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33 results about "Formic dehydrogenase" patented technology

Recombinant clostridium and construction method and use thereof

The invention discloses a recombinant clostridium, a method for constructing the same and application thereof. The method for constructing the recombinant clostridium is to introduce a gene for encoding formic dehydrogenase into a clostridium to construct a recombinant bacterium. The clostridium is an acetone butanol clostridium or a clostridium beijerinckii. The invention also discloses a method for producing butanol, which is to ferment and culture the recombinant bacterium constructed by the method for constructing the recombinant clostridium to produce the butanol. The yield of butanone produced through fermenting the recombinant acetone butanol clostridium constructed by the invention can reach as high as 14.1 grams per liter.
Owner:INST OF MICROBIOLOGY - CHINESE ACAD OF SCI

Construction of coenzyme efficient regeneration system and application thereof

The invention relates to a method for preparing alpha-aminobutyric acid by utilizing series connection amino acid dehydrogenase and codon optimization formate dehydrogenase recombinant escherichia coli. First, codon optimization is performed on a Candida boidinii formate dehydrogenase gene sequence according to the codon preference of the escherichia coli. After codon optimization on formate dehydrogenase, the expression quantity of formate dehydrogenase in the escherichia coli can be improved remarkably, the yield of alpha-aminobutyric acid is increased, and in the escherichia coli, co-expression formate dehydrogenase and L-amino acid dehydrogenase can promote circulation of cofactors in mycetome without needing the adding of any exogenous cofactors. In addition, the process of utilizing whole-cell transformation bulk chemical L-threonine to produce alpha-aminobutyric acid is simple and quick, and the cost is low. In a 5 L fermentation tank, the yield of alpha-aminobutyric acid obtained through the method can be 81.5 g / L, and an actually effective strategy is provided for industrial production of alpha-aminobutyric acid.
Owner:ANHUI HUAHENG BIOTECH

Bioactive hollow nano-fibers and hollow microcapsules for efficiently catalyzing conversion of CO2 into methanol

The present invention provides bioactive hollow nano-fibers and hollow microcapsules for efficiently catalyzing methanol synthesis through CO2. According to the present invention, a coaxial co-spinning technology is used to embed three enzymes for catalyzing methanol synthesis through CO2 and comprising formic dehydrogenase, formaldehyde dehydrogenase and ethanol dehydrogenase, and coenzyme NADH into the chamber of polyelectrolyte-doped hollow nano-fibers or hollow microcapsules; in order to achieve coenzyme regeneration, an oxidation reduction enzyme R adopting NAD<+> as coenzyme is embedded into the chamber; in order to accelerate a CO2 hydration reaction, carbonic anhydrase is immobilized on the outer surface of the hollow nano-fibers or hollow microcapsules through a layer-by-layer self-assembly technology; the small molecule NADH is bound on the inner wall of the hollow nano-fibers or hollow microcapsules through the electrostatic interaction between the NADH and the polyelectrolyte in the shell layer so as to achieve the recycling; and the polyelectrolyte-doped hollow nano-fiber or hollow microcapsule CO2 conversion catalyst has advantages of simple preparation, high conversion rate, and high stability, and provides wide application prospects in the field of the conversion of CO2 into methanol.
Owner:INST OF PROCESS ENG CHINESE ACAD OF SCI

Synthesis method of chiral tert-leucine and final product obtained in method

The invention provides a synthesis method of chiral tert-leucine, and the synthesis method is characterized by comprising the following steps: adding raw materials and water to a reaction kettle; adding ammonium formate solid and adding a pH regulator to regulate the pH value after completely dissolving; then sequentially adding an ammonium formate buffer of catalyst oxidized coenzyme, a buffer solution of formic dehydrogenase and a buffer solution of chiral leucine dehydrogenase; stirring to start reaction while controlling the reaction temperature at 10-40 DEG C and the pH value of the reaction system at 6.0-10.0; and after the reaction is finished, acquiring a final product, namely deuterated chiral tertleucine. The synthesis method provided by the invention has the advantages that: the used raw materials are easily available and inexpensive; the used raw materials are commercial raw materials or easily prepared raw materials, and can meet the needs of large-scale production; and the product is obtained by one-step reaction, water is used as the solvent, enzyme protein is used as the catalyst, and the product has a yield more than 80% and both chemical purity and chiral purity greater than 99%.
Owner:ASYMCHEM LAB TIANJIN +4

