402 results about "Protein engineering" patented technology

The invention provides a recombinant XVII type collagen and application thereof, and relates to the technical field of protein engineering. The novel recombinant human XVII type collagen is obtained by optimizing and screening collagen structural domain sequences of the human XVII type collagen and then combining the sequences. The recombinant human XVII type collagen is consistent with the corresponding part of the amino acid sequence of the human XVII type collagen, does not generate immunological rejection when being applied to a human body, and can be widely applied to the industries of surgical medical treatment and medical beauty. In addition, the recombinant human XVII type collagen is high in stability, and the problem of protein degradation in the process of expressing recombinant protein by pichia pastoris can be avoided. In addition, the recombinant human XVII type collagen also has the effects of high biological activity, high biological safety, promotion of cell proliferation and differentiation and promotion of hair growth, and can be applied to the field of cosmetics or medicines as a functional component.

The invention provides a micromolecular recombinant XVII type collagen as well as a preparation method and application thereof, and relates to the technical field of protein engineering. The novel recombinant human XVII type collagen is obtained by optimizing and screening a plurality of spiral region sequences such as a collagen structural domain, a C-terminal region and a middle region of the human XVII type collagen. The amino acid composition of the recombinant XVII type collagen is consistent with that of a natural collagen alpha1 chain, immunological rejection is not generated when the recombinant XVII type collagen is applied to a human body, the recombinant XVII type collagen can be widely applied to surgical medical treatment and medical beauty industries, the molecular weight of the collagen is small, efficient secretion soluble expression in eukaryotic host cells such as pichia pastoris can be achieved, and the recombinant XVII type collagen has a good application prospect. And industrial large-scale production is carried out. According to the present invention, the verification results show that the micromolecular recombinant XVII type collagen further has basement membrane anti-aging activity, wherein the basement membrane anti-aging activity specifically comprises the improvement of the expression levels of the IV type collagen, the VII type collagen and the XVII type collagen;

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 belongs to the technical field of protein engineering. Provided are a recombinant type XVII collagen having a triple helix structure and the use thereof. The amino acid sequence of the recombinant type XVII collagen comprises n core units, wherein n is the integer 1 or an integer greater than 1, and the amino acid sequence of the core units is as shown in SEQ ID NO: 1. Further provided are a method for constructing a plurality of expression systems and a method for preparing the recombinant type XVII collagen. Further provided is the actual industrial application on the basis of the characteristics of the protein. The provided recombinant type XVII collagen exhibits high purity, good thermal stability and high biological activity, and has a molecular weight much smaller than that of natural collagen, thereby reducing the difficulty for the collagen to cross the skin barrier and exert biological functions. Experiments show that the recombinant type XVII collagen has a variety of biological activities such as promoting cell migration.

The invention discloses a recombinant human IV-type collagen as well as a preparation method and application thereof, and belongs to the field of bioengineering and protein engineering. According to the invention, three recombinant human IV type collagen new molecules are designed and screened on the basis of a part of sequences in a human IV type collagen alpha1 chain, soluble expression of a target protein is realized by using an escherichia coli system, and the recombinant collagen with the purity of more than 98% can be obtained through nickel column purification and acid regulation treatment. The recombinant collagen provided by the invention has good cell migration promotion activity and cell adhesion activity, and can be widely applied to the fields of tissue engineering, medical equipment, cosmetics, health care products and the like.

The present disclosure provides a recombinant type XVII collagen with a triple helical structure and use thereof and belongs to the field of protein engineering technology. The recombinant type XVII collagen has an amino acid sequence formed by tandem repeats of n core units, wherein n is an integer of 1 or greater than 1, and the core unit has the amino acid sequence as set forth in SEQ ID NO: 1. The present disclosure also provides methods for constructing multiple expression systems and methods for preparing the recombinant type XVII collagen.

