70 results about "Carboxysome" patented technology

The present application relates to a mutant gene encoding phosphoenolpyruvate carboxylase containing a mutant 5' untranslated region (5'UTR), a polyhydroxyalkanoate (PHA)-producing microorganism containing the gene, and a PHA production method using the same.

The present disclosure relates to a genetically modified microorganism satisfying some of predetermined conditions. The predetermined conditions include: (I) succinate dehydrogenase activity or fumarate reductase activity being reduced or inactivated relative to a wild-type microorganism; (II) lactate dehydrogenase activity being reduced or inactivated relative to the wild-type microorganism; (III) the genetically modified microorganism having modified phosphoenolpyruvate carboxylase activity showing resistance to feedback inhibition by aspartic acid in wild-type phosphoenolpyruvate carboxylase activity, or exogenous phosphoenolpyruvate carboxylase activity having higher resistance to feedback inhibition by aspartic acid than that of the wild-type phosphoenolpyruvate carboxylase activity shown by the wild-type microorganism; and (IV) pyruvate:quinone oxidoreductase being reduced or inactivated relative to the wild-type microorganism.

The application provides a recombinant Yarrowia lipolytica capable of producing palmitoleic acid at a high yield, a construction method and application thereof, and belongs to the technical field of bioengineering. Yarrowia lipolytica The recombinant Yarrowia lipolytica is obtained by knocking out peroxisome biogenesis factor 10 and triacylglycerol lipase 4 in the genome of Yarrowia lipolytica and inserting an acetyl-CoA carboxylase, a Δ9 desaturase, an acetyl-CoA diacylglycerol acyltransferase, a glycerol-3-phosphate acyltransferase, a lysophosphatidylcholine acyltransferase and a phosphatidylcholine:diacylglycerol acyltransferase gene expression cassette. Experiment proves that the recombinant Yarrowia lipolytica can efficiently ferment and produce palmitoleic acid, and realizes efficient synthesis of the natural product palmitoleic acid from plants in Yarrowia lipolytica.

The application belongs to the field of biological analysis, and provides a method for precise metabolic flow analysis of TCA cycle based on isotope tracing technology combined with high-resolution mass spectrometry. By optimizing mass spectrometry parameters and liquid chromatography method, one injection within 6 min is realized, and high-throughput, high-sensitivity and high-resolution detection is realized on 11 biomarkers related to TCA cycle metabolic pathways. At the same time, the application innovatively uses LC-MS / MS to decipher 13 C the stepwise position-specific transfer from glucose to subsequent metabolites by glycolysis and TCA cycle, thereby calculating 13 C the integrated steady-state analysis of key metabolic rates of position-specific transfer from sequential precursors to their products (pyruvate dehydrogenase, beta-oxidation, pyruvate carboxylase, isocitrate dehydrogenase and pyruvate cycle). This technology has wide applicability and can potentially characterize mitochondrial metabolism of any tissue or cell.

The present application relates to the field of plant genetic breeding, and particularly relates to a kind of evaluation method of asparagus yield. Including the following steps: S1, obtaining young plant period asparagus;S2, measures i variety of young plant period asparagus following parameters: photosynthetic rate A, ribulose-1,5-bisphosphate carboxylase / oxygenase activity B, leaf dry weight per unit area C, acid invertase activity D and root crown ratio E;The measurement sample of the acid invertase activity D is the tender stem of young plant period asparagus at 10-30cm from top;S3, according to the formula, the evaluation value is calculated, and the evaluation value is used to evaluate the yield of asparagus. The evaluation value obtained by using the above measurement index and calculation formula, and the significance level obtained by SPSS software is used to evaluate the yield of asparagus. The yield potential of asparagus can be comprehensively evaluated from the genetic point of view in a short time. Using the evaluation method of the present application, high-yield asparagus varieties can be selected in a short time without tracking for many years.

This invention aims to create a spatially isolated local reaction microenvironment for Rubisco by introducing a complete β-carboxysome microcompartment system. This design goal avoids oxygen inhibition, laying the foundation for ultimately achieving efficient and specific CO2 fixation. It provides a novel approach to overcome the fundamental problem of low efficiency of carbon-fixing enzymes in cytosols due to intense oxygen competition. By introducing the complete carboxysome system as a module, engineered yeast strains with high carboxylase activity and significantly improved ethanol production were obtained, providing a solid foundation for building next-generation yeast cell factories that produce high-yield ethanol using CO2 as a raw material.

