enzyme of a GT2, GT4, GH2, GH23, GH3, GT8, GT51, GT9, GH1, GH92, GH73, GH31, GH20, GH28, GT25, GT28, GT35, GH18, GT0, GH13, GH36, GH97, GH105, GH25, GH4, GH32, GH78, GH29, GH0, GH51, GT10, GH77 CAZy family.
A method of making a glycan polymer preparation, comprising:
- providing a plurality of xylose, arabinose, fucose and/or rhamnose containing glycan subunits (e.g., monomers or dimers);
- contacting the plurality of glycan subunits with a glycosidase enzyme selected from one of GT11, GT10, GH92, GH51, GH35, GH29, GH28, GH20, GH130, GH13 subfamily 8, GH13 subfamily 14 CAZy family
- under conditions that result in making a glycan polymer preparation, wherein a glycan polymer of the preparation is a substrate for a human gut microbe comprising a glycosidase enzyme of a GT11, GT10, GH92, GH51, GH35, GH29, GH28, GH20, GH130, GH13 subfamily 8, GH13 subfamily 14 CAZy family.
A method of making a glycan polymer preparation, comprising:
- providing a plurality of glycan subunits of a substrate of column E of Table 23, e.g., monomers or dimers;
- contacting the plurality of glycan subunits of a substrate with a glycosidase enzyme of column A of the same row as the substrate;
- under conditions that result in making a glycan polymer preparation, e.g., conditions of columns F, G, H, I, J, K, and/or L of the same row as the substrate and glycosidase enzyme.
A method of making a glycan polymer preparation, comprising:
- providing a plurality of xylose, arabinose, fucose and/or rhamnose containing glycan subunits (e.g., monomers or dimers);
- contacting the plurality of glycan subunits with a glycosidase enzyme selected from one of GT2, GT4, GH2, GH23, GH3, GT8, GT51, GT9, GH1, GH92, GH73, GH31, GH20, GH28, GT25, GT28, GT35, GH18, GT0, GH13, GH36, GH97, GH105, GH25, GH4, GH32, GH78, GH29, GH0, GH51, GT10, GH77 CAZy family,
- under conditions that result in making a glycan polymer preparation, wherein a glycan polymer of the preparation is a substrate for a human gut microbe comprising a glycosidase enzyme of a GT2, GT4, GH2, GH23, GH3, GT8, GT51, GT9, GH1, GH92, GH73, GH31, GH20, GH28, GT25, GT28, GT35, GH18, GT0, GH13, GH36, GH97, GH105, GH25, GH4, GH32, GH78, GH29, GH0, GH51, GT10, GH77 CAZy family.
A method of making a glycan polymer preparation, comprising:
- providing a plurality of glucose and/or galactose containing glycan subunits (e.g., monomers or dimers);
- contacting the plurality of glycan subunits with a glycosidase enzyme selected from one of GT3, GH97, GH43 subfamily 24, GH27, GH133, GH13 subfamily 8, GH13 CAZy family,
- under conditions that result in making a glycan polymer preparation, wherein a glycan polymer of the preparation is a substrate for a human gut microbe comprising a glycosidase enzyme of a GT3, GH97, GH43 subfamily 24, GH27, GH133, GH13 subfamily 8, GH13 CAZy family.
A method of making a glycan polymer preparation, comprising:
- providing a plurality of glucose and/or galactose containing glycan subunits (e.g., monomers or dimers);
- contacting the plurality of glycan subunits with a glycosidase enzyme selected from one of GT2, GT4, GH2, GH23, GH3, GT8, GT51, GT9, GH1, GH92, GH73, GH31, GH20, GH28, GT25, GT28, GT35, GH18, GT0, GH13, GH36, GH97, GH105, GH25, GH4, GH32, GH78, GH29, GH0, GH51, GT10, GH77, GT2, GT4, GH2, GH23, GH3, GT51, GH1, GT8, GH92, GT9, GH73, GH31, GH20, Gh28, GT35, GT28, GH18, GH13, GH97, GH25, GH36, GH4, GH105, GH32, GH78, GH29, GH0, GT25, GH51, GH77, GH88, GH24 CAZy family,
- under conditions that result in making a glycan polymer preparation, wherein a glycan polymer of the preparation is a substrate for a human gut microbe comprising a glycosidase enzyme of a GT2, GT4, GH2, GH23, GH3, GT8, GT51, GT9, GH1, GH92, GH73, GH31, GH20, GH28, GT25, GT28, GT35, GH18, GT0, GH13, GH36, GH97, GH105, GH25, GH4, GH32, GH78, GH29, GH0, GH51, GT10, GH77, GT2, GT4, GH2, GH23, GH3, GT51, GH1, GT8, GH92, GT9, GH73, GH31, GH20, Gh28, GT35, GT28, GH18, GH13, GH97, GH25, GH36, GH4, GH105, GH32, GH78, GH29, GH0, GT25, GH51, GH77, GH88, GH24 CAZy family.
A method of making a pharmaceutical composition that modulates a target human gut microbe, comprising
- providing a plurality of glycan subunits;
- contacting the glycan subunits of the plurality with a glycosidase enzyme composition having a glycosidase activity present in the target gut microbe, under conditions that result in the incorporation of the glycan subunits into a glycan polymer,
- optionally purifying the glycan polymer, and
- formulating the glycan polymer as a pharmaceutical composition for administration to the gut and modulation of the gut microbe, thereby making a pharmaceutical composition that modulates the target human gut microbe.
A purified preparation of glycosidase enzyme molecules comprising a glycosidase enzyme encoded by a nucleic acid sequence that is at least 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% identical to a nucleic acid sequence selected from one or more of SEQ ID NOs:
- 1-124,
- wherein the glycosidase enzyme is present in a human gut microbe.
A vector comprising a nucleic acid sequence that is at least 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% identical to a nucleic acid sequence selected from one or more of SEQ ID NOs:
- 1-124, wherein the nucleic acid encodes a glycosidase enzyme present in a human gut microbe, and wherein the vector is capable of being used to express the glycosidase enzyme.
A reaction mixture comprising:
- a glycosidase enzyme encoded by a nucleic acid sequence selected from one or more of SEQ ID NOs:
- 1-124, and a substrate, e.g., glycan subunits, e.g., monomers or dimers, of the glycosidase enzyme,
- wherein the substrate is present in a sufficient amount to form, e.g., by condensation, a glycan polymer.
Dependencies are inferred from the claim text.