Abstract: Provided are modified cellulase enzymes exhibiting increase cellulose-hydrolyzing activity in the presence of lignin and/or reduced binding to lignin comprising modified linker peptides comprising one or more amino acid substitutions, insertions, or deletions that result in (a) a decrease in the calculated isoelectric point of the linker peptide and/or (b) an increase in the ratio of threonine:serine in the linker peptide relative to a parental linker peptide from which said modified linker peptide is derived. Also provided are genetic constructs comprising nucleic acid sequences encoding for modified cellulase enzymes, methods for the production of the modified cellulase enzymes from host strains and a process for hydrolysing cellulose with the modified cellulases in the presence of lignin.

Abstract: Provided are modified cellulase enzymes exhibiting increase cellulose-hydrolyzing activity in the presence of lignin and/or reduced binding to lignin comprising modified linker peptides comprising one or more amino acid substitutions, insertions, or deletions that result in (a) a decrease in the calculated isoelectric point of the linker peptide and/or (b) an increase in the ratio of threonine:serine in the linker peptide relative to a parental linker peptide from which said modified linker peptide is derived. Also provided are genetic constructs comprising nucleic acid sequences encoding for modified cellulase enzymes, methods for the production of the modified cellulase enzymes from host strains and a process for hydrolysing cellulose with the modified cellulases in the presence of lignin.

Abstract: Provided are modified cellulase enzymes exhibiting increase cellulose-hydrolyzing activity in the presence of lignin and/or reduced binding to lignin comprising modified linker peptides comprising one or more amino acid substitutions, insertions, or deletions that result in (a) a decrease in the calculated isoelectric point of the linker peptide and/or (b) an increase in the ratio of threonine:serine in the linker peptide relative to a parental linker peptide from which said modified linker peptide is derived. Also provided are genetic constructs comprising nucleic acid sequences encoding for modified cellulase enzymes, methods for the production of the modified cellulase enzymes from host strains and a process for hydrolyzing cellulose with the modified cellulases in the presence of lignin.

Abstract: The present invention relates to a modified Family 5 cellulase comprising a substitution of an amino acid at position 363 with a non-native alanine, serine or threonine, the position determined from alignment of the modified Family 5 cellulase with amino acids 71-397 of a Trichoderma reesei Cel5A amino acid sequence as set forth in SEQ ID NO:1 and enzyme mixtures comprising same. Additionally provided is a genetic construct comprising a nucleic acid sequence encoding the modified Family 5 cellulase and a genetically modified microbe comprising the genetic construct. The invention also provides a process for producing the modified Family 5 cellulase.

Abstract: The present invention relates to a modified Family 5 cellulase comprising a substitution of an amino acid at position 363 with a non-native alanine, serine or threonine, said position determined from alignment of the modified Family 5 cellulase with amino acids 71-397 of a Trichoderma reesei Cel5A amino acid sequence as set forth in SEQ ID NO:1 and enzyme mixtures comprising same. Additionally provided is a genetic construct comprising a nucleic acid sequence encoding the modified Family 5 cellulase and a genetically modified microbe comprising the genetic construct. The invention also provides a process for producing the modified Family 5 cellulase.

Abstract: A modified Family 6 glycosidase enzyme comprising amino acid substitutions at one or more positions selected from the group 182, 367, 399, 400 and 427 is provided (the position determined form alignment of a parental Family 6 glycosidase with SEQ ID NO: 1). Genetic constructs and genetically modified microbes comprising nucleic sequences encoding the modified Family 6 glycosidase are also provided. Family 6 glycosidase of the invention display decreased hydrolysis activity of beta 1-4 linked polysaccharides and increased hydrolysis activity of beta 1-3, 1-4 linked polysaccharides compared with a parental Family 6 glycosidase. Such glycosidases find use in a variety of applications in industry, e.g., in hydrolysis of beta 1-3, 1-4 linked polysaccharides during the processing of cereal grains or the production of alcohol, animal feed or food products.

Abstract: A modified Family 6 cellulase enzyme comprising a proline residue at position 413 is provided. Genetic constructs and genetically modified microbes comprising DNA sequences encoding the modified Family 6 cellulase are also provided. Family 6 cellulases of the invention display improved thermostability, thermophilicity, alkalophilicity, or a combination thereof, relative to the parent Family 6 cellulases. Such cellulases find use in a variety of applications in industry that require cellulase stability and activities at temperatures, pH values, or both, above that of the native enzyme.

Abstract: A Family 6 cellulase variant enzyme comprising one or more than one amino acid substitution selected from a basic, polar or non-polar amino acid at position 103, a valine or isoleucine at position 136, a tyrosine at position 186, a glutamic acid or glutamine at position 365 and a glutamine at position 410 is provided (said position determined form alignment of the parental Family 6 with SEQ ID NO: 1). Genetic constructs and genetically modified microbes comprising DNA sequences encoding the Family 6 cellulase variant are also provided. Family 6 cellulases of the invention display reduced inhibition by glucose relative to the parent Family 6 cellulases. Such cellulases find use in a variety of applications in industry, e.g., in the hydrolysis of pretreated lignocellulosic feedstock, that require cellulose activity in the presence glucose concentrations that would otherwise inhibit the activity of the parental enzyme.

