98 results about "Enzyme variant" patented technology

The present invention relates to lipases with a common three-dimensional structure, and variants thereof with an improved half-life improvement factor (HIF). The present invention also relates to polynucleotides encoding the lipases and variants, nucleic acid constructs, vectors, and host cells comprising the polynucleotides, compositions comprising the lipases or variants of the invention, and use of the variants.

The present invention is related to the field of protein engineering technology which increases the enzymatic activity and thermal stability of human hyaluronidase which is an enzyme that hydrolyzes hyaluronic acid; and more particularly to hyaluronidase PH20 variants or fragments thereof, which comprise one or more amino acid residue substitutions in the region corresponding to the alpha-helix region and / or its linker region in the amino acid sequence of wild-type PH20 of SEQ ID NO: 1 and in which one or more amino acid residues at the N-terminus and / or the C-terminus are selectively truncated additionally.Specifically, the present invention relates to PH20 variants or fragments thereof, which comprise one or more amino acid residue substitutions selected from the group consisting of T341A, T341C, T341G, S343E, M345T, K349E, L353A, L354I, N356E and I361T in wild-type PH20 having the amino acid sequence of SEQ ID NO: 1, and additionally comprise the substitution of amino acids located in the alpha-helix 8 region and / or a linker region between alpha-helix 7 and alpha-helix 8 in the amino acid sequence of wild-type PH20, and in which portion(s) of amino acids located at the N-terminal and C-terminal regions are deleted.

Compositions and methods are provided for variant Cas systems and elements comprising such systems, including, but not limiting to, Cas endonuclease variants, guide polynucleotide / Cas endonuclease complexes comprising Cas endonuclease variants, as well as guide polynucleotides and guide RNA elements that can interact with Cas endonuclease variants. Compositions and methods are provided for genome modification of a target sequence in the genome of a cell. The methods and compositions employ a guide polynucleotide / Cas endonuclease system comprising a Cas9 endonuclease variant to provide an effective system for modifying or altering target sequences within the genome of a cell or organism.

The present invention relates to a method for obtaining a detergent composition comprising introducing (a) a variant of a parent lipase having at least 60% sequence identity with SEQ ID NO:2, having a substitution at a position D254 of the mature polypeptide corresponding to SEQ ID NO:2, and having lipase activity; and (b) an anionic surfactant, wherein the composition has increased stability compared to a corresponding composition containing the parent lipase.

Provided is a lipase mutant with alleviated inhibition of activity against a dispersed substrate in the presence of a surfactant. The lipase mutant consists of an amino acid sequence having an identity of at least 75% with the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8 and has a predetermined amino acid residue at a predetermined position numbered according to SEQ ID NO: 2.

The present disclosure relates, according to some embodiments, to systems, apparatus, compositions, methods, and workflows that include salt active nucleases (e.g., salt active nuclease variants) with desirable properties including, for example, salt tolerance.

The present application relates to the field of biotechnology and enzyme engineering, and particularly relates to a lipase mutant expressed in Escherichia coli and application thereof. Specifically, in order to solve the technical problem that the catalytic activity of the existing Rhizopus oryzae lipase (ROL) is insufficient and it is difficult to meet the needs of industrial production, the purpose of the present application is to provide a lipase variant with significantly improved catalytic activity and / or thermal stability. On the basis of the existing ROL four-point mutant ROL-E190C / V209L / E238C / D262G (the amino acid sequence is shown as SEQ ID NO. 1), the present application innovatively introduces a machine learning assisted design strategy, and through multiple rounds of 'prediction-verification-model optimization' iteration cycles, a series of excellent lipase mutants are successfully obtained.

The invention provides a VniNatt enzyme mutant, nucleic acid, a carrier or a host cell, and application of the VniNatt enzyme mutant, nucleic acid, a carrier or a host cell in fermentation production of NMN. Compared with a wild type, the mutant can catalytically produce NMN with higher efficiency, and has good thermal stability. BL21 engineering bacteria are constructed on the basis of the VniNamt enzyme mutant and are used for producing NMN through fermentation, and the yield of the NMN can reach 54 g / L by adopting a fed-batch fermentation process.

According to the invention, beta-glycosidase which has a catalytic effect on vincoside, loganin and strictosamide is selected, and a foundation is laid for diversity and possibility of an aglycone production process. By researching the three-dimensional structure information of the Gelsemium elegans Beta-glycosidase, a zymoprotein structure information basis is provided for molecular modification of the Gelsemium elegans Beta-glycosidase, enzyme variants capable of improving substrate selectivity, thermal stability and catalytic efficiency are obtained more efficiently, and the requirements of industrial application are met; the invention provides a truncated mutant enzyme with improved enzymatic activity, and through induced expression and activity screening, it is found that the enzyme activity of the obtained mutant enzyme to loganin and strictosamide is about two times higher than that of a wild enzyme.

