1852results about "Acyltransferases" patented technology

The invention belongs to the technical field of bioengineering, and discloses a recombinant yarrowia lipolytica capable of efficiently and rapidly producing sclareol, which is characterized in that optimized sclareol synthase related coding genes ScLPPS, ScSCS, GGPPS1, GGPPS2, ERG20 and tHMGR coding genes are inserted into a yarrowia lipolytica genome; inserting optimized pentadienol utilization pathway coding genes such as IPK, THIM and IDI coding genes; and inserting an optimized ATP: citrate lyase coding gene ACLY to obtain the ATP: citrate lyase coding gene ACLY. The recombinant yarrowia lipolytica is based on knockout of a yarrowia lipolytica strain responsible for coding a non-homologous recombination gene ku70, so that the homologous recombination capability of the yarrowia lipolytica strain is enhanced, gene integration is realized through the homologous recombination function of the yarrowia lipolytica, the genetic stability of the introduced gene can be greatly improved, and the recombinant yarrowia lipolytica strain has a good application prospect. The construction method of the recombinant yarrowia lipolytica strain is efficient and simple to operate.

The present disclosure provides guided microbial remodeling (GMR) methods for the rational improvement of plant-associated microbes to perform plant-beneficial functions. The GMR methods described herein allow for non-intergeneric genetic optimization of key regulatory networks within the microbes, which improve plant-beneficial functions over wild-type microbes but don't have the risks associated with transgenic approaches (e.g., unpredictable gene function, public and regulatory concerns, etc.). The present disclosure also provides remodeled microbes and compositions thereof. The utilization of remodeled microbes and compositions thereof will enable farmers to realize more productive and predictable crop yields without the nutrient degradation, leaching, or toxic runoff associated with traditional synthetically derived fertilizers.

The invention belongs to the technical application field of microbial metabolic engineering and synthetic biology, and particularly relates to a pichia pastoris engineering strain for high-yield sclareol as well as a construction method and application of the pichia pastoris engineering strain. The method comprises the following steps: constructing a biosynthetic pathway of sclareol in a host strain, and optimizing an intracellular mevalonic acid metabolic pathway and a central metabolic pathway to obtain an engineering strain A; the host strain is pichia pastoris; or overexpressing or knocking out a metabolism regulation factor in the obtained engineering bacterium A to obtain an engineering bacterium strain B; or, in the obtained engineering bacterium B, a synthetic pathway is targeted to peroxisome and optimized by adopting a cell region compartment strategy, and an engineering bacterium strain C is obtained. The invention provides a pichia pastoris chassis cell for synthesizing a diterpenoid sclareol and an engineering strain for synthesizing sclareol. By introducing and optimizing the expression of a sclareol synthesis pathway, the fermentation yields of sclareol in shake flask batch and bioreactor batch fed-batch fermentation are respectively 631.6 mg / L and 10.5 g / L.

The invention provides a genetically engineered bacterium, a preparation method thereof and application of the genetically engineered bacterium in synthesis of sclareol. The genetically engineered bacterium comprises a CcLPPS gene derived from Cistanus crenatus, and a CcLPPS gene derived from Cistanus crenatus, the genetically engineered bacterium is selected from yarrowia lipolytica. According to the invention, the stably expressed sclareol efficient de novo synthesis strain is constructed, and no organic reagent or any path intermediate substance is added in the whole fermentation process, so that the production cost is greatly reduced, and the subsequent separation and purification steps are simplified. The engineering yarrowia lipolytica strain provided by the invention is high in production intensity and simple in fermentation process, has a relatively good application prospect, and lays a foundation for producing sclareol by a microbial fermentation method.

The present disclosure concerns a fusion polypeptide for making acetyl-phosphate from glucose, a recombinant yeast host cell expressing the fusion polypeptide as well as methods of using the fusion polypeptide. The fusion polypeptide comprising (i) a glucose-6-phosphate isomerase moiety and (ii) a phosphoketolase moiety.

The invention discloses a wheat E3 ubiquitin ligase gene RFEL1 and application of the wheat E3 ubiquitin ligase gene RFEL1 in wheat disease resistance, and belongs to the technical field of plant genetic engineering. The invention discloses a nucleotide sequence of a wheat E3 ubiquitin ligase gene RFEL1 as shown in SEQ ID NO. 3. The invention further provides application of the wheat E3 ubiquitin ligase gene RFEL1 or related biological materials thereof. The application comprises any one of the following applications: A1, improving the resistance of wheat to stripe rust; a2, the resistance of wheat to powdery mildew is improved; and A3, the resistance of wheat to leaf rust is improved. The E3 ubiquitin ligase RFEL1 gene provided by the invention not only has broad-spectrum resistance to different stripe rust, but also has resistance to wheat leaf rust and powdery mildew. The E3 ubiquitin ligase RFEL1 gene disclosed by the invention is applied to broad-spectrum resistance breeding of wheat and has important application value.

