25 results about "Hydrogenase" patented technology

The application discloses a non-metal modified hydrogen evolution material and a preparation method. The Sx-CoPTA is prepared by a ligand regulation strategy, a precursor solution is prepared from cobalt nitrate hexahydrate, 4,4-biphenyldicarboxylic acid, 1,4-benzenedithiol, N,N-dimethylformamide, ethanol and water, and during the preparation, ultrasonic treatment, stirring, washing, drying and low-temperature centrifugal reaction treatment are carried out, and finally, the S-CoPTA is generated, and the hydrogen evolution electrode material is synthesized by smearing the Nafion solution on the nickel foam. The catalytic electrode material S1-CoPTA prepared by the application has a large specific surface area of the MOF material, breaks the original symmetrical structure of the MOF material, effectively improves the conductivity of the MOF material, and further improves the hydrogen evolution efficiency of the electrode material, and has a center structure and a function simulation similar to the [Fe-Fe] hydrogenase.

Methods and systems are disclosed for using industrial waste for the production of hydrogen gas. The method includes examining a pH level of the industrial waste, removing contaminate from the industrial waste, conditioning and concentrating the industrial waste to a proton-rich solution, and using the resulting proton-rich solution as the proton source in a hydrogenase catalyzed hydrogen production system.

The present invention provides a genetically modified host cell comprising a heterologous hydrogenase, or dehydrogenase, or homologous variant thereof, wherein the genetically modified host cell has an increased cellular reducing power compared an unmodified host cell. In some embodiments, the hydrogenase is a soluble hydrogenase (SH) from Cupriavidus necator H16, or a homologous variant thereof.

Methods and a recombinant C1- fixing microorganism for the continuous production of products from gaseous substrates. Further, the gaseous substrate comprises CO2 and an energy source. The recombinant C1-fxing microorganism has a disruptive mutation in a membrane bound hydrogenase gene. The C1-fixing microorganism having the disruptive mutation minimizes the H2:CO2 uptake ratio.

This invention provides a hydrogen production reaction system using formaldehyde and water as co-substrates, comprising a buffer solution, maltodextrin, formaldehyde, magnesium chloride, manganese chloride, NAD, benzyl viologen, sodium phosphate, α-glucan phosphorylase, phosphogluconomutase, glucose-6-phosphate dehydrogenase, phosphogluconate dehydrogenase, phosphogluconolactonease, flavoxase, hydrogenase, 6-phosphate hexulose synthase, phosphogluconose isomerase, and phosphogluconose isomerase. This invention also provides a corresponding hydrogen production method. This invention utilizes formaldehyde and water to generate hydrogen, avoiding excessive dependence on agricultural resources; simultaneously, by replacing traditional carbohydrates with high-energy formaldehyde as the substrate, it lowers the energy barrier of the co-substrate water and simplifies the process of carbohydrate miniaturization and hydrogen production; furthermore, it has the advantages of being less demanding in terms of water quality conditions and having low cost.

The application discloses a Talaromyces with multi-oxidase synergistic expression capacity and a fermentation method and application thereof. Talaromyces sp. The Talaromyces (Talaromyces autumnalis) Ta-lac is preserved in the China General Microbiological Culture Collection Center on July 11, 2025, and the preservation number is CGMCC No. 42125. Compared with the engineering strain for biosynthesis of laccase, the strain has strong environmental adaptability, can naturally synthesize laccase, and has relatively stable genome, no potential risk of exogenous genes and relatively high safety. The Talaromyces has a short growth cycle, high yield of laccase, nitropropane dioxygenase, hydrogenase coenzyme dioxygenase and squalene monooxygenase and high lignin degradation activity in a fermentation process. The Talaromyces Ta-lac is applied to degradation of toxin dye wastewater, and has remarkable detoxification and decolorization effects.

The present disclosure relates to a microorganism of the genus Corynebacterium, into which an exogenous soluble pyridine nucleotide transhydrogenase is introduced, and a method for producing L-tryptophan using the same.

The application provides an expression vector combination of hydrogenase, a recombinant bacterium, an enzyme composition and application thereof, and belongs to the technical field of genetic engineering and enzyme engineering. Through the combined use of formic acid dehydrogenase, formaldehyde dehydrogenase, formaldehyde enzyme and glycerol dehydrogenase, one-carbon compounds are reduced to the value-added chemical glycerol, and the rapid regeneration and efficient supply of energy cofactor NADH are realized by the biological enzyme (hydrogenase SH) method, so that the reduction equivalent is provided for the cascade catalytic reaction. In the enzyme cascade catalytic process, formic acid and methanol have strong inhibitory effect on the enzyme, the condensation of formaldehyde catalyzed by the formaldehyde enzyme can obtain neutral dihydroxy acetone, and the reaction is irreversible, and the fixation of CO2 is effectively pulled. The method guides the construction of an efficient carbon dioxide multi-enzyme immobilization system, and provides a new method and new idea for realizing carbon dioxide fixation and biological energy synthesis.