Provided herein are phenylalanine-degrading enzyme variants for use in degrading phenylalanine and in treating phenylketonuria (PKU). More specifically, provided herein are phenylalanine-degrading enzyme variants that exhibit increased thermal stability and physicochemical resistance as measured by residual phenylalanine-degrading activity following challenge.

A polypeptide comprising: a) milk-clotting enzyme activity, wherein the C / P ratio of clotting activity (C) to proteolytic activity (P) is equal to or greater than the C / P ratio of a polypeptide having the amino acid sequence of SEQ ID NO: 2; and b) alpha-S1-I casein fragment generating activity that is equal to or greater than the alpha-S1-I casein fragment generating activity of a polypeptide having the amino acid sequence of SEQ ID NO: 2; and / or c) a clotting time at pH 6.1 that is equal to or less than the clotting time of a polypeptide having the amino acid sequence of SEQ ID NO: 2.

The present invention relates to variants of a parent lipase, wherein said variants have at least 60% but less than 100% sequence identity to SEQ ID NO: 2, have lipase activity, the variants comprise a substitution at a position corresponding to position 92 and / or 96 of SEQ ID NO: 2; and are unchanged at positions corresponding to positions 231, 233 and 254 of SEQ ID NO: 2. The present invention also relates to polynucleotides encoding these variants, nucleic acid constructs, vectors, and host cells comprising the polynucleotides, and methods of using the variants.

A polypeptide having an amino acid sequence that, when compared with the amino acid sequence shown in SEQ ID NO:1, contains at least substitutions of amino acid residues corresponding to amino acids at sites 50, 51, 126, 135, and 221, wherein the sites are defined with reference to SEQ ID NO:1.

A sortase variant with improved catalytic activity and stability, in particular said mutant further comprises the mutations M155V / L200F or M155V / K162P, according to SEQ ID NO: 1, with respect to the parent sortase (SEQ ID NO: 2). Also the use of said sortase mutant is involved.

The current invention relates to a computer-implemented method for ranking and / or identifying substrate-enzyme combinations according to the likelihood that a substrate undergoes catalytic conversion by an enzyme or inhibits the enzyme; to a computer system; and a computer program product. The invention also relates to uses of the method for predicting the substrate range of an enzyme X, for screening enzyme variants with altered substrate specificity and / or catalytic activity in relation to a substrate of interest, and for predicting biological phenotypes associated with an enzyme X.

The present invention relates to a novel 5-dehydro-2-deoxygluconate kinase variant and a method for producing 5 '-inosinic acid using the same. According to the present invention, the 5-dehydro-2-deoxygluconate kinase variant has a modified protein activity due to the substitution of one or more amino acids in the amino acid sequence constituting a 5-dehydro-2-deoxygluconate kinase, and thus a recombinant microorganism comprising the 5-dehydro-2-deoxygluconate kinase variant can efficiently produce 5 '-inosinic acid.

Provided is a novel cis-aconitate synthesis enzyme, more particularly, a recombinant microorganism for producing itaconate including a cis-aconitate synthesis enzyme variant. According to the present disclosure, it was confirmed that the production and yield of itaconate were significantly increased in the recombinant microorganism for producing itaconate into which the novel cis-aconitate synthesis enzyme was introduced. In addition, it was confirmed that in the recombinant microorganism for producing itaconate of the present disclosure, a new carbon flow to itaconate was separated from the existing TCA cycle based on the activity of the corresponding enzyme. Accordingly, the novel aconitate synthesis enzyme of the present disclosure and the recombinant microorganism introduced with the aconitate synthesis enzyme can increase the economic feasibility of itaconate, and thus can be used in various industrial fields, such as synthetic resins, latexes, and food additives in which itaconate is used.

The present invention relates to new peptiligases, which are characterized by sequences comprising two amino acid substitutions, one at position 2 and one at position 206 compared to the peptiligases in the prior art, wherein the numbering is according to the numbering of the subtilisin BPN' enzyme, as well as to methods of producing these new peptiligase variants. It further relates to a method of improving the producibility of a peptiligase in a bacillus subtilis, without impacting negatively on its activity in a coupling reaction of peptide fragments.

The present invention relates to variants of a parent lipase which has lipase activity and comprise one or more substitutions corresponding to G23S, D27N, A40I, F51I,L, E56R, D57N, V60E,K, K98I, N101D, R118F, G163S, T231R, N233R, Y220F, T244E, and P256T using SEQ ID NO: 2 for numbering. The present invention also relates to compositions or microcapsule compositions comprising a lipase variant of the invention and to liquid products comprising a microcapsule composition of the invention as well as the use of said microcapsule composition for stabilizing lipase variants of the invention.