The invention discloses a recombinant bacterium for producing mevalonic acid by taking glucose and acetone as co-substrates as well as a construction method and application of the recombinant bacterium, and belongs to the technical field of genetic engineering. The problems that an existing mevalonic acid biosynthesis mode is low in carbon atom economy, limited in theoretical yield and the like are solved. According to the invention, acetone carboxylase, acetoacetyl coenzyme A synthetase, 3-hydroxy-3-methylglutaryl coenzyme A reductase and hydroxymethyl glutaryl-CoA synthetase are subjected to heterologous expression in escherichia coli with an acetyl coenzyme A acetyltransferase gene atoB and a histidine protein kinase gene atoS knocked out, and self carbonic anhydrase is over-expressed; and a new way for synthesizing MVA by using glucose and acetone as a co-substrate is constructed. According to the method, one molecule of CO2 can be fixed in the biosynthesis of mevalonic acid, the yield of carbon atoms in the synthesis process of mevalonic acid is increased, the problem of low yield of carbon atoms in the synthesis process of mevalonic acid by taking glucose as a single substrate is solved, and a new thought is provided for constructing a carbon neutralization type biological manufacturing technology.

The invention discloses a compound enzyme preparation-containing nutritional feed for regulating and controlling intramuscular fat of black pigs and a preparation method of the compound enzyme preparation-containing nutritional feed, and relates to the technical field of livestock and poultry breeding feeds. The invention discloses a black pig skeletal muscle cell fat directional transport feed which is prepared from the following components in parts by weight: 850-920 parts of basal feed, 5-12 parts of fat directional deposition compound enzyme group, 25-40 parts of amino acid-enzymatic synthesis substrate combination and 2-6 parts of lipolytic enzyme specific inhibitor. According to the black pig skeletal muscle cell fat directional transport feed, fat precursor directional transport is realized through the compounded fat directional deposition compound enzyme group, a fat synthesis pathway targeting black pig skeletal muscle cells and lipoprotein lipase; acetyl-coenzyme A carboxylase and fatty acid synthetase catalyze fat synthesis step by step, only intramuscular fat generation pathways are activated, subcutaneous and abdominal fat deposition paths are blocked, the technical defects that fat deposition of an existing single enzyme preparation is non-directional, and the body fat rate is caused to be abnormally increased are overcome, meanwhile, an exclusive amino acid-enzymatic synthesis substrate combination is matched, and the body fat synthesis efficiency is improved. Sufficient adaptive substrates are supplied to enzyme group catalytic reaction, and the problem that a single enzyme preparation is insufficient in catalytic efficiency is solved.

This invention provides an engineered bacterium for producing ectoine, its preparation method, and its application. The engineered bacterium is prepared using a method comprising the following steps: using *Corynebacterium glutamicum* as the starting strain, feedback inhibition by aspartate kinase LysC is relieved, the activity of the gene lysE encoding lysine efflux permease is reduced, and the ectoine synthesis gene cluster ectABC is expressed. Based on this, the activity of any one or at least two of the following—diaminobutyrate acetyltransferase EctA, pyruvate carboxylase Pyc, or aspartate kinase LysC—is enhanced, and / or the activity of any one or at least two of the following—maloquinone oxidoreductase Mqo or homoserine dehydrogenase Hom—is reduced to further increase ectoine yield.

Disclosed are genetically engineered Schizosaccharomyces pombe strains capable of fermenting glucose to produce malic acid. The S. pombe strains produce an exogenous malate dehydrogenase (MDH) enzyme and optionally produce an exogenous pyruvate carboxylase (PYC) enzyme or an exogenous phosphoenolpyruvate carboxylase (PPC) enzyme. Additionally, the S. pombe strains may be further engineered to produce a fumarase selectively located in mitochondria and / or have an inactivated mitochondrial citrate transporter, so that the resulting S. pombe strains are capable of producing malic acid as the predominant C4 dicarboxylic acid. Also disclosed are methods of producing malic acid with the S. pombe strains.

