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11 results about "Adenosine kinase" patented technology
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Adenosine kinase (AdK; EC 2.7.1.20) is an enzyme that catalyzes the transfer of gamma-phosphate from Adenosine triphosphate (ATP) to adenosine (Ado) leading to formation of Adenosine monophosphate (AMP). In addition to its well-studied role in controlling the cellular concentration of Ado, AdK also plays an important role in the maintenance of methylation reactions. All S-adenosylmethionine-dependent transmethylation reactions in cells lead to production of S-adenosylhomocysteine (SAH), which is cleaved by SAH hydrolase into Ado and homocysteine. The failure to efficiently remove these end products (Ado removed by phosphorylation by AdK) can result in buildup of SAH, which is a potent inhibitor of all transmethylation reactions. The disruption of AdK gene (-/-) in mice causes neonatal hepatic steatosis, a fatal condition characterized by rapid microvesicular fat infiltration, leading to early postnatal death. The liver was the main organ affected in these animals and in it the levels of adenine nucleotides were decreased, while those of SAH were elevated. Recently, missense mutations in the AdK gene in humans which result in AdK deficiency have also been shown to cause hypermethioninemia, encephalopathy and abnormal liver function.
Ergothioneine-producing recombinant engineering bacterium as well as construction method and application thereof
The invention discloses an ergothioneine-producing recombinant engineering bacterium as well as a construction method and application thereof, and belongs to the technical field of gene recombination fermentation. The method comprises the following steps: by taking escherichia coli as a starting bacterium, carrying out recombinant expression on a histidine trimethyl inner salt cysteine sulfoxide synthetase egt1 gene, an L-histidine methyltransferase egtD gene, a histidine trimethyl inner salt cysteine sulfoxide lyase egt2 gene, a methionine adenosine transferase metK gene, an adenylate kinase adK gene and an adenine phosphoribose transferase apt gene; on this basis, supply of precursor substances including histidine, cysteine and methionine is further improved through metabolic transformation, the yield of the finally obtained recombinant engineering bacterium for producing ergothioneine reaches 2.54 g / L after 48 h of shake-flask culture, the yield of a 5L fermentation tank reaches 18 g / L after further enlarged culture, the yield of ergothioneine is guaranteed while energy consumption and cost are reduced, and the method is suitable for industrial production. The method is suitable for practical popularization.
Owner:SHANGHAI RECOM BIOTECHNOLOGY CO LTD
Polyphosphate-rich yeast and application thereof in synthesis of adenosine triphosphate
The invention provides a yeast rich in polyphosphate and application of the yeast in synthesis of adenosine triphosphate, and the content of polyphosphate is 15-35 wt% based on the dry matter mass of the yeast rich in polyphosphate; according to the method disclosed by the invention, the yeast is subjected to phosphorus starvation culture and then is subjected to phosphorus supplementary culture, so that the yeast is enriched with polyphosphates, and the yeast rich in polyphosphates is obtained; then, the yeast rich in polyphosphate is used as a biocatalyst to synthesize ATP, that is, self-related enzyme systems, mainly including adenosine kinase, endo-polyphosphate kinase and excision-polyphosphate kinase, in the yeast rich in polyphosphate are used for replacing exogenous polyphosphate kinase and adenosine kinase, and in addition, ATP is synthesized by using the yeast rich in polyphosphate as a biocatalyst. According to the method for synthesizing ATP by using the yeast rich in polyphosphate as the biocatalyst, exogenous adenosine kinase and exogenous polyphosphate kinase do not need to be additionally added, the cost is low, the efficiency of converting adenosine into ATP is high, the batch-to-batch stability of the yeast is realized, and the enzyme activity of the self-related enzyme system is high.
Owner:ANGEL YEAST CO LTD +1
Method for catalytically synthesizing adenosine triphosphate by microcrystalline cellulose carrier purification immobilized enzyme
The invention discloses a method for catalytically synthesizing adenosine triphosphate by using microcrystalline cellulose carrier purification immobilized enzyme, which comprises the following steps: preparing bacterial suspension by using escherichia coli wet thalli and Tris-HCL buffer solution, crushing, centrifuging, and taking supernate to obtain crude enzyme; adding a microcrystalline cellulose carrier and the wet thalli into the crude enzyme liquid, and washing the immobilized enzyme after immobilization to obtain the immobilized enzyme; the method comprises the following steps: adding immobilized enzyme adenosine kinase and immobilized polyphosphate kinase into a substrate system with the volume of 1L according to a weight ratio, and starting a catalytic reaction under the conditions that the pH is 5.5-7.5 and the temperature is 20-40 DEG C; after catalysis is finished, the immobilized enzyme adenosine kinase and the immobilized polyphosphate kinase are subjected to solid-liquid separation with reaction liquid through a Buchner funnel, the immobilized enzyme continues to be subjected to catalytic reaction, and the method for preparing the immobilized enzyme is simple in mode, low in cost, easy to industrially produce, high in stability and capable of being repeatedly used for many times, and products are easy to separate.