The invention provides recombinant IV type collagen and application thereof, and relates to the technical field of protein engineering. In the invention, an amino acid residue repetitive unit with good biological activity and stability in the human-derived IV-type collagen is found, and the amino acid residue repetitive unit is used as a core for repeating and constructing the recombinant IV-type collagen, so that the novel human-derived recombinant IV-type collagen with biological activity is obtained. The human-derived recombinant IV-type collagen provided by the invention is a small-molecular-weight protein, is easy to absorb by a human body, is good in safety, can remarkably promote secretion of collagen, also has the effects of promoting cell adhesion and cell migration, and has a good application prospect in the aspects of basement membrane repair and remodeling. A new functional component is provided for products for repairing and tightening the skin, resisting aging, resisting wrinkles, preserving moisture and the like.

The present invention relates to the technical field of protein engineering and provides a recombinant humanized type III collagen having a triple helix structure, a preparation method therefor, and a use thereof. The amino acid sequence of the recombinant humanized type III collagen comprises n core units, wherein n is 1 or an integer greater than 1, and the amino acid sequence of the core unit is as shown in SEQ ID NO: 1. The present invention further provides a method for constructing an expression system and a method for preparing a recombinant humanized type III collagen. The present invention further provides an actual industrial use on the basis of the characteristics of the protein. The recombinant humanized type III collagen provided by the present invention has high purity, good stability, and high biological activity. Experiments have shown that the recombinant humanized type III collagen has various biological activities such as promoting cell proliferation and migration.

The invention belongs to the technical field of gene engineering and protein engineering, and particularly relates to thermal-stable D-psicose 3-epimerase as well as a preparation method and application thereof. According to the invention, a series of D-psicose 3-epimerase DAE mutants are constructed, and six mutants which not only can retain good catalytic activity, but also have high thermal stability are obtained through screening. The catalytic temperature of the three mutants A19D / A223P / V241I is increased by 25 DEG C compared with that of a wild type, while the three mutants A19D / A223P / V241I still show the optimal thermal stability, the catalytic activity of the three mutants is improved by 17.0%, and the three mutants show good industrial suitability. The D-psicose 3-epimerase DAE mutant with improved thermal stability provided by the invention is beneficial to improving the production efficiency of the DAE in industrial application of D-psicose, and provides powerful industrial enzyme guarantee for large-scale and low-cost preparation of D-psicose.

The invention relates to the technical field of functional gene and protein engineering, and discloses an amino acid sequence, a gene sequence, an expression vector and application of medicago ruthenica MrAGL8 protein, the amino acid sequence of the medicago ruthenica MrAGL8 protein is as shown in SEQ ID NO.1, the gene sequence for coding the medicago ruthenica MrAGL8 protein is as shown in SEQ ID NO.2, and the gene sequence for coding the medicago ruthenica MrAGL8 protein is as shown in SEQ ID NO.1. The invention further discloses application of the medicago ruthenica MrAGL8 protein to construction of an early-maturing plant strain or a low-pod-cracking-rate plant strain. Experiments show that the MrAGL8 gene plays an important role in promoting the early maturing of plants and reducing the pod cracking rate, and by utilizing the phenomenon that the found MrAGL8 gene participates in the response reaction of plant growth and development and pod cracking, the MrAGL8 gene is used for constructing an early maturing plant strain or a low pod cracking rate plant strain and carrying out heterologous overexpression; therefore, early maturing of transgenic plants is effectively promoted, and the pod cracking rate is remarkably reduced.

The invention provides a recombinant I-type collagen with a thermostable triple helix structure and application thereof, and belongs to the technical field of protein engineering, the amino acid sequence of the recombinant I-type collagen is shown as SEQ ID NO: 1; the invention also provides a construction method of the expression system and a preparation method of the recombinant I-type collagen. The invention also provides practical industrial application according to the characteristics of the protein.

