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11 results about "Acetate kinase" patented technology
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In molecular biology, acetate kinase (EC 2.7.2.1), which is predominantly found in micro-organisms, facilitates the production of acetyl-CoA by phosphorylating acetate in the presence of ATP and a divalent cation. Short-chain fatty acids (SCFAs) play a major role in carbon cycle and can be utilized as a source of carbon and energy by bacteria. Salmonella typhimurium propionate kinase (StTdcD) catalyzes reversible transfer of the γ-phosphate of ATP to propionate during l-threonine degradation to propionate. Kinetic analysis revealed that StTdcD possesses broad ligand specificity and could be activated by various SCFAs (propionate>acetate≈butyrate), nucleotides (ATP≈GTP>CTP≈TTP; dATP>dGTP>dCTP) and metal ions (Mg²⁺≈Mn²⁺>Co²⁺). Inhibition of StTdcD by tricarboxylic acid (TCA) cycle intermediates such as citrate, succinate, α-ketoglutarate and malate suggests that the enzyme could be under plausible feedback regulation. Crystal structures of StTdcD bound to PO₄ (phosphate), AMP, ATP, Ap4 (adenosine tetraphosphate), GMP, GDP, GTP, CMP and CTP revealed that binding of nucleotide mainly involves hydrophobic interactions with the base moiety and could account for the broad biochemical specificity observed between the enzyme and nucleotides. Modelling and site-directed mutagenesis studies suggest Ala88 to be an important residue involved in determining the rate of catalysis with SCFA substrates. Molecular dynamics simulations on monomeric and dimeric forms of StTdcD revealed plausible open and closed states, and also suggested role for dimerization in stabilizing segment 235-290 involved in interfacial interactions and ligand binding. Observation of an ethylene glycol molecule bound sufficiently close to the γ-phosphate in StTdcD complexes with triphosphate nucleotides supports direct in-line phosphoryl transfer. The enzyme is important in the process of glycolysis, enzyme levels being increased in the presence of excess glucose. The growth of a bacterial mutant lacking acetate kinase has been shown to be inhibited by glucose, suggesting that the enzyme is involved in excretion of excess carbohydrate. A related enzyme, butyrate kinase, facilitates the formation of butyryl-CoA by phosphorylating butyrate in the presence of ATP to form butyryl phosphate.
Recombinant yeast cell
A recombinant yeast cell functionally expressing: a) a nucleic acid sequence encoding a protein comprising phospho-ketolase (PKL) activity (EC 4.1.2.9 or EC 4.1.2.22) and / or a nucleic acid sequence encoding a protein having phosphotransacetylase (PTA) activity (EC 2.3.1.8) and / or a nucleic acid sequence encoding a protein having acetate kinase (ACK) activity (EC 2.7.2.12); and / or) a nucleic acid sequence encoding a protein having transketolase activity (EC 2.2.1.1), wherein the expression of the nucleic acid sequence encoding the protein having transketolase activity is under control of a promoter (the “TKL promoter”), which TKL promoter has an anaerobic / aerobic expression ratio for the transketolase of (2) or more.
Owner:DANISCO US INC
Beta-alanine producing strain as well as construction method and application thereof
InactiveCN120888478ABacteriaAntibody mimetics/scaffoldsPhosphoenolpyruvate carboxylasePantothenic acid
The invention provides a beta-alanine producing strain as well as a construction method and application thereof. According to the bacterial strain, an acetaldehyde dehydrogenase gene adhE, a lactic dehydrogenase gene ldhA, an acetokinase gene ackA, a pyruvate dehydrogenase gene poxB, a DNA binding transcription inhibition factor coding gene lacI, an aspartate kinase thrA, a pantothenic acid synthase gene panC, an alanine synthesis transaminase gene cycA and an alanine synthesis transaminase gene yfbQ are knocked out from an E.coli W3110 genome, and the bacterial strain is obtained. A phosphoenolpyruvate carboxylase gene ppc, a pyridine nucleotide transhydrogenase coding gene pntAB, an aspartic acid transaminase gene aspC, an aspartic acid decarboxylase gene panD derived from pseudomonas aeruginosa and a beta-alanine transporter gene NCgl0580 derived from corynebacterium glutamicum are integrated at the same time, and the bacillus subtilis is used for producing beta-alanine and has the advantage of high fermentation yield.