The application belongs to the technical field of biology, and discloses a strain for efficiently fixing carbon dioxide and improving succinic acid conversion rate and a construction method thereof. The application discloses a genetically engineered bacterium, which is obtained by overexpressing carbonic anhydrase genes, phosphoenolpyruvate carboxylase genes and phosphoenolpyruvate carboxykinase genes in a succinic acid-producing strain. ‑ The carbonic anhydrase can enhance the conversion efficiency of CO2 to HCO3 , and the phosphoenolpyruvate carboxylase and the phosphoenolpyruvate carboxykinase can realize efficient fixation of C3 and CO2 into C4 intermediates. Experiments prove that the genetically engineered bacterium can effectively produce succinic acid by using glucose, and has a relatively high sugar acid conversion rate (up to 79.80%).

The composition of the present invention is suitable for use in preparing emulsion-type meat analogue, preferably as a sausage type in a casing, which comprises rubisco protein and kappa-carrageenan in relative amounts between 1:5 to 30:1. Generally, the composition comprises 0.5-30 wt % rubisco and 0.1-15 wt % carrageenan relative the finished meat analogue product, and vegan protein other than rubisco preferably in an amount of 0.5-30 wt % relative to the meat analogue product. The meat analogue may contain the composition described in an amount of between 2-40 wt % with 30-90 wt % water and 2-40 wt % plant-based lipid.

The application discloses genetically engineered bacteria for producing alpha-ketoglutaric acid and application. The genetically engineered bacteria are obtained by weakening the expression of alpha-ketoglutaric acid dehydrogenase gene odhA, weakening the expression of glutamate transporter protein gene cg1434, weakening the expression of glutamate synthase key subunit gene gltB, weakening the expression of lactic acid dehydrogenase gene ldh, weakening the expression of glutamate dehydrogenase gene gdh, weakening the expression of serine / threonine protein kinase gene pknG, enhancing the expression of pyruvate carboxylase gene pyc, introducing and expressing glutamate oxidase gene lgox at the site of lactic acid dehydrogenase ldh gene, and enhancing the expression of strong catalase gene katA on the basis of a Corynebacterium glutamicum starting strain. The genetically engineered bacteria can efficiently produce alpha-ketoglutaric acid under the condition of limited ammonia culture, and have a good application prospect.

The application provides a pyroglutamic acid transaminase mutant and application thereof, the mutant is obtained by mutation on the basis of wild-type pyroglutamic acid transaminase, and specific mutation includes at least one of the following sites: (1) the 13th isoleucine is mutated into arginine; (2) the 35th valine is mutated into arginine; (3) the 183rd asparagine is mutated into aspartic acid; and the mutant is suitable for industrial enzymatic production of L-aspartic acid and the process of amino acids taking L-aspartic acid as a precursor; especially, the I13R / V35R / N183D mutant obtained by modification has high positive catalytic activity and strict direction selectivity, and completely loses the reverse reaction activity of decomposing L-aspartic acid; the mutant is overexpressed with phosphoenolpyruvate carboxylase in L-threonine production strain THRS-6, and a high-efficiency L-threonine synthesis metabolic pathway is constructed, and the yield of L-threonine is significantly improved when glucose is used as a substrate.

The present application relates to the technical field of genetic engineering, and particularly relates to a phosphoenolpyruvate carboxylase mutant and application thereof. The phosphoenolpyruvate carboxylase mutant is obtained by a plurality of mutations in the amino acid sequence of the phosphoenolpyruvate carboxylase. The present application provides a plurality of mutation sites of the phosphoenolpyruvate carboxylase, and further finds that after the mutation of the sites of the phosphoenolpyruvate carboxylase in the strain, the yield of amino acids such as glutamic acid, glutamine and proline is significantly improved, which has important significance in the field of improving the strain to produce glutamic acid and its derivatives.