The present disclosure provides a recombinant yeast host cell having a plurality of native and / or heterologous enzymes that function in an engineered metabolic pathway to convert fructose-6-phosphate to acetyl-coenzyme A. The recombinant yeast host cell can be used, for example, in a process for making acetyl-coenzyme A, acetone and / or isopropanol. The plurality of native and / or heterologous enzymes is activated, upregulated, or overexpressed. The plurality of native and / or heterologous enzymes comprises a phosphoketolase; and optionally an acetate kinase; and / or a phosphotransacetylase. 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 that functions in an engineered non-oxidative pentose phosphate pathway, wherein the native and / or heterologous enzyme is activated, upregulated, or overexpressed; and / or a native and / or heterologous protein that functions in an engineered metabolic pathway to convert pantothenate into acetyl-coenzyme A, wherein the native and / or heterologous protein is activated, upregulated or overexpressed and comprises at least one of: FEN2 or CAB1.

The invention discloses production of terpenoids and a strain, and belongs to the technical field of bioengineering. The serratia marcescens engineering bacteria for producing the semi-terpenoids or monoterpenoids are constructed, and can be used for producing linalool, isoprene, isopentenol, 1, 3, 4-triterpenes and 1, 3, 4-triterpenes. The composition is prepared from 1, 8-cineole, alpha-pinene, pinene, gamma-terpinene, geraniol, (+)-limonene, (-)-limonene, myrcene, beta-ocimene, sabinene, (-)-alpha-bisabolol, (-)-alpha-bisabolol, longifolene, wallenenes, beta-elemene, farnesene, patchouli alcohol, acyclic terpene and alpha-santalene. In a 30L fermentation tank, the yield of linalool produced by the serratia engineering bacteria is 40.72 g.L <-1 >.

The invention relates to a nucleic acid molecule of a rice transformation event RN85zN-eH and a detection method. The nucleic acid molecule of the rice RN85zN-eH comprises a sequence shown in SEQ ID NO: 1 or a reverse complementary sequence thereof, or a sequence shown in SEQ ID NO: 2 or a reverse complementary sequence thereof. The rice transformation event RN85zN-eH disclosed by the invention has the characteristics of insect resistance and glufosinate-ammonium herbicide resistance, and the detection method can accurately and quickly identify whether a biological sample contains DNA molecules of the transgenic rice event RN85zN-eH or not.

Plants producing seeds and seeds comprising increased oil content are provided. Additionally, methods for generating and using plants and seeds having increased oil content are disclosed.

The invention relates to a recombinant pichia pastoris strain capable of producing astaxanthin at high yield as well as a construction method and application of the recombinant pichia pastoris strain. The recombinant pichia pastoris strain is obtained by expressing ATP (adenosine triphosphate) citrate lyase (ACL) and acetyl-CoA synthase (ACS) or phosphoketolase (PK) and phosphotransacetylase (PTA), choline kinase (CK), inositol polyphosphate kinase (IPK) and vitreoscilla hemoglobin (VHB) in host bacteria. Wherein the host bacterium is a pichia pastoris gene modified strain PP-LC2. The astaxanthin production performance of the recombinant strain is verified on the basis of comparison of precursor supply pathways, IUP pathway construction and improvement of the oxygen supply capacity of the engineering strain, and the astaxanthin production capacity of the pichia pastoris is further improved. The construction method of the recombinant pichia pastoris is simple, the synthesis of astaxanthin can be better promoted, the engineering strain can efficiently synthesize the astaxanthin by utilizing methanol through amplification fermentation of a 5L fermentation tank, and industrial production is facilitated.

The invention relates to the technical field of dye synthesis, and particularly discloses a genetically engineered bacterium for synthesizing N-acetyl blue as well as a construction method and application of the genetically engineered bacterium. The supply capacity of precursor substances glutamic acid and glutamine of the genetically engineered bacterium is high, the fermentation process is simple, the production intensity of the precursor substances glutamine and N-acetylglutamine is improved by cutting off a consumption path, relieving negative feedback inhibition and enhancing the expression level of key genes through the genetically engineered bacterium, and finally the yield of N-acetylblue reaches 41.2 g / L.