Owner:TIANJIN UNIV OF SCI & TECH
Method for enhancing synthesis of myo-inositol as well as engineering bacteria and application thereof
The invention belongs to the field of metabolic engineering, and discloses a genetically engineered bacterium for producing myo-inositol as well as a construction method and application of the genetically engineered bacterium. The genetically engineered bacterium takes escherichia coli as an original strain, and is obtained by performing the following gene editing on a genome of the escherichia coli: non-expressed lactose operon repressor protein, glucose phosphate dehydrogenase, acetokinase, glucose phosphate isomerase and glycerol repressor protein; and overexpressing inositol-1-phosphate synthase, inositol monophosphate, glucokinase, glucose permease and glycerol kinase. Compared with the prior art, the genetically engineered bacterium disclosed by the invention has remarkable advantages in the aspects of key enzyme expression rate, metabolism specificity, fermentation period and the like, and an efficient, stable, economical and feasible solution is provided for industrial production of myo-inositol.
Owner:TIANJIN UNIV OF SCI & TECH
Construction method of escherichia coli mutant for producing succinic acid by fermentation of synthetic culture medium
The invention discloses a construction method of an escherichia coli mutant for producing succinic acid by fermentation of a synthetic medium. The method comprises the following steps: firstly, knocking out a lactic dehydrogenase gene ldhA, a pyruvate formate lyase gene pflB, a ptsG gene responsible for encoding a phosphotransferase system EIIBC protein, an ethanol dehydrogenase gene adhE, an acetokinase-phosphate transacetylase gene ackA-pta, and a ptsG gene responsible for encoding a phosphotransferase system EIIBC protein in escherichia coli; a phosphoenolpyruvate carboxykinase gene pck from bacillus subtilis is integrated at an SS9 safety site of a strain to obtain escherichia coli ESC6 with high succinic acid yield; and mutating one or more loci in one or more genes of a glucose-transcriptional inhibition factor gene mlc, a nitrate response regulatory factor gene narL and a cyclic adenylate receptor protein gene crp to obtain the escherichia coli mutant capable of producing succinic acid by fermentation of a synthetic culture medium, wherein the one or more loci in one or more genes of the glucose-transcriptional inhibition factor gene mlc, the nitrate response regulatory factor gene narL and the cyclic adenylate receptor protein gene crp are mutated. The strain can be fermented in a synthetic medium to produce succinic acid, so that the fermentation cost is greatly reduced, and the strain has a very wide application prospect.
Owner:DALIAN UNIV OF TECH
Method for detecting concentration of acetic acid in anaerobic fermentation system by using bioelectrochemical sensor
The invention discloses a method for detecting the concentration of acetic acid in an anaerobic fermentation system by using a bioelectrochemical sensor, acetic acid is detected by using the bioelectrochemical sensor, the bioelectrochemical sensor comprises a screen-printed electrode, and the working surface of the screen-printed electrode is modified with reduced graphene oxide and polymethylene blue. The working surface of the screen printing electrode is connected with acetokinase, pyruvate kinase and lactic dehydrogenase through a cross-linking agent. According to the method, the adopted sensor has better conductivity, higher response speed and higher current intensity, when the sensor is used for detecting the concentration of acetic acid in a fermentation system, the sensor has the advantages of being easy to operate, low in sample consumption, low in price, high in accurate measurement precision, high in detection speed and the like, and rapid detection of the concentration of acetic acid can be achieved; the acetic acid concentration in the anaerobic fermentation process in different periods can be timely and accurately obtained, effective data can be provided for anaerobic fermentation analysis, and the method has important significance for monitoring the process stability of anaerobic fermentation.
Owner:HUNAN UNIV
Engineered Acetylkinase Variant
This disclosure relates to engineered acetate kinase and compositions thereof, recombinant polynucleotides encoding said engineered acetate kinase, and methods of using said engineered acetate kinase.
Owner:CODEXIS INC
Method for driving biochar and nanometer magnetite to synergistically strengthen anaerobic digestion of silage feed waste under weak magnetic field
PendingCN122128369AElectrical/wave energy microorganism treatmentFermentationElectron transferBiochar
This invention discloses a method for synergistically enhancing the anaerobic digestion of silage waste using biochar and nano-magnetite driven by a weak magnetic field. This method, through the combined application of a weak magnetic field, nano-ferric oxide, and biochar, increases methane production by 67.7% during the anaerobic digestion of silage waste, enhances acetic acid production, and accelerates the conversion of acetic acid to methane. Under the drive of the weak magnetic field, the conductive network formed by biochar and nano-ferric oxide synergistically reduces the charge transfer impedance of the system, enhances the Fe(III) / Fe(II) redox cycle and interspecies direct electron transfer, and simultaneously increases the activity of key enzymes such as acetate kinase. This promotes the formation of a tighter metabolic interaction network between bacteria and archaea, ultimately achieving efficient coupling of carbon, electron, and energy flows within the anaerobic digestion system. This provides a promising technical strategy for improving the resource utilization efficiency of organic waste.