Method for preparing highly pure L-tertiary leucine through biological process

The invention discloses a method for preparing highly pure L-tertiary leucine through a biosynthesis process. The method comprises the following steps: inoculating an Escherichia coli seed liquid expressing leucine dehydrogenase gene and formic dehydrogenase gene into in a fermenting culture medium of a self-induction system, carrying out fermenting culture, centrifuging to obtain crude thalli, and adding a buffer solution to obtain a cell suspension; adding substrates comprising trimethylpyruvic acid and ammonium formate, and carrying out a biological catalysis reaction to obtain an trimethylpyruvic acid conversion solution; and centrifuging the conversion solution, adjusting the pH value of the solution, carrying out column chromatography, precipitating, and concentrating to obtain highly pure L-tertiary leucine. The method fully uses the high effectiveness, the specificity and the mildness of an enzyme; and compared with traditional chemical synthesis methods, the method disclosed in the invention has the advantages of high selectivity, mild conditions, simple process, low cost, small pollution and the like.
Owner:山东斯递尔化工科技有限公司

Efficient hydrogen-production functional gene vector pET32a-fdhF as well as construction and application thereof

The invention provides an efficient hydrogen-production functional gene vector pET32a-fdhF as well as construction and application thereof and relates to the technical field of genetic engineering. The gene vector pET32a-fdhF is a pET32a plasmid containing formate dehydrogenase fdhF genes of Bacillus cereus. The construction of the gene vector comprises the steps of amplifying segments of the formate dehydrogenase fdhF genes in Bacillus cereus by virtue of a design primer, linking the segments to a pET32a segment containing a corresponding linking terminal by virtue of recombinase, carrying out gene sequencing, and verifying the integrity of the genes. The pET32a-fdhF plasmid is converted into bacteria, and the bacteria are cultured in a hydrogen-production culture medium, so as to produce hydrogen. The pET32a-fdhF can be applied to different bacteria, so that the bacteria with no hydrogen-production capacity can produce hydrogen, the hydrogen-production of the bacteria capable of producing hydrogen is obviously improved, and more hydrogen can be rapidly produced by virtue of the same substrate concentration. According to the pET32a-fdhF, existing escherichia coli for industrial hydrogen production can be improved.
Owner:BEIJING UNIV OF TECH

Prepn process of compound degrading enzyme

InactiveCN1609205AOrderly metabolismEasy to prepareEnzymesFood treatmentPhytaseCyanide
The pesticide degrading compound enzyme is prepared with lignoperoxidase, phytase, phenoloxidase, formamidase, cyanide hydrolase, formic dehydrogenase, lysozyme, and Tween solution, and through mixing, culture, drying and packing. The product has high activity, high efficiency and wide degradation spectrum, and has the functions of killing bacteria, decontaminating, eliminating pollutant heavy metal, degrading pesticide residue, etc.
Owner:陈玉柱

Method for improving resistance of formic acid and acetic acid in cellulose hydrolysate by utilizing formic acid dehydrogenase

The invention discloses a method for improving resistance of formic acid and acetic acid in cellulose hydrolysate by utilizing formic acid dehydrogenase. The method improves the tolerance capability of a strain to acetic acid and formic acid in the cellulose hydrolysate by overexpressing the formic acid dehydrogenase. In the method provided by the invention, the formic acid dehydrogenase can be overexpressed in a cellulose ethanol production strain, the resistance of the strain to formic acid and acetic acid in the cellulose hydrolysate is improved, the method is simple and feasible, the material detoxification cost is reduced, and the industrial production of the cellulose ethanol is facilitated.
Owner:DALIAN UNIV OF TECH

