163 results about "Enzyme variant" patented technology

The invention includes a mutant T7 DNA ligase or a biologically active fragment thereof, which has greater activity than wild type T7 DNA ligase. The mutant T7 DNA ligase, or the biologically active fragment, has one or more substitutions differing from the wild type, as described more fully in the Summary. The preferred mutant T7 DNA ligase has at least one of the following mutations: E63K (SEQ ID NO:4), K73E (SEQ ID NO:6), K137E (SEQ ID NO:7), K174E (SEQ ID NO:9), E182K (SEQ ID NO:11), K210E (SEQ ID NO: 13), E243K (SEQ ID NO:15), D245R (SEQ ID NO: 17), E268K (SEQ ID NO:19), E272K (SEQ ID NO:21), E289K (SEQ ID NO:23), K295E (SEQ ID NO:25) and D336R (SEQ ID NO:27).

There is provided a fluorinase variant thereof having at least 70% sequence identity to a sequence MSDLGX6TDDSVAQCKGLMLSICPX24VX26IX28DX30CHX33MTPX37DVVEGARYIVDLPR X52FPEGTVFATTTYPATGTX70X71RSVAX76RX78KX80AALGGARGQX90AGSGX95GX97E RAEGX103YIYIAPNNGLLTX116VIEEHGYX124EAYEVSX131TX133VIPX137X138PEPTFYSR EMVAIPSAHLAAGFPLX163X164VGRX168LX170DX172EIVRFEX179X180KX182X183X184VX186G X188X189LX191GX193X194X195X196X197DHPFGNX204WTNX208HRTDLEKAGIX219YX221TX223X 224KX226VX228DGVLX233FX235LPLX239PTFADAX246X247X248GX250PVX253YX255NSRGYLX2 62X263ARNAAX269LAYPYNLX277AGX280SVX283VTX286A (SEQ ID NO: 1), wherein X is a natural amino acid, and wherein the variant comprises one or more mutations at a site selected from the group consisting of an ion-egress site, a S-Adenosyl-L-Methionine (SAM) binding site, an ion-binding site (IBS), and a conserved site. Also disclosed are polynucleotides encoding the variants, vectors comprising the polynucleotides encoding the variants, and host cells comprising the vectors thereof. Also disclosed are Methods for producing the variants, methods of catalyzing the fluorination of a compound, uses of the fluorinase variants, and methods of treatment using the fluorinase variants thereof.

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 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 cleaved 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 one or more amino acids located at the N-terminal and C-terminal regions are deleted.

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 relates to variants of Terminal deoxynucleotidyl Transferase (TdT), each of which (i) has an amino acid sequence similarity to SEQ ID NO: 2, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33 or 35 with corresponding amino acid substitutions, (ii) is capable of synthesizing a nucleic acid fragment without a template and (iii) is capable of incorporating a modified nucleotide into the nucleic acid fragment.

The present invention is directed to terminal deoxynucleotidyltransferase (TdT) variants that (i) comprise an amino acid sequence that is at least a specified percent identical to an indicated SEQ ID NOs and have at least one substitution at Q455 or at least Q455 plus at least one further substitution at G186, S248, T331, Q390, K394 or H466 (where positions are with respect to SEQ ID NO 1 and functionally equivalent positions in indicated SEQ ID NOs), (ii) are capable of template-free extension of a polynucleotide, and (iii) exhibit enhanced stability or enhanced efficiency in incorporating 3′-0-blocked nucleoside triphosphates into a polynucleotide. The invention is also directed to the use of these TdT variants for synthesizing polynucleotides of any predetermined sequence.

The invention discloses a trichoderma reesei engineering bacterium for secreting and expressing egg white lysozyme and a construction method of the trichoderma reesei engineering bacterium, and belongs to the field of molecular biology and biotechnology. The invention firstly provides an egg white lysozyme mutant, and the amino acid sequence of the egg white lysozyme mutant is shown as any one of SEQ ID NO.2-4. The construction method of the trichoderma reesei engineering bacterium for secreting and expressing the egg white lysozyme comprises the following steps: by taking trichoderma reesei as a starting strain, transforming an expression module into the trichoderma reesei to construct the trichoderma reesei engineering bacterium; the expression module comprises a promoter, a secretory peptide gene, an egg white lysozyme gene and a terminator which are connected in sequence. The coding gene of the egg white lysozyme is transformed into a trichoderma reesei genome, so that a trichoderma reesei engineering strain for secreting and expressing the egg white lysozyme is successfully constructed. Meanwhile, different egg white lysozyme variants can bring different effects, and when the trichoderma reesei RUT-C30 is used as an original strain, the effect is superior to that of QM6a.

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.