The invention provides a genetic engineering strain for efficiently producing beta-alanine, an escherichia coli strain is modified by adopting a genetic engineering means, and a multi-plasmid system is constructed, so that a metabolic pathway is optimized and the product synthesis efficiency is improved. The method comprises the following steps: firstly, by constructing a recombinant plasmid pET-PanD, introducing aspartic acid decarboxylase (L17F / G24R mutant) with mutation so as to promote efficient conversion of aspartic acid to beta-alanine; meanwhile, phosphoenolpyruvate carboxylase (PPC) is overexpressed through the recombinant plasmid pRSF-PPC, generation of oxaloacetic acid (OAA) is increased, and then the synthesis path of carbon flow to beta-alanine is improved. Besides, NADH dependent type aspartate dehydrogenase (AspDH) from pseudomonas aeruginosa is overexpressed by adopting a pBAD-AspDH plasmid, so that the dependence on NADPH is reduced, and the conversion efficiency is further improved.

The invention discloses an artificial synthesis method and application of malonyl coenzyme A. An artificial synthesis route for synthesizing malonyl-CoA by taking beta-alanine (beta-ala) as a precursor is constructed by heterologous expression of aminotransferase and malonyl-CoA reductase: firstly, under the catalysis of transaminase, amino of beta-ala is transferred to alpha-keto acid (such as pyruvic acid, oxaloacetic acid or alpha-ketoglutaric acid and the like), and then the malonyl-CoA is obtained; an intermediate product 3-oxopropionic acid and corresponding amino acid are formed; and the malonyl-CoA is generated from the 3-oxopropionic acid under the action of the malonyl-CoA reductase. According to the method, the defects existing in a natural malonyl-CoA synthesis route, such as low carbon utilization rate, energy substance ATP consumption, greenhouse gas CO2 release, pyruvate dehydrogenase (PDH) of route enzymes and acetyl-CoA carboxylase (ACC) which are strictly regulated and controlled, are overcome, and high yield of products taking malonyl-CoA as a precursor, including light flavomycin, octanoic acid, phloroglucinol, pentadecheptaene, natamycin, spinosad and the like, is realized.

This invention provides a recombinant acetyl-CoA carboxylase encoding gene and its applications. Mutations at positions 5374, 5229, 5335, 5336, and 5493 in the recombinant acetyl-CoA carboxylase encoding gene provided by this invention can induce more significant resistance in rice expressing this recombinant acetyl-CoA carboxylase. In particular, it shows a significant increase in resistance to seven ACCase inhibitors from three major chemically distinct classes: arbutin (APP), quizalofop-P-ethyl, quizalofop-P-ethyl, and flupyrflupyr; cyclohexenone (CHD), clethodim and clodinafop-P-ethyl; and neophenylpyrazoline (DEN), clodinafop-P-ethyl.

Methods and systems are provided for the purification of native and recombinant Ribulose- 1,5- bisphosphate Carboxylase / Oxygenase (RuBisCO) from cellular lysates using liquid-liquid phase separation (LLPS). The invention enables scalable and cost-effective extraction of highly pure RuBisCO complexes from plant and bacterial sources while maintaining functional protein integrity. The methods and compositions disclosed herein address key obstacles to RuBisCO isolation and offer commercial advantages for protein production and utility in nutritional formulations.

The invention belongs to the technical field of carbon dioxide fixation. The invention provides a biological carbon sequestration enzyme mutant. The biological carbon sequestration enzyme mutant is obtained after threonine at the 398th site of RPE Rubisco is mutated; the amino acid sequence of the RPE Rubisco is as shown in SEQ ID NO. 1 in a sequence table. The threonine mutation is that threonine is mutated into alanine. According to the invention, through mutation of a single amino acid, the catalytic efficiency of Rubisco is successfully improved, and especially, the turnover number of enzyme is greatly increased and reaches 35.84 S <-1 >. Through reasonable structural design, the trade-off effect between kcat and km is successfully avoided, so that the Rubisco keeps high CO2 affinity while the activity is improved. The method is simple, is suitable for design and modification of other carboxylases, and has a wide application prospect.

The present application provides a recombinant E. coli for synthesizing heme and derivatives thereof, wherein the recombinant E. coli is knocked out of genes such as pyruvate oxidase poxB, pyruvate formate-lyase pflB and protoporphyrinogen peroxidase yfeX, and meanwhile, the genes such as phosphopyruvate carboxylase ppc, glycerol transporter glpF and 3-phosphoglycerate dehydrogenase glpD are enhanced, and the genes such as 5-aminovaleryl-CoA synthase hemA and ferrous chelatase hemH are introduced, so that the recombinant E. coli can synthesize heme and derivatives thereof by using glycerol as a raw material, and has application prospects in the field of biological manufacturing.