Method for improving conversion rate of chiral amine

The invention discloses a method for improving conversion rate of chiral amine. The method comprises the following steps: synthesizing and amplifying an optimized ArR-omega TA nucleotide sequence containing enzyme cutting sites, constructing p2A4-ArR-omega TA, p2A4-FA and p2A4-ldh, wherein FA is a gene for expressing formic acid acetyl transferase, and correspondingly introducing escherichia colito obtain recombinant bacteria 1, FA and 6; fermenting to obtain fermentation liquor of the recombinant bacteria 1, FA and 6; centrifuging to obtain fermentation liquor precipitates of the recombinantbacteria 1, FA and 6; and correspondingly preparing crude FA enzyme liquid and crude ldh enzyme liquid; resuspending the fermentation liquor precipitate of the recombinant bacteria 1 by using PBS, adding PLP and NAD< +>, and reacting; and adding the crude FA enzyme liquid, the crude ldh enzyme liquid, formate dehydrogenase, D-alanine, a substrate ketone and CoA, adding a cosolvent, reacting, terminating, extracting and drying. According to the method disclosed by the invention, a byproduct pyruvic acid is removed, and the conversion rate of the chiral amine is improved. The formate dehydrogenase is added to recover a cofactor reduced coenzyme I.
Owner:TIANJIN UNIV

Method for promoting cell-catalyzed allitol production by utilizing recombinant intracellular-RNA-support-immobilized recombinase

ActiveCN111378612ASimultaneous assembly possibleEasy to operateBacteriaBiofuelsIntracellularRecombinase
The invention discloses a method for promoting cell-catalyzed allitol production by utilizing recombinant intracellular-RNA-support-immobilized recombinase. The method comprises the steps: (1) constructing a triangular reticular RNA support for immobilizing an intracellular recombinant protein; (2) constructing recombinant plasmids for anchoring ribitol dehydrogenase and formate dehydrogenase in the triangular reticular RNA support; (3) constructing Escherichia coli engineering strains employing the triangular reticular RNA support immobilized recombinase to catalyze allitol; and (4) conducting fermentation by using the Escherichia coli engineering strains constructed in the step (3) to produce the allitol. According to the method, the ribitol dehydrogenase and the formate dehydrogenase are anchored by using the RNA support, the distance of an enzymatic reaction is shortened, and the synthesis of the allitol is efficiently promoted. Proven by experiments, the allitol yield of the engineering strains constructed by the method disclosed by the invention can be increased to 4.433g / L from 3.036g / L, so that the yield of the product is advantageously maximized in industrial practical applications.
Owner:SHANDONG UNIV +1

Fusion protein as well as coding gene and application thereof

The invention discloses a fusion protein as well as a coding gene thereof and application thereof. The fusion protein provided by the invention is protein which is obtained by in series connecting the connecting peptides of ethanol dehydrogenase and formate dehydrogenase. The ethanol dehydrogenase is from lactobacillus brevis and the formate dehydrogenase is from candida boidinii; the connecting peptides are rigid structures Linker (EAAAK). According to the experiment in the invention, the invention provides the fusion protein which is obtained by in series connecting the connecting peptides of ethanol dehydrogenase and formate dehydrogenase, wherein enzyme activity of the formate dehydrogenase is higher than the activity for independently expressing the formate dehydrogenase after the formate dehydrogenase is expressed in escherichia coli; moreover, the stability and heat stability of the ethanol dehydrogenase in organic substrate are also improved. The construction of the fusion protein provides important reference for the improvement to the preparation method for chiral alcohol and chiral hydroxy compound.
Owner:INST OF BIOENG ACAD OF MILITARY MEDICAL SCI OF THE CHINESE

Formic acid and CO2 autotrophic recombinant escherichia coli and construction method thereof