The invention discloses construction of a shewanella hydrogengens-CdS nano particle hybrid system based on a hydAson gene and an application of the shewanella hydrogengens-CdS nano particle hybrid system. The preparation method comprises the following steps: connecting a hydAson gene from Shewanella oneidensis coded hydrogenase to a vector pYYDT, carrying out ribosome optimization transformation on the vector pYYDT to obtain a plasmid pYYDT-hydAson, introducing the recombinant plasmid pYYDT-hydAson into wild shewanella oneidensis MR-1 to obtain a recombinant shewanella engineering bacterium SES with high hydrogen production efficiency, and carrying out self-assembly to obtain a CdS / SES high hydrogen production activity hybrid system. The CdS / SES hybrid system constructed by the invention endows shewanella with light capturing ability, CdS nanoparticles provide photoelectrons for engineering bacteria to promote intracellular reducing power and ATP regeneration, and the hybrid system constructed by the invention can enhance the proton reduction rate of the engineering bacteria and improve the hydrogen production efficiency.

The invention discloses a nitrogen limitation-based clostridium pasteurianum-rhodobacter sphaeroides coupling fermentation hydrogen production method and application thereof. The method comprises the following steps: performing dark fermentation on activated clostridium pasteurianum in a nitrogen-free culture medium in an argon environment to produce hydrogen; and adjusting the pH value of the obtained culture solution to 7.0-8.0, selectively sterilizing, inoculating activated rhodobacter sphaeroides, and carrying out light fermentation in an argon environment to produce hydrogen. The clostridium pasteurianum fermentation liquor can be directly subjected to rhodobacter sphaeroides photofermentation hydrogen production culture only by simply adjusting the pH value without operations such as centrifugal removal of thalli. According to the method, excessive proliferation of thalli is inhibited through nitrogen limitation, the flow direction of a carbon source is saved, high expression of nitrogenase is induced, and a second efficient hydrogen production way except hydrogenase is opened up, so that the hydrogen yield is further increased while the cost of a nitrogen source is reduced. The hydrogen production yield reaches 9.81 mol H2 / mol glucose, which is the highest level at present.

The application discloses a method for producing hydrogen through Clostridium pasteurianum-Rhodobacter sphaeroides coupling fermentation based on nitrogen limitation and application thereof. In the method, activated Clostridium pasteurianum is subjected to dark fermentation for producing hydrogen in a nitrogen-free culture medium and in an argon environment; then, the pH value of the obtained culture solution is adjusted to 7.0-8.0, the activated Rhodobacter sphaeroides is inoculated after selective sterilization, and the light fermentation for producing hydrogen is carried out in the argon environment. In the application, the fermentation solution of Clostridium pasteurianum does not need to be subjected to centrifugal removal of bacterial bodies and the like, and only needs to be simply adjusted in pH value, so that the light fermentation for producing hydrogen of Rhodobacter sphaeroides can be directly carried out. In the application, the nitrogen limitation is used to inhibit excessive proliferation of bacterial bodies, save carbon source flow, induce high expression of nitrogen fixation enzyme, and open a second high-efficiency hydrogen production path in addition to hydrogenase, so that the hydrogen production yield is further improved while the cost of nitrogen source is reduced. The hydrogen production yield of the application reaches 9.81 mol H2 / mol glucose, which is the highest level at present.

The invention discloses talaromyces sp. With a polyoxidase synergistic expression capability as well as a fermentation method and an application of the talaromyces sp. The talaromyces sp. Ta-lac is preserved in the China General Microbiological Culture Collection Center on July 11, 2025, and the preservation number of the talaromyces sp. Ta-lac is CGMCC (China General Microbiological Culture Collection Center) No.42125. The talaromyces sp. Ta-lac is named Ta-lac. Compared with an engineering strain for biologically synthesizing laccase, the strain has relatively high environmental adaptability, laccase synthesis can be naturally carried out, a genome is relatively stable, potential risks of exogenous genes do not exist, and the safety is relatively high. The talaromyces sp. Is short in growth cycle, high in yield of laccase, nitropropane dioxygenase, hydrogenase coenzyme dioxygenase and squalene monooxygenase in the fermentation process, and high in lignin degradation activity. The talaromyces talarosus Ta-lac is applied to degradation of toxin dye wastewater, and the toxicity reduction and decolorization effects are remarkable.

