40 results about "Hydrogenase" patented technology

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 relates to the field of biological engineering and technology, in particular to an enzyme for synthesizing glabridin, a biological synthesis method of glabridin and a bacterial strain. The invention discloses a series of glabridin synthesizing enzymes (phenylalanine ammonialyase, cinnamic acid-4-hydrogenase, tyrosine ammonialyase, p-coumaric acid coenzyme A ligase, chalcone synthase, chalcone reductase, chalcone isomerase, isoflavone synthase, 2-hydroxyisoflavone dehydratase, 4 '-oxymethyltransferase, 4'-hydroxymethyltransferase and 4 '-hydroxymethyltransferase) derived from glycyrrhiza glabra for the first time. ) can be used in the preparation of a compound (e.g., isoflavone 2 '-hydrogenase, isoflavone reductase, Vissisoketoreductase, isoprenyltransferase, pterocarpin reductase, oxidative cyclase, demethylase, cytochrome P450 reductase, and the like). The de novo synthesis of the glabridin in a microbial cell factory is realized for the first time, and the application prospect of the glabridin biological manufacturing industry is greatly promoted.

The invention provides a hydrogen production reaction system taking formaldehyde and water as co-substrates, which comprises a buffer solution, maltodextrin, formaldehyde, magnesium chloride, manganese chloride, NAD (Nicotinamide Adenine Dinucleotide), benzyl viologen, sodium phosphate, alpha-glucan phosphorylase, phosphoglucomutase, glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, 6-phosphoglucolactonase, diaphorase and hydrogenase. The enzyme is prepared from 1, 6-hexanediol phosphate synthase, hexanediol phosphate isomerase, glucose phosphate isomerase and 2, 6-hexanediol phosphate synthase. The invention also provides a corresponding hydrogen production method. Formaldehyde and water are utilized to generate hydrogen, so that excessive dependence on agricultural grain resources is avoided; meanwhile, high-energy formaldehyde is used for replacing traditional carbohydrates as a substrate, so that the energy barrier of co-substrate water is reduced, and the process of small molecular utilization of carbohydrates and hydrogen production is simplified; in addition, the method has the advantages of being not harsh in water quality condition and low in cost.

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.

The present invention discloses a method for constructing giant organelles in Escherichia coli to enhance the oxygen resistance of [Fe-Fe] hydrogenase, and belongs to the field of enzyme catalysis technology. The method comprises the following steps: constructing two plasmids capable of expressing [Fe-Fe] hydrogenase and guide protein and [Fe-Fe] hydrogenase maturation-promoting protein in Escherichia coli; transforming the two plasmids into Escherichia coli BL21 (DE3) competent cells and performing pre-culture; transferring the pre-culture to a new culture medium, performing aerobic culture, and after culturing for a period of time, determining its OD600 value, transferring the culture to an anaerobic glove box, anaerobically culturing, centrifuging, and obtaining Escherichia coli containing giant organelles. The present invention provides a method for constructing giant organelles in Escherichia coli to enhance the oxygen resistance of [Fe-Fe] hydrogenase, which effectively enhances the oxygen resistance and stability of [Fe-Fe] hydrogenase by constructing giant organelles in Escherichia coli, and provides a new sustainable method for the production of hydrogen.

The present invention belongs to the field of biochemical engineering, and relates to a chemical catalyst–microbial carbon sequestration system for improving the utilization efficiency of H2 and the use thereof. By culturing a chemical catalyst having H2 activation performance and acetogens in a culture medium containing H2 and CO2, the utilization efficiency of H2 and CO2 can be significantly improved, and the concentrations of ethanol and acetic acid are significantly increased, thereby reducing the acid-alcohol ratio; moreover, the biomass is also significantly improved. The provided hydrogenase-mimicking chemical catalyst and acetogens carbon sequestration system significantly improves the utilization efficiency of H2 and the carbon sequestration efficiency, increases the content of ethanol and thus increases the added value of the product, and is of great significance to the industrial implementation of biological fixation of CO2 using H2 as an energy source.