A nucleic acid molecule comprising: i) a first nucleic acid comprising a double stranded inhibitory ribonucleic acid (RNA) molecule comprising a sense and an antisense strand designed with reference to a nucleotide sequence comprising a cardiovascular disease associated gene to be silenced and wherein said cardiovascular gene is proprotein convertase subtilisin kexin 9 (PCSK9) there is provided a single stranded deoxyribonucleic acid (DNA) molecule conjugated to the 5’ end of said sense strand; and ii) a second nucleic acid comprising a double stranded inhibitory ribonucleic acid (RNA) molecule comprising a sense and an antisense strand designed with reference to a different cardiovascular disease associated gene to be silenced wherein said cardiovascular gene is lipoprotein (a) (Lp(a)) and wherein there is provided a single stranded deoxyribonucleic acid (DNA) molecule conjugated to the 5’ end of said sense strand wherein the single stranded deoxyribonucleic acid (DNA) molecule is substantially complementary to the single stranded deoxyribonucleic acid (DNA) molecule set forth in i) above and anneals by complementary base pairing to form a double stranded DNA linker that links the first and second double stranded inhibitory ribonucleic acid (RNA) molecules.

The present disclosure provides a recombinant yeast host cell for converting a carbohydrate to ethanol and acetone. The recombinant yeast host cell comprises an engineered metabolic pathway to convert acetyl-CoA to acetone, a downregulated native alcohol dehydrogenase (ADH) gene, and a heterologous alcohol dehydrogenase.

The invention discloses a Sarcabomyces lipolytica strain for synthesizing (2S)-naringenin and application of the Sarcabomyces lipolytica strain, and belongs to the technical field of engineering and bioengineering. According to the invention, a key enzyme CHS is subjected to semi-rational transformation, so that the yield of the strain (2S)-naringenin expressing the mutant V196A / T132C is increased by 60%. According to the method, the modified CHS is further integrated into a yeast genome, conversion of p-coumaric acid to (2S)-naringenin is promoted by strengthening a shikimic acid pathway and enhancing supply of cofactors, so that the yield of the (2S)-naringenin reaches 776mg / L after the constructed strain is fermented in a 24-pore plate for 96 hours, and the yield of the constructed strain can reach 8.65 g / L after the strain is fermented in a 5L fermentation tank; the method lays a foundation for the subsequent biosynthesis of flavonoid compounds, and has potential value and significance for the development of synthetic biology.

Provided herein, inter alia, is an engineered non-plant cell that produces a hyoscyamine alkaloid product, a precursor of a hyoscyamine alkaloid product, or a derivative of a hyoscyamine alkaloid product. Also described is a method for producing a scopolamine alkaloid, a precursor of a scopolamine alkaloid product, or a derivative of a scopolamine alkaloid product using a cell.

The invention discloses corynebacterium glutamicum for producing N-acetylglucosamine and application of the corynebacterium glutamicum, and belongs to the technical field of synthetic biology. Specifically, the corynebacterium glutamicum overexpresses a GNA1 gene from a brewer's yeast S288C controlled by a tacM promoter, a glmS gene from corynebacterium glutamicum S9114 and a yqab gene from escherichia coli K12 MG1655, and simultaneously has five gene defect types of nagA, nagB, ldhA, msccg and msccg2 in a genome and two gene single base mutation types of odhA and glnA, and the codon of the glnA for coding the 405th amino acid is mutated from TAC to TTC. By blocking excretion of glutamic acid of corynebacterium glutamicum and enhancing conversion of glutamic acid to glutamine, the supply capacity of a precursor of corynebacterium glutamicum is enhanced, and an optimal way for synthesizing N-acetylglucosamine is obtained, so that efficient production of N-acetylglucosamine in corynebacterium glutamicum is realized, and the yield of N-acetylglucosamine in corynebacterium glutamicum is increased. The method has important economic value and social significance.

The invention discloses an emodin anthraquinone high-yield engineering strain. According to the invention, mdpG and mdpF genes of aspergillus nidulans are exogenous introduced into a chassis strain aspergillus nidulans LO8030, so that the genetically engineered bacterium for producing emodin is obtained. Furthermore, the g1234 gene is knocked out from the genetically engineered bacterium for producing the emodin, so that the yield of the emodin can be obviously improved. The decarboxylase gene is further introduced in an exogenous manner, so that the yield of the emodin can be further remarkably improved. And on the basis, exogenous introduction of mdpK, mdpC and mdpB genes is carried out, such that the chrysophanol-producing genetically engineered bacterium is obtained. Or the rugG gene from talaromyces sp.YE3016 is introduced in an exogenous manner, so that the genetically engineered bacterium for producing the muscardine hydrochloride is obtained.