Lipase variants of rhizomucor miehei lipase are described herein. The lipase has a peptide sequence having at least 80% sequence similarity to SEQ ID NO: 2 and having an F94 mutation of SEQ ID NO: 2. Also described is a method of increasing the expression level of a rhizomucor miehei lipase having SEQ ID NO: 2 by introducing an F94 mutation of SEQ ID NO: 2 to produce a rhizomucor miehei lipase variant comprising a peptide sequence having at least 80% similarity to SEQ ID NO: 2 and having an F94 mutation of SEQ ID NO: 2; and the rhizomucor miehei lipase variant is expressed in a protein production system. The lipases described herein are useful for hydrolyzing an ester by providing a mixture containing a lipase variant and an ester; and a reaction product is formed under proper reaction conditions.

The present invention relates to novel glycoside synthetases for antibody drug complex (ADC) engineering. Enzyme variants, referred to as EndoSd-D232M and EndoSz-D234M, contain a polysaccharide binding and / or modification activity at the conserved N297 glycosylation site of the Fc region of an exemplary antibody. It has been proved that the glycoside synthase activity of EndoSd-D232M and EndoSz-D234M can be applied to various mAbs with different receptors as targets, including but not limited to Globo H, SSEA-4 and SSEA-3 series receptors (OBI-888, Globo H ganglioside), Herceptin (Her2 receptor), Perjeta (Her2 receptor) and Vectibix (EGFR receptor). It has been found that both the mAb-GlcNAc and the mAb-GlucNAc (F) are suitable receptors for both EndoSd-D232M and EndoSz-D234M. The invention also relates to a method for preparing the same.

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 generally to polypeptides capable of hydrolyzing amido linkages in polyamides, and more particularly to polypeptides comprising the amino acid sequence of amino acid residues 2-398 of SEQ ID NO: 2 or an amino acid sequence having at least 70% sequence identity thereto; or a polypeptide comprising the amino acid sequence of amino acid residues 2-394 of SEQ ID NO: 88 or an amino acid sequence having at least 75% sequence identity thereto. The disclosure also extends to methods of using the polypeptides. In another embodiment, the disclosed polypeptides are capable of hydrolyzing nylon 6, 6 oligomers to produce adipic acid and hexamethylenediamine.

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.

A number of T4 DNA ligase mutants exhibiting enhanced ligation activity in the presence of high salt concentrations compared to the wild-type ligase were engineered, characterized, and selected via gel electrophoresis of ligation products from a standard ligation assay. Ligase catalyzes the formation of phosphodiester bonds between the 5′ and 3′ ends of complementary cohesive ends or blunt ends of duplex DNA, a process that is vital to numerous molecular biology processes including cloning and sequencing.

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.

The invention provides a telomerase variant and application of the telomerase variant in anti-aging. Furthermore, compared with the original telomerase, the activity of the telomerase variant is further enhanced, and the improved telomerase variant can more remarkably improve the anti-aging effect. The composition provided by the invention is substantially an anti-aging composition, the composition comprises a high-activity telomerase mutant and a specific auxiliary agent, and the composition has relatively high biological activity, relatively good application prospects and is suitable for marketization.

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.

The invention belongs to the technical field of gene editing, and discloses a COME nuclease variant and application thereof. The invention provides a method for controlling oligomerization engineering. An oligomerization structural domain is integrated in CRISPR (clustered regularly interspaced short palindromic repeats) nuclease. A new optimization path is created by controlling the three-dimensional space arrangement of the CRISPR protein compound instead of only modifying the internal catalytic characteristics of the CRISPR protein compound; the traditional balance between the efficiency and the specificity can be broken through, and the efficiency and the specificity are remarkably improved at the same time; the method can be universally applied to a general enhancement platform of various CRISPR systems with different evolutionary aspects; synergistic interaction with the existing optimization method is realized, and a comprehensive optimization effect exceeding the sum of effects of a single method is generated.

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 application discloses a multiplex method for measuring nucleic acid modification enzyme activity and screening one or more variable elements of enzymatic reaction.The method includes: (i) providing a polynucleotide construct library encoding DNA / RNA target site and variable element to be tested such as nucleic acid modification enzyme variant, (ii) partitioning a single copy of the polynucleotide construct with in vitro transcription and translation (IVVT) reagent, (iii) allowing the IVVT reaction of nucleic acid modification enzyme and DNA / RNA target site to be expressed in each compartment, (iv) depending on the functionality of the nucleic acid modification enzyme encoded, single polynucleotide construct / RNA target is cracked, remains intact or otherwise modified, and (v) by sequencing to cracked, complete or modified polynucleotide construct / RNA target quantification, so as to directly identify the enzymatic activity related to each variable element and to the enzymatic activity quantification.Particularly, nucleic acid modification enzyme is CRISPR associated protein (Cas).

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.