The invention belongs to the technical direction of protein engineering in the technical field of biology, and particularly relates to a triple-helix folding and stabilizing method for recombinant collagen, which is a technology for efficiently assisting correct folding of human recombinant collagen expressed by escherichia coli and forming a stable triple-helix structure. The technology mainly solves the common key technical problems of low folding efficiency, unstable triple helix structure and the like of the recombinant collagen in an escherichia coli expression system, and obviously improves the yield of the functional recombinant collagen. The method has important application value in the fields of biomedical materials, tissue engineering, beauty and skin care and the like, and an efficient and economical technical path is provided for industrial production of recombinant collagen.

The invention provides a recombinant III-type collagen with a triple helix structure as well as a preparation method and application thereof, and relates to the field of protein engineering. According to the invention, a novel humanized III-type collagen functional fragment with biological activity is obtained through screening by selecting low-immunogenicity and high-biological activity blocks after complete sequence analysis of human III-type collagen. The functional fragments obtained by screening are repeatedly combined to obtain the novel recombinant humanized III-type collagen, and the recombinant humanized III-type collagen is a novel protein with strong biological activity and low immunogenicity, and provides a novel raw material for medical instruments and skin care products. The recombinant humanized III-type collagen sequence is efficiently expressed by adopting engineering bacteria, the expression level is high, the purification method is simple, and the recombinant humanized III-type collagen sequence is suitable for industrial production and wide popularization.

The invention relates to a chicken beta-defensin-6 mutant based on AI aided design, a recombinant expression system and application, and relates to the technical field of gene engineering and protein engineering. The chicken beta defensin-6 mutant has the following amino acids at the following five sites: lysine at the 33rd site, lysine at the 35th site, asparagine at the 49th site, arginine at the 61st site and proline at the 65th site. The chicken beta-defensin-6 mutant has high stability, activity and solubility, compared with wild chicken beta-defensin-6, the chicken beta-defensin-6 mutant has higher antibacterial activity and expression level, an efficient and safe antibacterial substitute is provided for livestock and poultry breeding, and the chicken beta-defensin-6 mutant can be widely applied to the fields of livestock and poultry breeding, animal health care and biological preservation.

The invention relates to a recombinant mussel-like mucoprotein as well as a coding gene and application thereof, and belongs to the technical field of gene engineering and protein engineering. According to the invention, partial functional sequences of different types of mussel mucin are screened, assembled and spliced to obtain a section of brand new mussel mucin-like sequence, and the sequence is optimized by escherichia coli codon preference to construct a high-expression recombinant strain. According to the recombinant strain, the soluble expression quantity of the target protein is remarkably improved by optimizing culture conditions. The recombinant protein does not carry any exogenous tag, and the provided purification method is low in cost, high in yield and suitable for large-scale production. The prepared recombinant mussel-like protein has good adhesiveness and oxidation resistance, and the modified recombinant mussel-like protein is further provided, and the biological performance of the modified recombinant mussel-like protein is better than that of unmodified recombinant mussel-like protein.

The invention discloses a methanol dehydrogenase mutant with improved catalytic activity, and belongs to the technical field of enzyme directed evolution and protein engineering. The methanol dehydrogenase is subjected to random mutation by means of directed evolution. After the construction of the plasmid library is completed, the methanol dehydrogenase with higher activity is screened by using a high-throughput and simple in-vivo screening system. Two methanol dehydrogenase mutants S1 (V37A) and S2 (I58T / M292T) with significantly improved methanol catalytic activity are obtained, V37 is subjected to saturation mutation on the basis, and mutants with improved formaldehyde generation rate are further screened. The methanol dehydrogenase mutant obtained by the invention can provide a good catalytic element for bioconversion of methanol, and has a wide application prospect.