The invention discloses formic acid and CO2 autotrophic recombinant escherichia coli and a construction method thereof, and belongs to the field of microbial metabolic engineering and the field of synthetic biology. According to the invention, the most common escherichia coli is used as a starting strain, and formic acid assimilated modular plasmids and CO2 assimilated modular plasmids are introduced to construct the escherichia coli which rapidly grows by using formic acid and CO2 as a unique carbon source. The whole construction process is simple, the period is short, and the universality ishigh. According to the recombinant bacterium, the level of an intracellular reduced cofactor NAD (P) H can be improved by virtue of a formate dehydrogenase mutant, so that the recombinant bacterium has the capability of promoting pyruvic acid synthesis from beginning to end by eating formic acid, and finally formic acid and CO2 autotrophic escherichia coli is obtained; the escherichia coli is inoculated to a culture medium taking formic acid and CO2 as a unique carbon source for fermentation culture, 0.9 OD600 biomass can be obtained at most within 56 hours, and the original strain hardly grows. The escherichia coli is methanoic acid and CO2 autotrophic escherichia coli with the highest growth speed under the condition of no assistance of gene knockout, long-term domestication and other means at present.
Owner:JIANGSU UNIV

Ammonia ion diagnosis/measuring reagent kit and ammonia ion concentration determination method

The invention relates to a kit for diagnosing / measuring ammonia (ions) by utilizing the technologies of the enzymic colorimetry and the enzyme linked immunosorbent assay. The invention further relates to a method, a principle and the composition and the components of a reagent for measuring the concentration of ammonia (ions), and belongs to the technical field of medical / food / environmental inspection and measurement. The main components of the kit include a buffer solution, reduced coenzyme, glutamic acid, adenyl pyrophosphate, oxaloacetic acid, glutamine synthetase, pyruvate carboxylase, formic dehydrogenase and a stabilizer. Through mixing a sample and the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet / visible light analyzer, and the velocity of the decrease in absorbance at 340 nm of the dominant wavelength is detected, thereby measuring the concentration of ammonia (ions).
Owner:SUZHOU ANJ BIOTECHNOLOGY CO LTD

Ammonia ion diagnosis/measuring reagent kit and ammonia ion concentration determination method

The invention relates to a kit for diagnosing / measuring ammonia (ions) by utilizing the technologies of the enzymic colorimetry and the enzyme linked immunosorbent assay. The invention further relates to a method, a principle and the composition and the components of a reagent for measuring the concentration of ammonia (ions), and belongs to the technical field of medical / food / environmental inspection and measurement. The main components of the kit include a buffer solution, reduced coenzyme, glutamic acid, adenyl pyrophosphate, oxaloacetic acid, glutamine synthetase, phosphoric acid enol pyruvate carboxylase, formic dehydrogenase and a stabilizer. Through mixing a sample and the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet / visible light analyzer, and the degree / velocity of the decrease in absorbance at 340 nm of the dominant wavelength is detected, thereby measuring the concentration of ammonia (ions).
Owner:SUZHOU ANJ BIOTECHNOLOGY CO LTD

Glycine diagnosis/measuring reagent kit and glycine concentration determination method

The invention relates to a kit for diagnosing / measuring glycine by utilizing the technologies of the enzymic colorimetry and the enzyme linked immunosorbent assay. The invention further relates to a method, a principle and the composition and the components of a reagent for measuring the concentration of the glycine, and belongs to the technical field of medical / food inspection and measurement. The main components of the kit include a buffer solution, coenzyme, methyl sulfate phenazine, ferricytochrome B1, hydrogen cyanide synthetase, formic dehydrogenase, NAD dependent formic dehydrogenase and a stabilizer. Through mixing a sample and the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet / visible light analyzer, and the degree / velocity of the increase in absorbance at 340 nm of the dominant wavelength is detected, thereby measuring the active concentration of the glycine.
Owner:SUZHOU ANJ BIOTECHNOLOGY CO LTD

Kalium ion diagnosis/measuring reagent kit and kalium ion concentration determination method

The invention relates to a kit for diagnosing / measuring potassium (ions) by utilizing the technologies of the enzymic colorimetry and the enzyme linked immunosorbent assay. The invention further relates to a method, a principle and the composition and the components of a reagent for measuring the concentration of the potassium (ions), and belongs to the technical field of medical / food / environmental inspection and measurement. The main components of the kit include a buffer solution, reduced coenzyme, adenosine diphosphate, a phosphoenolpyruvic acid, coenzyme A, pyruvate kinase, pyruvate oxidase, formic dehydrogenase and a stabilizer. Through mixing a sample and the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet / visible light analyzer, and the velocity of the decrease in absorbance at 340 nm of the dominant wavelength is detected, thereby measuring the concentration of potassium (ions).
Owner:SUZHOU ANJ BIOTECHNOLOGY CO LTD