The invention relates to an L-valine production strain as well as a construction method and application thereof, the strain is obtained by transformation through a metabolic synthesis path of L-valine, and the construction method comprises the following steps: by taking Cornebacterium glutamicum ATCC 13032 (C.glutamicum) as an original strain, carrying out editing transformation such as gene knockout, gene overexpression, gene heterologous expression, gene double-copy expression and the like on a genome of the original strain by utilizing a metabolic engineering means. The L-valine production strain disclosed by the invention can be used for synthesizing L-valine from the beginning by taking glucose as a carbon source, has the advantages of high production rate, short fermentation period, high strain stability and low cost, and lays a foundation for realizing large-scale production of L-valine. According to the fermentation tank fermentation method, the L-valine is produced in a mechanical stirring type fermentation tank by using the L-valine production strain Va1-09, 86.4 g / L of L-valine is produced by fermenting for 36 hours, and the method has a good industrial prospect.

The invention discloses an E3 ubiquitin enzyme mutant as well as modification and a protein targeted degradation system and application thereof, and particularly relates to a series of targeted protein degradation enhanced E3 ubiquitin enzymes (Trim21-N) and a modification method thereof, the E3 ubiquitin enzymes (Trim21-N) are obtained by modifying a wild type Trim21 (wtTrim21) protein structural domain, and the E3 ubiquitin enzymes comprise Trim21-a, Trim21-b, Trim21-c, Trim21-d, Trim21-e and Trim21-f. The efficiency of degrading the target protein through the ubiquitin proteasome is remarkably improved, and the method has the characteristics of strong endogenous and exogenous universality, high specificity, ubiquitin proteasome dependence, low cytotoxicity, strong cell compatibility and the like on the degradation of the target protein. The invention provides E3 ubiquitin enzyme (Trim21-N) based on a targeted protein degradation enhancement version and an exogenous fluorescence report degradation system of the E3 ubiquitin enzyme (Trim21-N), preferably Trim21-e with the optimal degradation effect, and the E3 ubiquitin enzyme (Trim21-N) is named as delta Trim21. The invention further provides an endogenous protein degradation system based on the delta Trim21. The method has the characteristics of high specificity, rapidness and small module, so that the method can be used for researching various biological and disease mechanisms, including the fields of cell targeted therapy, cell mechanism exploration, genetic engineering and the like.

The invention particularly relates to a saccharomyces cerevisiae genetically engineered bacterium for de novo synthesis of 7-DHC as well as a construction method and application of the saccharomyces cerevisiae genetically engineered bacterium and belongs to the technical field of biology. According to the invention, a saccharomyces cerevisiae engineering bacterium SC9 is taken as an original strain, multiple times of multiple copies of tHMG1, DHCR24 and ERG1 are carried out on TY transposon, multiple copies of ERG2, POS5, ERG7, ERG11, IDI1 and ERG3 are carried out on TY transposon, and POS5, ERG3 and DHCR24 are singly copied on a genome, so that the content of intracellular reducing coenzyme II is increased, and a main path for synthesizing 7-DHC is enhanced. The genome single copy ACL, ACS1, ERG10, ERG19 and the like are used for enhancing the utilization of saccharomyces cerevisiae ethanol and dynamically regulating and controlling the synthesis of 7-DHC. The saccharomyces cerevisiae gene engineering strain constructed by the invention has high product accumulation, and lays a foundation for replacing a traditional multi-step chemical synthesis route and realizing industrial production.

The present disclosure relates to isolated nucleic acid molecules comprising at least two double-stranded inhibitory ribonucleic acid (RNA) molecules adapted to silence the same or different genes by RNA interference to enhance silence, thereby modulating gene expression.

The invention relates to a method for increasing the yield of L-malic acid produced by fermentation by adding ethylene glycol, which comprises the following steps: constructing genetic engineering escherichia coli, and introducing ethylene glycol metabolism genes (overexpression) including glycerol dehydrogenase, lactaldehyde reductase, lactaldehyde dehydrogenase and the like; and overexpression of other genes involving malic acid production, such as malic acid synthase, phosphoenolpyruvate carboxylase and malic acid export protein. The gene engineering strain is an E.coli BW25113 strain of which ldhA, pta, poxB, adhE, pflB, mgsA, frdA, maeA, maeB, yqhD, ycdW and iclR genes are substantially inactivated, and the deleted genes are beneficial to reducing the generation of metabolic byproducts and increasing the accumulation of malic acid. The genetic engineering strain is subjected to fermentation culture by taking glucose and ethylene glycol as carbon sources, and L-malic acid is separated from a culture after culture is finished.