The invention discloses a method for synthesizing salicylic acid and derivatives thereof by multi-enzyme cascade catalysis of phenol, which comprises the following steps: reacting a reaction system containing phenol, enzyme, metal ion salt and bicarbonate to obtain salicylic acid and derivatives thereof; when the enzyme is carboxylase, the reaction system reacts to obtain salicylic acid; when the enzyme is a combination of carboxylase, carboxylate reductase, formate dehydrogenase and phosphokinase, the reaction system reacts to obtain salicylaldehyde; when the enzyme is a combination of carboxylase, carboxylate reductase, formate dehydrogenase, phosphokinase and alcohol dehydrogenase, the reaction system reacts to obtain salicyl alcohol; when the enzyme is a combination of carboxylase, carboxylate reductase, formate dehydrogenase, phosphokinase, alcohol dehydrogenase and glycosyltransferase, the reaction system reacts to obtain salicin. According to the method, a green synthesis path based on biological catalysis is utilized, phenol is converted into chemicals with higher economic value, and environmental pollution and carbon emission are reduced.

Aspects of the present disclosure relate to the discovery that antagonizing pyruvate carboxylase and glutaminase inhibits cell proliferation and extracellular matrix production. Thus, methods and compositions disclosed herein may be used to treat fibrotic disease and cancer by inhibiting cell proliferation and extracellular matrix production.

The invention discloses a construction method and application of an engineering bacterium for increasing the yield of alpha-ketoglutaric acid. Wild E.coli.W3110 serves as a chassis strain, lactic dehydrogenase ldhA, pyruvate dehydrogenase poxB and formic acid C-acetyltransferase pflB are knocked out firstly, and the flow direction of pyruvic acid to lactic acid, acetic acid and formic acid is blocked; meanwhile, the promoter PBBaJ23109 is used for replacing a natural promoter of the sucA gene, so that alpha-ketoglutaric acid dehydrogenase is weakened, and the decomposition way of alpha-ketoglutaric acid is weakened; the phosphoenolpyruvate carboxylase ppc is overexpressed, and the flow direction of phosphoenolpyruvate to oxaloacetic acid is enhanced; isocitrate dehydrogenase icd from escherichia coli is overexpressed, so that synthesis of a product is promoted; besides, pyruvate dehydrogenase derived from saccharomyces cerevisiae is introduced and coded by genes PDB1, PDA1 and LAT1, so that an important precursor, namely pyruvic acid, is promoted to flow to acetyl coenzyme A; the engineering strain with high yield of alpha-ketoglutaric acid is finally obtained, the yield of 75.1 g / L can be achieved in a 5L fermentation tank through a fed-batch fermentation method for 40 hours, and the conversion rate is 39.6%.

The present invention provides a hydroxyacetyl-CoA carboxylase (GCC) wherein said GCC is characterized in that it comprises an amino acid sequence having at least 60% sequence identity to SEQ ID NO: 1, and at a position selected from the group consisting of position 20th and position 100th in the amino acid sequence represented by SEQ ID NO: 1 or a position corresponding to any of these positions, the GCC is characterized in that the GCC has at least one amino acid sequence having at least 60% sequence identity to SEQ ID NO: 1, and the GCC has at least one amino acid sequence having at least 60% sequence identity to SEQ ID NO: 2. The present invention relates to a GCC having one or more amino acid substitutions, deletions or insertions, preferably wherein the GCC has improved activity relative to a reference GCC, preferably wherein the reference GCC comprises the amino acid sequence of SEQ ID NO: 1.