The invention discloses an anaerobic ammonia oxidation efficient denitrification method based on the synergistic effect of hydrazine synthase and hydrazine dehydrogenase, and belongs to the technical field of wastewater biological treatment. The method specifically comprises the following steps: providing an SBR denitrification reactor; the SBR denitrification reactor is filled with the gel beads of the immobilized double enzymes, and wastewater is introduced for operation; the double-enzyme immobilized gel bead comprises an inner core and a shell, mesoporous silica with hydrazine synthase crosslinked and immobilized in a pore channel is used as the inner core, and the shell is a hydrazine dehydrogenase layer wrapping the surface of the inner core. Precise control of a denitrification path is achieved through the first-stage reaction area and the second-stage reaction area which are physically separated, the substrate transfer efficiency is enhanced by means of the nano confinement effect, the reaction process is precisely regulated and controlled through membrane separation, finally, the total nitrogen removal rate exceeds 95%, the carrier retention rate is higher than 99%, the sludge yield is remarkably reduced (reduced by 55%), and the method is suitable for large-scale industrial production. The method is suitable for low-carbon treatment of high-ammonia-nitrogen industrial wastewater, and has high efficiency, stability and engineering applicability.

The invention belongs to the technical field of hydrogenase biology. The invention provides a fusion hydrogenase. The fusion hydrogenase contains subunits HoxF, HoxU-HhyS and HhyL, wherein the subunits are HoxF, HoxU-HhyS and HhyL; the amino acid sequence of the subunit HoxF is as shown in SEQ ID NO. 1 in a sequence table; the amino acid sequence of the subunit HoxU-HhyS is as shown in SEQ ID NO. 3 in the sequence table; the amino acid sequence of the subunit HhyL is as shown in SEQ ID NO.5 in the sequence table. The fusion hydrogenase provided by the invention can utilize low-concentration hydrogen and even hydrogen in air to regenerate NADH (Nicotinamide Adenine Dinucleotide Horse) so as to reduce the risk of operation; the NADH regeneration method has the advantages that the NADH regeneration method is simple and convenient to operate, has tolerance to oxygen and is difficult to inactivate under the aerobic condition (still has the activity of regenerating NADH), so that the application range is wider, and the operation is simpler and more convenient; nAD + can be combined more easily, and the capacity of regenerating reducing power is higher.

Provided herein, in some embodiments, are engineered cells and use of the same for increased hydrogen production. In particular, provided herein are genetically engineered cells comprising a polynucleotide encoding a fusion protein comprising a photosystem I (PSI) protein and an algal hydrogenase, as well as methods for producing such genetically engineered cells. Also provided herein are methods for increasing hydrogen (H2) production in cells.

Described herein are genelight cultures and extracts and methods of making and using thereof. In one aspect, the method of making a genelight culture or extract includes the steps of (a) making a DNA construct containing genes for producing a heat shock protein, RuBisCO large subunit 1, tonB, hydrogenase, and a P-type ATPase, (b) introducing the DNA construct into host microbial cells via transformation or transfection, and (c) culturing the microbial cells to produce the genelight cultures and extracts. The compositions of these cultures and extracts can be tailored to have specific properties such as the ability to provide power to a light emitting diode. The cultures and extracts have further uses including enhancing the growth of plants and as supplemental nutrients of cultures of industrially important microorganisms. The cultures and extracts further have UV-protective properties. Also described herein are microbial electric circuits containing the cultures and extracts described herein.

This invention discloses a method for improving the soluble expression of the large subunit of [NiFe] hydrogenase in *Escherichia coli*, belonging to the fields of protein engineering and industrial microbiology. This method involves fusing a short-chain solubilizing peptide XXA to the C-terminus of the target [NiFe] hydrogenase large subunit to form a fusion protein. This design significantly promotes the soluble expression of the target protein while preserving the accessibility of the C-terminal hydrolysis site, enabling it to transform from an inclusion body form to a soluble form under anaerobic high-density fermentation conditions. This invention overcomes the long-standing technical bias in the field that C-terminal fusion hinders the maturation and processing of [NiFe] hydrogenase, achieving a balance between solubilizing effect and processing compatibility. It provides a new technical pathway for the recombinant production of complex metalloenzymes and has significant application value in the industrial preparation of hydrogenase, basic enzymology research, and bioenergy.

A method for enhancing hydrogen uptake by cells of an acetogenic bacterium in a culture is provided. The method comprises incubating the cells in a liquid medium supplemented with an external gas mixture, wherein the external gas mixture comprises H2 and CO2; overexpressing an uptake hydrogenase and an associated electron transfer protein in the cells; increasing uptake of the H2 by the cells; and enhancing uptake of the CO2 by the cells. The culture may be a coculture with additional cells of a non-acetogenic bacterium, which produces natively H2 from catabolism of an organic molecule.

The invention discloses a clostridium aerovorans recombinant strain, a starting strain of the recombinant strain is clostridium aerovorans, and the recombinant strain is prepared by overexpressing hydrogenase CLJUc37220 in the starting strain. The hydrogenase CLJUc37220 has different hydrogen uptake rates and cofactor dependence compared with electron disproportionated hydrogenase, the expression level of the hydrogenase CLJUc37220 is improved, and the hydrogen uptake rate and the generated reducing power composition can be changed, so that the recombinant strain shows higher carbon immobilization capability.