The invention discloses a glucose-6-phosphate dehydrogenase mutant, a biological material, a catalyst and application, and belongs to the field of protein engineering, the mutant is based on a wild glucose-6-phosphate dehydrogenase amino acid sequence derived from Aspergillus nidulans FGSC A4, and has one or more amino acid residue mutations in the 111st site, the 127th site, the 214th site and the 217th site. The NADPH regeneration efficiency of the mutant disclosed by the invention is remarkably improved, and the catalytic efficiency of the preferable mutant K111L / T217L is 2.06 times that of wild glucose-6-phosphate dehydrogenase; the obtained mutant has wider reaction conditions, and has good NADPH regeneration efficiency under the conditions that the temperature is 20-50 DEG C and the pH is 6-10.

The present invention relates to the field of protein engineering, and specifically relates to a Morchella esculenta peptide and the use thereof. By means of comprehensively applying bioinformatic methods such as virtual enzymolysis, activity prediction, physicochemical property and safety evaluation, and molecular docking, one Morchella esculenta peptide having the potential effects of "inhibiting oxidation + skin whitening" is screened from Morchella esculenta, which peptide has a sequence of WWVCAK. In-vitro chemical experiments verify that WWVCAK is the most effective antioxidant and tyrosinase-inhibitory peptide. Therefore, WWVCAK can exert the effects of inhibting oxidation and skin whitening via multiple targets, multiple functions and multiple pathways. WWVCAK is expected to be developed and used in cosmetics as a functional skin-care ingredient with the twin effects of "inhibiting oxidation + skin whitening".

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.

The invention discloses a rhamnosyltransferase mutant and application thereof, and belongs to the technical field of genetic engineering. The catalytic activity of the rhamnosyltransferase is improved through a protein engineering modification method, so that the rhamnolipid yield of the pseudomonas aeruginosa is improved, reaches 62.87 g.L <-1 > and is 1.89 times that of a wild type, and the pseudomonas aeruginosa has a good industrial application prospect.

The invention discloses recombinant XVII type collagen with an anti-inflammatory effect and a preparation method of the recombinant XVII type collagen, and belongs to the technical field of protein engineering. The recombinant XVII type collagen provided by the invention comprises a plurality of repetitive units, and the repetitive units are as shown in SEQ ID NO. 1. Compared with natural XVII type collagen, the recombinant XVII type collagen is smaller in molecular weight, more stable and capable of being directly used for product production; meanwhile, the recombinant XVII type collagen disclosed by the invention can be combined with an inflammatory factor receptor to inhibit activation of a signal channel, so that the anti-inflammatory activity is improved, and therefore, the recombinant XVII type collagen disclosed by the invention can be used as a core active component to be applied to preparation of anti-inflammatory related products, and has a relatively high development prospect.

The present application relates to the field of protein engineering, in particular to a fusion protein with an expanded PAM recognition range and application thereof. The fusion protein provided by the present application comprises: a) an amino acid sequence from a non-PI domain of xCas9; b) an amino acid sequence from a PI domain of a SpCas9 variant. The fusion protein has a more relaxed PAM restriction and significantly improved editing efficiency, and is a novel editing tool with an expanded targeting range.

The invention discloses an amylase mutant. The amino acid sequence of the amylase mutant is shown as SEQ ID NO: 2 in a sequence table. The mutant is obtained by carrying out transformation and screening of protein engineering on the high-temperature alpha-amylase from thermococcus eurythermalis, and the high-temperature alpha-amylase is obtained by carrying out transformation and screening of protein engineering on the high-temperature alpha-amylase from thermococcus eurythermalis.

The present disclosure is generally related to the fields of microbial host cells, molecular biology, protein engineering, fermentation, protein production, and the like. Certain aspects of the disclosure are related to novel promoter and 5′-untranslated region a nucleic acid (DNA) sequences.

The invention relates to a binding protein B capable of binding with klebsiella pneumoniae and application of the binding protein B, and belongs to the technical field of molecular biology and protein engineering. The binding protein B provided by the invention can be specifically bound with an extramembrane part of a specific protein P24017. OMPAKLEPN of klebsiella pneumoniae, and experiments such as bimolecular fluorescence complementation and flow cytometry prove that the binding protein B has good recognition and stable binding capacity on the klebsiella pneumoniae, can be used for identifying and screening the klebsiella pneumoniae, and has good application prospects. And a key tool is provided for targeted drug development aiming at the bacterium.

