Glycosyl hydrolase enzymes and uses thereof
The present disclosure is generally directed to glycosyl hydrolase enzymes, compositions comprising such enzymes, and methods of using the enzymes and compositions, for example for the saccharification of cellulosic and hemicellulosic materials into sugars.
1. A non-naturally occurring composition comprising:
(a) a first polypeptide having f3-xylosidase activity, wherein the first polypeptide has at least 90% sequence identity to the amino acid sequence of SEQ ID NO:28 or the amino acid sequence corresponding to residues (i) 20-341, (ii) 21-350, (iii) 107-341, or (iv) 107-350 of SEQ ID NO:28; and
(b) a second polypeptide having xylanase activity;
wherein the non-naturally occurring composition comprises one or more polypeptides produced from a host cell, wherein the host cell is engineered to express the first polypeptide having β-xylosidase activity and wherein the first polypeptide is heterologous to the host cell.
2. The composition of claim 1 , wherein the second polypeptide is a Trichoderma xylanase or an Aspergillus xylanase.
3. The composition of claim 2 , wherein the Trichoderma xylanase is a Trichoderma reesei Xyn3 polypeptide or a Trichoderma reesei Xyn2 polypeptide, or the Aspergillus xylanase is an Aspergillus fumigatus AfuXyn2 polypeptide or an Aspergillus fumigatus AfuXyn5 polypeptide, wherein:
said Trichoderma reesei Xyn3 polypeptide has at least 90% sequence identity to SEQ ID NO:42, or to residues 17-347 of SEQ ID NO:42, or is encoded by a nucleic acid having at least 90% sequence identity to SEQ ID NO:41;
said Trichoderma reesei Xyn2 polypeptide has at least 90% sequence identity to SEQ ID NO:43, or to residues 33-222 of SEQ ID NO:43;
said Aspergillus fumigatus AfuXyn2 polypeptide has at least 90% sequence identity to SEQ ID NO:24, or to residues 19-228 of SEQ ID NO:24, or is encoded by a nucleic acid having at least 90% sequence identity to SEQ ID NO:23;
said Aspergillus fumigatus AfuXyn5 polypeptide has at least 90% sequence identity to SEQ ID NO:26, or to residues 20-313 of SEQ ID NO:26, or is encoded by a nucleic acid having at least 90% sequence identity to SEQ ID NO:25.
4. The composition of claim 1 , further comprising one or more accessory proteins selected from the group consisting of mannanases, galactanases, arabinases, ligninases, amylases, glucuronidases, proteases, esterases, lipases, xyloglucanases, glycoside hydrolase Family 61 polypeptides, CIP1, CIP2, swollenin, expansins, CIP1-like proteins, CIP2-like proteins, cellobiose dehydrogenases, and manganese peroxidases.
5. The composition of claim 1 , further comprising a biomass.
6. The composition of claim 5 , wherein the biomass is pretreated biomass, optionally wherein the biomass is ammonia-pretreated.
7. The composition of claim 6 , wherein the combined weight of polypeptides having xylanase activity is 0.5 g to 40 g per 1 kg of hemicellulose in the biomass, or the combined weight of polypeptides having f3-xylosidase activity is 0.5 g to 50 g per 1 kg of hemicellulose in the biomass.
8. The composition of claim 7 , wherein the biomass comprises one or more of corncob, corn stover, corn fiber, switchgrass, sorghum, paper, pulp, sugarcane bagasse, or Miscanthus.
9. A fermentation broth comprising the composition of claim 1 .
10. The fermentation broth of claim 9 , which is the fermentation broth of a filamentous fungus, wherein the filamentous fungus is a Trichoderma, Humicola, Fusarium, Aspergillus, Myceliophthora, Neurospora, Penicillium, Cephalosporium, Achlya, Podospora, Endothia, Mucor, Cochliobolus, Pyricularia, Phanerochaete , or Chrysosporium.
11. A saccharification process comprising treating a material comprising hemicellulose with the composition of claim 1 .
12. The process of claim 11 , wherein the material comprising hemicellulose is one or more of corncob, corn stover, corn fiber, switchgrass, sorghum, paper, pulp, sugarcane bagasse, or Miscanthus;
optionally wherein the process yields at least 60% xylose from hemicellulose xylan of the material comprising hemicellulose.