The present application belongs to the technical field of hydrogen conversion and energy storage, and particularly relates to a method for expressing multi-subunit hydrogenase and producing formic acid by using formate dehydrogenase. The present application uses a pBBR1MCS-1 vector, adds a Lac promoter in front of each subunit sequence of hydrogenase, so that each gene is controlled by a single promoter, and adds a His tag to each gene for subsequent purification of expressed protein. After that, the constructed vector is transformed into Ralstonia eutropha H16 competent cells to obtain an engineering bacterium and express hydrogenase in the engineering bacterium. The recombinantly expressed hydrogenase is combined with formate dehydrogenase to realize self-circulation regeneration of NAD + / NADH in the reaction system without additional NADH, thereby efficiently converting H2 and CO2 into formic acid in a green and economical manner, and realizing resource utilization of green hydrogen and high-value conversion of greenhouse gas CO2.

The application discloses an engineering bacterium for synthesizing PHA by using carbon dioxide and a construction method and application thereof. First, the Rubisco gene from Methanosaeta sp. GP6 is integrated on the genome, and the original promoter is replaced; then, the hydrogenase gene from Citrobacter freundii ATCC 29220 is integrated on the genome, the lactate dehydrogenase gene in the competitive pathway is knocked out, and the PHA synthesis gene is overexpressed, so as to further improve the PHA yield. The engineering bacterium provided by the application adopts a high-density fermentation culture method, the PHA yield in a 5L fermenter can reach 83.5% of the cell dry weight, and the cell dry weight reaches 22.14g / L. The application has important application significance for industrialized production of PHA.

The invention provides a recombinant strain for producing [Fe-Fe] hydrogenase as well as a preparation method and application of the recombinant strain. The recombinant strain provided by the invention uses model bacteria escherichia coli as an expression host, the preparation method is simple, and the obtained hydrogenase has higher activity and can be further used in the field of research of biological energy.

The invention relates to a biological energy source and carbon emission reduction technology, and aims to provide a method for promoting methane sarcina to reduce CO2 by using H2 through improved coenzyme F420. Comprising the following steps: gradually domesticating methane sarcina in an air atmosphere which contains H2 and has gradually increased concentration gradient, so that the activity of coenzyme F420 in a finally obtained improved strain is improved to 3-5 times of that of an original strain, and the expression quantities of three key genes frhA, frhB and frhD of the coenzyme F420 are respectively improved by several times; the improved strain capable of stably inheriting is subjected to enlarged culture and is used for reducing CO2 by H2 to generate methane. The coenzyme F420 is directionally improved, the electron transfer rate is remarkably enhanced, the synergistic effect of a methyl coenzyme M reductase pathway and a hydrogenase system is directly promoted, the CO2 conversion efficiency and the methane production rate are improved, and the metabolic bottleneck of natural strains is broken through. By improving the strain, the activities of hydrogenase and ATP synthetase are enhanced, the cell growth division is accelerated, the apoptosis rate is reduced, and meanwhile, high metabolic activity is still maintained under low H2 partial pressure.

The invention discloses a polypeptide self-assembled nickel-selenium artificial hydrogenase as well as a preparation method and application thereof. The artificial hydrogenase is a polypeptide-nickel compound with a binuclear coordination center, wherein the polypeptide-nickel compound is formed by a self-assembly reaction of polypeptide and nickel ions in a buffer solution; a main chain of the polypeptide provides two strongly coordinated main chain amide groups, polypeptide cysteine and selenocysteine residues provide two side chain mercaptan and / or selenol, and the two side chain mercaptan and / or selenol are coordinated with Ni < 2 + > together to form a binuclear coordination center. The stability of the artificial hydrogenase metal coordination structure is improved, a binuclear metal center can be stably constructed, disordered polypeptide aggregation is reduced, the structural repeatability is improved, the electron transfer path of the metal center is more controllable, and an active structure can be kept under conventional solution and air conditions. An artificial biomimetic catalytic system with a clear structure, good repeatability and higher applicability can be obtained.

The application provides a hydrogenase gene hycE and application thereof, and belongs to the technical field of genetic engineering. The nucleotide sequence of the hydrogenase gene hycE provided by the application is shown as SEQ ID NO. 1. The hydrogenase gene hycE provided by the application is transformed into E. coli for expression, and the hydrogen production capacity and hydrogen production efficiency of the E. coli can be significantly improved. The method for producing hydrogen provided by the application has higher safety. Moreover, compared with other hydrogen production microorganisms, the E. coli has low culture cost, high realizability and controllable conditions, and is beneficial to industrial production.