15 results about "Hydrogenase" patented technology

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 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 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.

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 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.