Carbon dioxide diagnosis/measuring reagent kit and carbon dioxide concentration determination method

The invention relates to a kit for diagnosing / measuring carbon dioxide by utilizing the technologies of the enzymatic cycling amplification method, the enzymic colorimetry and the enzyme linked immunosorbent assay. The invention further relates to a method, a principle and the composition and the components of a reagent for measuring the concentration of the carbon dioxide, and belongs to the technical field of medical / food / environmental inspection and measurement. The main components of the kit include a buffer solution, coenzyme, ferricytochrome b1, formic dehydrogenase (1), formic dehydrogenase (2) and a stabilizer. Through mixing a sample and the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet / visible light analyzer, and the degree / velocity of the increase in absorbance at 340 nm of the dominant wavelength is detected, thereby measuring the active concentration of the carbon dioxide.
Owner:SUZHOU ANJ BIOTECHNOLOGY CO LTD

Lithium diagnosis/measuring reagent kit and lithium concentration determination method

The invention relates to a kit for diagnosing / measuring lithium by utilizing the technologies of the enzymic colorimetry and the enzyme linked immunosorbent assay, wherein, the activity of the kit can be inhibited by lithium. The invention further relates to a method, a principle and the composition and the components of a reagent for measuring the concentration of lithium, and belongs to the technical field of medical / industrial / environmental inspection and measurement. The main components of the kit include a buffer solution, reduced coenzyme, magnesium chloride, inositol-1-phosphate, adenosine diphosphate, an oxaloacetic acid, inositol-1-phosphatase, pyruvate carboxylase, formic dehydrogenase and a stabilizer. Through mixing a check sample and a lithium sample respectively with the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet / visible light analyzer, the degree / velocity of the decrease in absorbance at 340 nm of the dominant wavelength is detected, and the difference in the degree / velocity of the decrease between the check sample and lithium sample is compared, thereby measuring the concentration of lithium.
Owner:SUZHOU ANJ BIOTECHNOLOGY CO LTD

Recombinant escherichia coli with high succinic acid yield and construction method and application thereof

ActiveCN114015634ADoes not affect growth ratePromote accumulationBacteriaHydrolasesSuccinic acidPromoter
The invention relates to the technical field of bioengineering, in particular to recombinant escherichia coli with high succinic acid yield and a construction method and application thereof. According to the recombinant escherichia coli with high succinic acid yield, a gene fdhF for encoding formate dehydrogenase in escherichia coli is replaced by a formate dehydrogenase gene fdh1 from candida, and an outer membrane protein regulator OmpR for resisting acid stress and osmotic pressure stress is up-regulated. The formate dehydrogenase gene fdhF coded in host bacteria FMME-N-5 is replaced by a formate dehydrogenase gene fdh1 of candida by adopting a CRISPR-cas9 gene editing technology, and a key outer membrane protein regulator OmpR resistant to acid stress and osmotic pressure stress is up-regulated by applying an RBS sequence and a promoter strategy; the obtained recombinant escherichia coli has no resistance, and has osmotic pressure resistance and can efficiently produce succinic acid.
Owner:JIANGNAN UNIV +1

Kalium ion diagnosis/measuring reagent kit and kalium ion concentration determination method

The invention relates to a kit for diagnosing / measuring kalium (ions) by utilizing the technologies of the enzymic colorimetry and the enzyme linked immunosorbent assay. The invention further relates to a method, a principle and the composition and the components of a reagent for measuring the concentration of kalium (ions), and belongs to the technical field of medical / food / environmental inspection and measurement. The main components of the kit include a buffer solution, reduced coenzyme, tryptophan, ferricytochrome b1, tryptophanase, pyruvate dehydrogenase, formic dehydrogenase and a stabilizer. Through mixing a sample and the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet / visible light analyzer, and the velocity of the decrease in absorbance at 340 nm of the dominant wavelength is detected, thereby measuring the concentration of kalium (ions).
Owner:SUZHOU ANJ BIOTECHNOLOGY CO LTD