This invention provides libraries of Botulinum neurotoxin A (BoNT / A) protease variants for stepwise evolution of the enzyme to generate BoNT / A variants that can specifically cleave a desired cleavage site in a target protein. Related methods for performing stepwise evolution of BoNT / A with the BoNT / A variant libraries and simultaneous stepwise evolution of a desired substrate sequence in a target protein are also provided by the disclosure. Additionally provided in the disclosure are specifically evolved BoNT / A variant enzymes and conjugate proteins that specifically degrade intrinsically disordered proteins (IDPs) that are involved in human deceases, e.g., a-Synuclein. Polynucleotide sequences encoding the engineered BoNT / A proteases, expression vectors and related pharmaceutical compositions are also provided in this disclosure. Further encompassed by the invention are therapeutic methods that utilize the engineered BoNT / A enzymes in the treatment of various synucleinopathies.

The present invention relates to lipase variants with reduced activity at pHs around neutral compared to the parent. The present invention also relates to compositions comprising a lipase variant of the invention; polynucleotides encoding lipase variants of the invention; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of producing and using the variants for cleaning.

The present invention provides to an improved high-throughput system and method for generated and screening of genetic variants by combinatorial modifications. Also provided are optimized SpCas9 enzyme variants produced by this system.

The present invention relates to a method for estimating or determining fitness of enzyme variants and / or optimizing a nucleotide sequence encoding an enzyme variant comprising generating a DNA variant library and linking a unique DNA barcode to each DNA variant, wherein the DNA variants are subjected to long-read and short-read sequencing. In particular, the present invention relates to a method for predicting or determining fitness of enzyme variants and / or optimizing a nucleotide sequence encoding an enzyme variant by combining DNA barcodes, selection pressure, long-read sequencing, and short-read sequencing. Furthermore, the present invention relates to application of machine learning for predicting enzyme fitness and / or optimizing a nucleotide sequence encoding an enzyme variant based on data obtained from said method.

The present disclosure relates to an isopropylmalate synthase variant and a method of producing L-leucine using the same.

The present invention relates to a novel ABC transporter permease variant and a method for producing 5 '-inosinic acid using the same. The ABC transporter permease variant has a modified protein activity due to the substitution of one or more amino acids in the amino acid sequence constituting the ABC transporter permease, and thus a recombinant microorganism comprising the ABC transporter permease variant can efficiently produce 5 '-inosinic acid.

The present disclosure provides endonuclease variants having improved properties, such as hyperactivity and / or low indiscriminate single strand DNase activity, relative to the corresponding wild-type endonucleases.

The present invention provides a GG-specific mismatch endonuclease variant, a TT-specific mismatch endonuclease variant, and a GT / TG-specific mismatch endonuclease variant. The present invention also provides a mismatch specific cleaving reaction using said variant, a method for removing errors in a nucleic acid amplification reaction using a mismatch nuclease, a method for suppressing amplification of a nucleic acid having a specific base sequence during a nucleic acid amplification reaction, and a method for detecting a nucleic acid having a single base polymorphic mutation using said suppression method.

The present disclosure relates to modified RNA ligase polypeptides and compositions thereof, as well as polynucleotides encoding the modified RNA ligase polypeptides. The disclosure also provides methods of using the modified RNA ligase polypeptides or compositions thereof for molecular biology, diagnostic, and other purposes. In some embodiments, the amino acid sequence of the modified RNA ligase comprises at least one substitution as provided in Tables 5.1, 6.1, 7.1, 8.1, 9.1, 10.1, 11.1, and 12.1, where the amino acid positions are relative to a reference sequence corresponding to residues 12-346 of SEQ ID NO:14 or a reference sequence corresponding to SEQ ID NO:14.

The present invention relates to a protein engineering technology that enhances the enzyme activity of human hyaluronidase, a hyaluronic acid hydrolase, at neutral pH, and more particularly to a novel hyaluronidase mutant or section thereof, comprising one or more amino acid residue substitutions in or near the enzyme active site of the amino acid sequence of the natural hyaluronidase Hyal1 (SEQ ID NO: 1), and selectively cleaving additional N-terminal and / or C-terminal amino acid residues.

The present invention relates to a synthetic phytase which comprises (a) the amino acid sequence as set forth in SEQ ID NO. 1, (b) a variant of (a) having at least 70% sequence identity to SEQ ID NO. 1, or (c) a fragment, fraction or shuffled variant of (a) or (b) maintaining phytase activity, with the proviso that the phytase comprises one or more amino acid substitutions at one or more positions relative to SEQ ID NO. 1 selected from the group consisting of V13, T30, N37, K45, L46, T81, N83, A89, A113, Q121, Q122, A123, H128, K131, K139, A142, H143, Q162, S164, A166, S170, Q182, N184, G186, K187, L188, A194, M195, A200, I201, N202, D204, N206, K207, A209, S218, T219, L225, H228, K234, N239, E243, S248, Q256, F257, M260, S261, N270, P288, P292, A311, A314, L319, S320, W321, K344, K347, M355, L359, Q365, P367, E372, T388, Q404, and N405.