The invention discloses construction of a shewanella hydrogengens-CdS nanoparticle hybrid system based on a hydArsp gene and an application of the shewanella hydrogengens-CdS nanoparticle hybrid system. The method comprises the following steps: connecting a hydArsp gene derived from Rhodobacter sphaeroids coded hydrogenase to a vector pYYDT, carrying out ribosome optimization transformation to obtain a plasmid pYYDT-hydArsp, introducing the recombinant plasmid pYYDT-hydArsp into wild shewanella oneidensis MR-1 to obtain a recombinant shewanella engineering bacterium SERS with high hydrogen production efficiency, anchoring CdS nanoparticles on periplasm and an extracellular membrane of a strain in a self-assembly manner, and carrying out high-yield hydrogen production on the recombinant shewanella engineering bacterium SERS. The CdS / SERS high-hydrogen-yield active hybrid system is obtained. The CdS / SERS 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.

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-fixing 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.

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 invention relates to the fields of coordination chemistry and metal organic chemistry, and discloses a single iron hydrogenase model compound containing 3, 4 and 5-substituted pyridine ligands, the chemical structural formula is as follows: # imgabs0 #, R1 and R2 are identical or different monodentate ligands or R1 and R2 represent bidentate ligands, and R3 and R5 are identical or different and represent H or C1-C6 alkyl. R4 represents H or hydroxyl. The recombinase obtained by recombining the single iron hydrogenase model compound and decoenzyme has better stability and higher catalytic activity.

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 present invention belongs to the technical field of optimization of microbial hydrogen production, and particularly relates to a Clostridium pasteurianum biological / abiological hydrogen production system based on a hydrogen production mechanism. In this system, the endogenous hydrogenase gene of Clostridium pasteurianum is overexpressed in Clostridium pasteurianum, or a hydrogen production element is added during the cultivation of Clostridium pasteurianum, and the hydrogen production element includes one or more of Fe<supgt;2+< / supgt>, Ni<supgt;2+< / supgt>, riboflavin, humic acid or magnetic nanoparticles. The present invention explores the effects of different types of hydrogenases on hydrogen production by Clostridium pasteurianum, and clarifies the hydrogen production mechanism of Clostridium pasteurianum; by externally adding a new hydrogen production element, the hydrogen production yield of Clostridium pasteurianum is further increased, which can promote the development of biological hydrogen production technology and provide a feasible biological solution for the production of clean energy.

The invention discloses Klebsiella bacteria capable of eliminating hydrogen production capacity and application of the Klebsiella bacteria. The invention relates to a Klebsiella engineering bacterium. The engineering bacterium is a Klebsiella bacterium in which hydrogenase-3 is inactivated. The engineering bacterium provided by the invention is obtained by inactivating hydrogenase-3 of Klebsiella bacteria, cannot produce hydrogen through aerobic fermentation or anaerobic fermentation, and can effectively promote a carbon source to be converted into polyhydric alcohols. When the engineering bacterium provided by the invention is used for preparing the polyol, the advantages of high product production rate, high substrate conversion rate and the like are achieved, and the engineering bacterium has good application value and economic benefit.

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 present invention provides a hydrogenase and an enzyme catalytic system and their application in the catalytic synthesis of tetrahydromagnolol. Based on the advantages of strong specificity, high catalytic efficiency and no pollution shown by the biological method, after using the hydrogenase of SEQ ID NO.1 in the reaction of catalyzing the synthesis of 5,5'-dipropyl-[1,1'-biphenyl]-2,2'-diol with 5',5-di-2-propenyl-1,1'-biphenyl-2,2'-diphenol as the substrate, the characteristics of high conversion rate and good purity are achieved.

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.

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.

The invention relates to a chemical catalyst-microorganism carbon sequestration system for improving H2 utilization efficiency and application thereof, and belongs to the field of biochemical engineering. According to the method, the chemical catalyst with H2 activation performance and the acetogenic bacteria are cultured in the culture medium containing H2 and CO2, so that the utilization efficiency of H2 and CO2 can be remarkably improved, the concentration of ethanol and acetic acid can be remarkably improved, the acid-alcohol ratio is reduced, and the biomass is also remarkably improved. According to the hydrogenase-like chemical catalyst and the acetogenic bacterium carbon sequestration system provided by the invention, the H2 utilization and carbon sequestration efficiency are remarkably improved, and the ethanol content is increased, so that the added value of a product is increased, and the hydrogenase-like chemical catalyst and the acetogenic bacterium carbon sequestration system have important significance on industrial implementation of CO2 biological immobilization taking H2 as an energy source, and have wide application scenes and economic values.

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.