The invention discloses a construction method and application of escherichia coli genetically engineered bacteria for producing malic acid by synergistically utilizing CO2, and belongs to the technical field of bioengineering. According to the invention, an L-malic acid chassis strain MG1655W (DE3) constructed by a gene editing technology in a laboratory is used as an initial strain, a lactic dehydrogenase encoding gene ldhA and a phosphate acetyltransferase encoding gene pta in escherichia coli are knocked out on the basis, the production paths of lactic acid and formic acid are blocked, and the yield of lactic acid and formic acid is increased. The method comprises the following steps of: firstly, amplifying a phosphoenolpyruvate carboxylase gene ppc and a CO2 transporter gene CA, constructing a recombinant plasmid, introducing the recombinant plasmid pACYCDuet-1-CA-ppc into an expression host MG1655W (DE3) delta ldhA delta pta, and screening to obtain the engineering bacterium MG1655W (DE3) delta ldhA delta pta-CA-ppc by overexpressing the ppc gene and the CA gene and improving the CO2 immobilization and concentration capability of escherichia coli. The obtained engineering bacterium is subjected to cell proliferation under an aerobic condition and malic acid accumulation under an oxygen limiting condition, the final malic acid yield is increased to 140-212%, and the engineering bacterium has important industrial application value.

The invention discloses a cyhalofop-butyl-resistant digitaria sanguinalis MYB transcription factor gene and application thereof, and relates to the field of plant genetic engineering, and the expression of the gene in a resistant digitaria sanguinalis biotype is remarkably up-regulated. After a plant expression vector is constructed and corn is transformed, after 11.25 g a.i. Ha-1 cyhalofop-butyl is sprayed on a transgenic corn plant in a 3-4 leaf period, the pesticide resistance and the survival rate are obviously enhanced, and the transgenic corn plant is obviously superior to a wild type. The invention proves that the MYB gene can endow plants with cyhalofop-butyl resistance, provides a new gene resource for cultivating transgenic crops resisting acetyl-CoA carboxylase (ACCase) inhibitor herbicides, and has important application value.

This application discloses the application of the pyc gene and its mutants in the preparation of lysine, belonging to the field of genetic engineering technology. The technical problem this application aims to solve is: how to increase the amino acid yield of microorganisms. To solve the above technical problem, this application provides a protein obtained by mutating serine (S) at position 1059 of pyruvate carboxylase to proline (P) and related biological materials. This application also provides the application of said protein and related biological materials in constructing recombinant bacteria and preparing L-amino acids. This application has obtained a mutant pyruvate carboxylase (pyc) that increases the lysine yield in target microorganisms through extensive experiments. S1059P The results of the recombinant bacterial validation showed that, compared with the original target microorganism and recombinant microorganisms that overexpress wild-type pyruvate carboxylase (e.g., recombinant Corynebacterium glutamicum), the L-lysine production of recombinant Corynebacterium glutamicum expressing this mutant pyruvate carboxylase was significantly increased.

The invention provides an engineering bacterium for producing Ectoine as well as a preparation method and application thereof, and the engineering bacterium is prepared by the preparation method comprising the following steps: taking corynebacterium glutamicum as a starting strain, relieving feedback inhibition of aspartate kinase LysC, reducing activity of gene lysE for coding lysine efflux permease, and obtaining the Ectoine-producing engineering bacterium. And expressing an Ectoin synthetic gene cluster ectABC. On the basis, the activity of any one or a combination of at least two of diaminobutyric acid acetyltransferase EctA, pyruvate carboxylase Pyc or aspartate kinase LysC is enhanced, and / or the activity of any one or a combination of two of malic acid quinone oxidoreductase Mqo or homoserine dehydrogenase Hom is reduced, so that the yield of Ectoine is further improved.

The invention discloses a synergistic composition containing isoxazoline insecticidal and acaricide, which is composed of a compound I and a compound II, the compound I is CCXN-15-005-S (S configuration) or an agrochemically acceptable salt thereof, and the compound II is CCXN-15-005-S (S configuration) or an agrochemically acceptable salt thereof. The compound II is selected from any one of a nicotinic acetylcholine receptor agonist, a chordinal TRPV channel regulator, an acetyl-coenzyme A carboxylase (ACCase) inhibitor, a mitochondrial electron transfer inhibitor, a calcium-activated potassium ion channel regulator and trifluoroethyl thioether (sulfoxide) or an agrochemically acceptable salt thereof. The synergistic composition can be used for preventing and treating pests and harmful mites of thysanoptera, hemiptera and acarus and has a wider agricultural application prospect.