The present disclosure relates to compositions and methods of increasing levels of fetal hemoglobin (HbF) in cells. The present disclosure further relates to methods for treating patients suffering from blood cell diseases, including those associated with reduced amounts of functional adult hemoglobin (HbA), such as sickle cell disease and β-thalassemias

The present invention provides a genetically engineered bacterium for synthesizing N-acetylglucosamine, a preparation method thereof, and an application thereof. The genetically engineered bacterium uses Escherichia coli as an initial bacterium, and related genes for the catabolism of N-acetylglucosamine are knocked out on the genome of the initial bacterium, and related genes for the metabolic synthesis of N-acetylglucosamine are overexpressed, so as to improve the production level of N-acetylglucosamine. Among them, the related genes for the catabolism of N-acetylglucosamine include any one or a combination of at least two of nagB, nagA, manX, nagE, nagK, or ptsG, and the related genes for the metabolic synthesis of N-acetylglucosamine include any one or a combination of at least two of glmS, gna1, glnA, yqaB, icaC, galP, or glk.

The invention relates to construction of a Ralstonia strain capable of producing isobutanol by utilizing fructose and carbon dioxide and application of the Ralstonia strain. The construction method comprises the following steps: firstly, knocking out bkdA and bkdB genes in tricarboxylic acid cycle of the Ralstonia H16 to obtain a recombinant strain H1601; then, a PHB operator phaCAB in the H1601 is knocked out, and an alsS gene of bacillus subtilis and ilvC and ilvD genes of Ralstonia are introduced into the site, so that a recombinant strain H1602 is obtained. And finally, introducing yqhD and kivD gene expression cassettes of escherichia coli into the H1602, and constructing a recombinant strain H1603 capable of synthesizing isobutanol by using fructose and carbon dioxide. The strain can be used for synthesizing isobutanol through heterotrophic or autotrophic fermentation by taking carbon dioxide and fructose as carbon sources, the operation is simple and convenient, and higher yield can be obtained.

The invention belongs to the technical field of biology, and discloses a yarrowia lipolytica engineering strain for efficiently producing eicosapentaenoic acid based on citric acid metabolism regulation, a method and application of the yarrowia lipolytica engineering strain. Delta9 elongase, Delta8 desaturase, Delta5 desaturase and Delta17 desaturase genes in a fatty acid synthesis route, heterologous ATP-citrate lyase and a mitochondrial citric acid vector are overexpressed. According to the invention, ATP-citric acid lyase and a mitochondrial citric acid carrier gene which are respectively derived from Schizochylene sp. And Saccharomyces cerevisiae are jointly expressed in the yarrowia lipolytica engineering strain, so that the yields of EPA and grease in the yarrowia lipolytica engineering strain are remarkably improved, and compared with a control strain, the yields of the grease and the EPA are respectively improved by 243.35% and 161.85%.

The invention relates to recombinant saccharomyces cerevisiae for de novo synthesis of mevalonic acid as well as a construction method and application of the recombinant saccharomyces cerevisiae, and belongs to the technical field of genetically engineered bacteria. The technical problem to be solved by the invention is to provide the recombinant saccharomyces cerevisiae for de novo synthesis of mevalonic acid. According to the recombinant saccharomyces cerevisiae, saccharomyces cerevisiae is used as a chassis strain, and ACL, CTP1, RtME and MDH3 are over-expressed. A series of recombinant saccharomyces cerevisiae strains with high yield of mevalonic acid are constructed through a gene recombination method, the strains can be fermented in a conventional culture medium to prepare mevalonic acid, the fermentation process is simple, the cost is low, the yield is high, and the maximum yield of mevalonic acid can reach 5692.45 mg / L. According to the invention, the recombinant saccharomyces cerevisiae strain is used as a chassis strain, and recombinant genetically engineered bacteria for producing the santalene, the santalol and the patchouli alcohol are respectively obtained through gene recombination, so that the yields of the santalene, the santalol and the patchouli alcohol produced by fermentation are improved.

A method for treating a flavivirus infection includes administering to a subject in need thereof a composition which includes at least one of gentamicin sulfate, netilmicin, tobramycin, paromomycin, amikacin, capreomycin, trifluoperazine, dihydrostreptomycin, hydroxychloroquine, thioridazine HCl, efavirenz, miltefosine, nystatin, micafungin, bleomycin, resminostat, montelukast, norfloxacin, nedaplatin and cephalothin, which can exhibit excellent growth inhibition or killing effects of various species of flavivirus genus.