Carbonyl reductase mutant and application thereof in preparation of steroid hormone-testosterone

The invention belongs to the technical field of bioengineering, and particularly relates to a carbonyl reductase mutant and application thereof in preparation of steroid hormone-testosterone. The method comprises the following steps: co-expressing a carbonyl reductase mutant PmCRm and formate dehydrogenase BstFDH_m in Escherichia coli, and catalyzing androstane-4-ene-3,17-dione to synthesize testosterone by taking resting cells of co-expressed engineering bacteria as a biocatalyst. The biocatalyst has high catalytic activity, regioselectivity and stereoselectivity, 28.8 g / L of androstane-4-ene-3,17-dione can be completely converted within 10 h to generate the target product testosterone, and no by-product is generated. The substrate feeding amount, the conversion rate and the space-time conversion rate of the catalyst all reach the highest level of testosterone prepared by a domestic biological method; after separation and purification, the product recovery rate is up to 95% or above, which indicates that the biocatalyst is an efficient catalyst for green synthesis of testosterone.
Owner:FUZHOU UNIV

A single-cell factory for synthesizing l-phenylglycine and its construction and application

The invention discloses a single-cell factory for efficiently synthesizing L-phenylglycine and its construction and application, belonging to the technical field of microorganisms. The present invention first realizes the high-efficiency expression of leucine dehydrogenase derived from Bacillus cereus in Escherichia coli, and obtains mutant N71S with significantly improved reduction properties through site-directed mutation, and combines the mutant enzyme with formate dehydrogenase mutant Co-expressed in Escherichia coli to form an intracellular in situ cofactor NADH cycle system, optimized the expression of formate dehydrogenase mutants through promoter and RBS sequence optimization, successfully constructed a recombinant Escherichia coli single-cell factory, and used the single-cell factory Carry out whole-cell transformation to prepare L-phenylglycine. The conversion process of this method is simple and fast, with low cost and no by-products, which is beneficial to separation and purification. After 4 hours of conversion in a 5L fermenter, the output of L-phenylglycine can reach 105.7g / l. The conversion rate was 93.3%, and the space-time yield of L-phenylosine was 26.3 g / L·h, which provided a practical and effective strategy for the industrial production of L-phenylosine.
Owner:JIANGNAN UNIV

Carbon dioxide diagnosis/measuring reagent kit and carbon dioxide concentration determination method

The invention relates to a kit for diagnosing / measuring carbon dioxide by utilizing the technologies of the enzymic colorimetry and the enzyme linked immunosorbent assay. The invention further relates to a method, a principle and the composition and the components of a reagent for measuring the concentration of the carbon dioxide, and belongs to the technical field of medical / food / environmental inspection and measurement. The main components of the kit include a buffer solution, reduced coenzyme, ferricytochrome b1, formic dehydrogenase, formaldehyde dehydrogenase and a stabilizer. Through mixing a sample and the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet / visible light analyzer, and the degree / velocity of the decrease in absorbance at 340 nm of the dominant wavelength is detected, thereby measuring the active concentration of the carbon dioxide.
Owner:SUZHOU ANJ BIOTECHNOLOGY CO LTD

Kalium ion diagnosis/measuring reagent kit and kalium ion concentration determination method

The invention relates to a kit for diagnosing / measuring kalium (ions) by utilizing the technologies of the enzymic colorimetry and the enzyme linked immunosorbent assay. The invention further relates to a method, a principle and the composition and the components of a reagent for measuring the concentration of kalium (ions), and belongs to the technical field of medical / food / environmental inspection and measurement. The main components of the kit include a buffer solution, reduced coenzyme, tryptophan, coenzyme A, tryptophanase, pyruvate oxidase, formic dehydrogenase and a stabilizer. Through mixing a sample and the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet / visible light analyzer, and the velocity of the decrease in absorbance at 340 nm of the dominant wavelength is detected, thereby measuring the concentration of kalium (ions).
Owner:SUZHOU ANJ BIOTECHNOLOGY CO LTD
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