Owner:ANHUI RUIBANG BIOLOGICAL SCI & TECH CO LTD
Cells, and method for producing methyl compound using cells
The purpose of the present invention is to provide a novel method whereby a methyl compound can be produced by efficiently regenerating S-adenosylmethionine (SAM) and promoting a methylation reaction, through the use of general organic raw materials such as glycine, serine, formic acid, methanol, or glucose. The present invention provides a method for producing a methyl compound using cells that have been modified so as to enhance the activity or expression of SAM-dependent methyltransferase, S-adenosylhomocysteine hydrolase (SahH), and adenosine kinase (ADO1) and reduce the activity or expression of adenylate cyclase (CyaA) and / or S-adenosylmethionine decarboxylase (SpeD).
Owner:MITSUBISHI CHEM CORP
The invention discloses a high-yield 3apos; genetic engineering strain of-deoxyadenosine and application thereof
The invention discloses a gene engineering strain with high yield of 3 '-deoxyadenosine and application of the gene engineering strain, and belongs to the technical field of gene engineering. The genetic engineering strain is obtained by carrying out genetic modification on the strain, and the genetic modification comprises at least one of the following steps: a) overexpressing a ribonucleotide reductase gene rnr1; b) inactivating / weakening an adenosine kinase gene ad1; c) inactivating / weakening the adenine deaminase gene aah1; d) overexpressing a ribonuclease gene rny1; and e) an inactivation / weakening inhibition type vacuolar alkaline phosphatase gene pho8. According to the method, substrate metabolism kinetics is accurately matched in combination with an intermittent feeding strategy, finally, the yield of 3 '-deoxyadenosine reaches 20.58 g / L and reaches the highest yield reported at present, a'gene-process' dual-drive technical barrier is formed, and an efficient, stable and low-cost solution is provided for industrial production of nucleoside compounds.
Owner:NANJING TECH UNIV
Combination of culture medium components
The present disclosure provides a culture medium for cell culturing, the culture medium containing a combination of specific components. The present disclosure provides a culture medium which contains a combination of components selected from the group consisting of: (1) a halcinonide component; (2) an ALK4 / 7 inhibitor or a TGFβ receptor type-1 kinase inhibitor; (3) a BRD7 / 9 double degradation factor or a BRD7 / 9 binding molecule (degradation factor); (4) an LATS1 / 2 inhibitor, a PKA inhibitor, or an AKT1 inhibitor; (5) a CK1 / 2 inhibitor, an insulin receptor inhibitor, a Tyr kinase inhibitor, a PKA inhibitor or a PKC inhibitor, or an adenosine kinase inhibitor; (6) an E-selectin inhibitor, a VCAM1 inhibitor, or an ICAM1 inhibitor; and (7) an LATS1 inhibitor + an LATS2 inhibitor.
Owner:SUMITOMO CHEM CO LTD +1
Halophilic adenosine kinase mutant as well as coding gene and application thereof
The invention discloses a halophilic adenosine kinase mutant as well as a coding gene and application thereof. According to the present invention, the adenosine kinase (EC 2.7. 1.20) with the amino acid sequence represented by SEQ ID NO: 1 and derived from the Candida parapolymorpha (ATCC26012) is subjected to the rational modification, such that the halophilic adenosine kinase mutant of the present invention has high catalytic activity and high salt tolerance compared to the wild adenosine kinase;
Owner:CHANGZHOU UNIV
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
Recombinant engineering strain for producing ergothioneine and construction method and application thereof
This invention discloses a recombinant engineered bacterium producing ergothioneine, its construction method, and its application, belonging to the field of gene recombination fermentation technology. This invention uses *Escherichia coli* as the starting bacterium to recombinantly express histidine trimethylammonium cysteine sulfoxide synthase. egt1 Gene, L-histidine methyltransferase egtD Gene, histidine trimethyl inner salt cysteine sulfoxide lyase egt2 Gene, methionine adenosine transferase metK Gene, adenosine kinase adK Genes and adenine phosphoribosyltransferase apt The gene was modified, and further metabolic modification was used to increase the supply of precursor substances histidine, cysteine, and methionine. The resulting recombinant engineered ergothionein-producing bacteria achieved a yield of 2.54 g / L after 48 h of shake-flask culture, and a yield of 18 g / L after further expansion to a 5L fermenter. This method reduces energy consumption and costs while ensuring ergothionein production, making it suitable for practical application.
Owner:SHANGHAI RECOM BIOTECHNOLOGY CO LTD
Recombinant expression vector, genetically engineered bacterium and application of genetically engineered bacterium in synthesis of 3 '-phosphoadenosine-5'-phosphate sulfate
The invention discloses a recombinant expression vector, a genetically engineered bacterium and application of the genetically engineered bacterium in synthesis of 3 '-adenosine-5'-phosphate sulfate, and belongs to the field of bioengineering and enzyme engineering. According to the method, a multi-enzyme system containing adenosine kinase (atADK) and polyphosphate kinase (chPPK2) is constructed, adenosine and polyphosphate (PolyP) which are low in price are used as starting raw materials, efficient regeneration of ATP is achieved, and PAPS is catalytically synthesized by coupling pyrophosphatase (PPA), adenosine 5 '-phosphosulfate kinase (APSK) and ATP sulfating enzyme (KAST). The constructed genetically engineered bacterium can be carried out in an in-vitro one-pot method or a whole-cell system, a good conversion rate is achieved, the conversion rate is larger than 50%, and the yield can reach 5.12 mM PAPS. The biological production cost can be reduced, and meanwhile, the method has industrial application prospects.
Owner:杭州裕元生物科技有限公司 +1