Abstract: A modified Family 6 cellulase enzyme comprising a proline residue at position 413 is provided. Genetic constructs and genetically modified microbes comprising DNA sequences encoding the modified Family 6 cellulase are also provided. Family 6 cellulases of the invention display improved thermostability, thermophilicity, alkalophilicity, or a combination thereof, relative to the parent Family 6 cellulases. Such cellulases find use in a variety of applications in industry that require cellulase stability and activities at temperatures, pH values, or both, above that of the native enzyme.

Abstract: Provided are modified cellulase enzymes exhibiting increase cellulose-hydrolyzing activity in the presence of lignin and/or reduced binding to lignin comprising modified linker peptides comprising one or more amino acid substitutions, insertions, or deletions that result in (a) a decrease in the calculated isoelectric point of the linker peptide and/or (b) an increase in the ratio of threonine:serine in the linker peptide relative to a parental linker peptide from which said modified linker peptide is derived. Also provided are genetic constructs comprising nucleic acidsequences encoding for modified cellulase enzymes, methods for the production of the modified cellulase enzymes from host strains and a process for hydrolysing cellulose with the modified cellulases in the presence of lignin.

Abstract: A modified Family 6 cellulase enzyme comprising a proline residue at position 413 is provided. Genetic constructs and genetically modified microbes comprising DNA sequences encoding the modified Family 6 cellulase are also provided. Family 6 cellulases of the invention display improved thermostability, thermophilicity, alkalophilicity, or a combination thereof, relative to the parent Family 6 cellulases. Such cellulases find use in a variety of applications in industry that require cellulase stability and activities at temperatures, pH values, or both, above that of the native enzyme.

Abstract: A modified Family 6 glycosidase enzyme comprising amino acid substitutions at one or more positions selected from the group 182, 367, 399, 400 and 427 is provided (the position determined form alignment of a parental Family 6 glycosidase with SEQ ID NO: 1). Genetic constructs and genetically modified microbes comprising nucleic sequences encoding the modified Family 6 glycosidase are also provided. Family 6 glycosidase of the invention display decreased hydrolysis activity of beta 1-4 linked polysaccharides and increased hydrolysis activity of beta 1-3, 1-4 linked polysaccharides compared with a parental Family 6 glycosidase. Such glycosidases find use in a variety of applications in industry, e.g., in hydrolysis of beta 1-3, 1-4 linked polysaccharides during the processing of cereal grains or the production of alcohol, animal feed or food products.

Abstract: A Family 6 cellulase variant enzyme comprising one or more than one amino acid substitution selected from a basic, polar or non-polar amino acid at position 103, a valine or isoleucine at position 136, a tyrosine at position 186, a glutamic acid or glutamine at position 365 and a glutamine at position 410 is provided (said position determined form alignment of the parental Family 6 with SEQ ID NO: 1). Genetic constructs and genetically modified microbes comprising DNA sequences encoding the Family 6 cellulase variant are also provided. Family 6 cellulases of the invention display reduced inhibition by glucose relative to the parent Family 6 cellulases. Such cellulases find use in a variety of applications in industry, e.g., in the hydrolysis of pretreated lignocellulosic feedstock, that require cellulose activity in the presence glucose concentrations that would otherwise inhibit the activity of the parental enzyme.

Abstract: A modified Family 6 cellulase enzyme comprising a proline residue at position 413 is provided. Genetic constructs and genetically modified microbes comprising DNA sequences encoding the modified Family 6 cellulase are also provided. Family 6 cellulases of the invention display improved thermostability, thermophilicity, alkalophilicity, or a combination thereof, relative to the parent Family 6 cellulases. Such cellulases find use in a variety of applications in industry that require cellulase stability and activities at temperatures, pH values, or both, above that of the native enzyme.

Abstract: Endosteal fixation of a ligament graft with an interference fixation device installed in a retrograde manner in ACL reconstruction is disclosed. The interference fixation device may be a screw or a ribbed implant fitted with a length of reinforced suture or wire for pulling the interference fixation device through the tibial tunnel. The suture extends beyond the leading tip of the interference fixation device a sufficient length to allow the interference fixation device to be passed into the joint by pulling on the suture or wire exiting the tibial tunnel. A driver or an impaction device, such as a slap hammer, is employed to advance and insert the interference fixation device in a retrograde manner into the tibial tunnel. Accordingly, interference fixation of the graft near the tibial plateau is provided, thereby eliminating graft abrasion at the tibial plateau tunnel opening.

Abstract: Endosteal fixation of a ligament graft with an interference fixation device installed in a retrograde manner in ACL reconstruction is disclosed. The interference fixation device may be a screw or a ribbed implant fitted with a length of reinforced suture or wire for pulling the interference fixation device through the tibial tunnel. The suture extends beyond the leading tip of the interference fixation device a sufficient length to allow the interference fixation device to be passed into the joint by pulling on the suture or wire exiting the tibial tunnel. A driver or an impaction device, such as a slap hammer, is employed to advance and insert the interference fixation device in a retrograde manner into the tibial tunnel. Accordingly, interference fixation of the graft near the tibial plateau is provided, thereby eliminating graft abrasion at the tibial plateau tunnel opening.