Owner:GUIZHOU UNIV
Increasing production of acetyl coa and derivative products in yeast
The present disclosure provides a recombinant yeast host cell having a variety of native and / or heterologous enzymes that function in an engineered metabolic pathway for the conversion of fructose-6-phosphate to acetyl-CoA. The recombinant yeast host cells may be used, for example, in methods for preparing acetyl-CoA, acetone and / or isopropanol. The plurality of natural and / or heterologous enzymes are activated, upregulated or overexpressed. The plurality of natural and / or heterologous enzymes comprise phosphoketolase; and optionally an acetate kinase; and / or a phosphate transacetylase. The recombinant yeast host cell comprises at least one of: at least two copies of a heterologous nucleic acid molecule encoding the phosphoketolase; a native and / or heterologous enzyme acting in the engineered non-oxidizing pentose phosphate pathway, wherein the native and / or heterologous enzyme is activated, upregulated or overexpressed; and / or a native and / or heterologous protein acting in an engineered metabolic pathway for converting pantothenic acid to acetyl-CoA, wherein the native and / or heterologous protein is activated, upregulated or overexpressed and comprises at least one of FEN2 or CAB1.
Owner:DANSTAR FERMENT AG +1
High-temperature-resistant uracil nucleotide kinase mutant and application thereof
The invention provides a high-temperature-resistant uracil nucleotide kinase mutant and application thereof, a coding gene of the mutant, and application of the mutant in preparing uridine triphosphate from uracil nucleotide through enzyme catalysis. The method for preparing uridine triphosphate from the high-temperature-resistant UMP kinase mutant has the advantages of high yield, simple process, short period, convenience in industrial production and the like. The UMP kinase and the mutant thereof can be efficiently expressed in escherichia coli, the optimum enzyme activity temperature of the mutant enzyme is 70 DEG C, and the mutant enzyme shows relatively high tolerance to a high-temperature environment of 50-70 DEG C. Under the conditions that the reaction temperature is 50 DEG C, the substrate concentration is 150 mM and acetic acid kinase is added as a coenzyme, the conversion rate of uridine nucleotide catalyzed by enzyme liquid obtained by carrying out heat treatment on the UMP kinase mutant at 70 DEG C reaches 96% within only 2 hours, so that a good technical support is provided for industrial large-scale production of uridine triphosphate.
Owner:JINGJIANG SUXING BIOTECHNOLOGY CO LTD
Adenosine kinase mutant with double-enzyme activity and application of adenosine kinase mutant
The invention provides an adenosine kinase ADKpt mutant with double-enzyme activity, a coding gene of the adenosine kinase ADKpt mutant, and application of the adenosine kinase ADKpt mutant in preparing adenosine triphosphate by efficiently catalyzing adenosine. The mutant is obtained through three-point directional combination mutation on the basis of an adenosine kinase wild type sequence, and has remarkably improved catalytic activity and good thermal stability. The ADKpt and the mutant thereof can be efficiently expressed in escherichia coli, the optimum enzyme activity temperature of the mutant enzyme is 80 DEG C, and the mutant enzyme shows relatively high tolerance to a high-temperature environment of 50-80 DEG C. Under the conditions that the reaction temperature is 60 DEG C, the substrate concentration is 200mM and acetokinase (Acek) is added as a coenzyme, the conversion rate of catalyzing adenosine within only 5 hours reaches 100% by using an enzyme solution obtained by carrying out heat treatment on the ADKpt mutant at 80 DEG C, so that a solid technical foundation is laid for industrial large-scale production of adenosine triphosphate.
Owner:JINGJIANG SUXING BIOTECHNOLOGY CO LTD
Immobilized enzyme composite material for preparing uridine triphosphate and application thereof
The invention provides an immobilized enzyme composite material for preparing uridine triphosphate (UTP) and application of the immobilized enzyme composite material. According to the invention, a uracil nucleotide kinase (UMP kinase) mutant and acetokinase (Acek) are co-immobilized on a magnetic mesoporous silica carrier through a tannic acid-metal ion (TA-Fe < 3 + >) coating technology, and the high-performance immobilized enzyme composite material is prepared. According to the material, spatial co-localization of UMP kinase and acetokinase is achieved, the ATP coenzyme regeneration efficiency is greatly improved, and the hydrolysis loss of ATP in the reaction process is effectively reduced through the physical barrier effect of the TA-Fe < 3 + > coating. In addition, the immobilized enzyme has good thermal stability and operation stability, and can be quickly recycled and reused by utilizing magnetism. The method is simple in process, low in cost, high in product purity and yield, and especially suitable for industrial large-scale continuous production of uridine triphosphate.
Owner:SUZHOU ZHENTAI BIOTECHNOLOGY CO LTD