The invention belongs to the technical field of gene engineering and protein engineering, and particularly relates to a fully human antagonistic antibody taking a lactylated STAT6 protein as a target spot and application of the fully human antagonistic antibody. According to the present invention, the completely humanized antagonistic antibody using the lactylated STAT6 protein as the target spot is screened from the human scFv phage antibody library, the amino acid sequence of the completely humanized antagonistic antibody is represented by SEQ ID NO: 1, and the completely humanized antagonistic antibody has the specificity of the targeted lactylated STAT6 protein, and has the application prospect in preparation of the product for diagnosis, treatment or alleviation of sepsis immunosuppression.

The present disclosure belongs relates to a 2709 alkali protease mutant modified based on molecular dynamics calculation and use thereof in liquid detergents. The dynamic structure feature of a 2709 alkali protease derived from Bacillus licheniformis are studied by using a molecular dynamics method through computer simulation, which is a unique effective method capable of studying a relationship between the structure of the 2709 alkali protease and molecular dynamics as well as an enzyme function in an atomic level. After molecular dynamics calculation, appropriate sites are selected for protein engineering to obtain the 2709 alkali protease mutant suitable for industrial application of detergents. The protease mutant has significantly improved enzyme activity under the alkaline condition, is better in stability, not only has good decontamination effect but also has good component compatibility with components in the detergent, and can be widely applied to the field of washing industry.

This invention relates to a recombinant type V collagen, its encoding gene, and its applications, belonging to the fields of genetic engineering and protein engineering technology. Based on the amino acid sequence of the α1 chain of human type V collagen, this invention utilizes bioinformatics and other methods to design the amino acid sequence, enabling the recombinant collagen to achieve higher expression efficiency and high stability while maintaining high biological activity. A novel collagen sequence containing 178 amino acids was designed. The nucleotide sequence of this recombinant collagen was optimized using E. coli codon preference to construct a high-expression recombinant strain. The recombinant collagen produced by fermentation of this strain contains no exogenous amino acids such as tags, making it a recombinant humanized type V collagen with good safety. Compared with commercially available type V collagen products, the recombinant type V collagen prepared using the above technical solution has better antioxidant activity and barrier repair capabilities, thus possessing good practical application value.

The invention discloses a urate oxidase mutant (rAfUOX) with trypsin and chymotrypsin tolerance and application of the urate oxidase mutant (rAfUOX). The invention provides a urate oxidase mutant with trypsin and chymotrypsin tolerance by modifying a key amino acid residue in a wild type aspergillus flavus urate oxidase molecule through a protein engineering technology. The activity half-life period of the recombinant aspergillus flavus urate oxidase mutant rAfUOXK49Q after trypsin digestion is increased by 244% compared with the activity half-life period of the wild type rAfUOX, and the activity half-life period of the recombinant aspergillus flavus urate oxidase mutant rAfUOXK49Q after chymotrypsin digestion is increased by 191% compared with the activity half-life period of the wild type rAfUOX. Other enzymatic properties are basically consistent with those of the wild type rAfUOX.

The invention discloses a black fungus AMY protein and a coding gene and application thereof, and belongs to the technical field of protein engineering. The invention provides a black fungus AMY protein as well as an encoding gene and application thereof in order to excavate a key enzyme gene of a starch sucrose metabolic pathway in black fungus, and discovers and verifies that the black fungus AMY protein and the encoding gene thereof play a key regulation and control role in inducing synthesis of black fungus alpha-amylase through transcriptome sequencing. A solid foundation is laid for deeply exploring the function of a key gene of a starch sucrose metabolic pathway.