33results about How to "Improve fermentation yield" patented technology

The invention relates to the technical production field of metabolic engineering and genetic engineering, and particularly provides a uracil production strain as well as a construction method and application thereof. According to the strain, a uridine kinase gene udk is knocked out on an E.coli W3110 genome, and meanwhile, a T7RNA polymerase gene is integrated; the uridine monophosphate kinase gene pyrH is weakened; the method comprises the following steps: overexpressing a carbamyl phosphate synthetase gene carAB, a ribose phosphate pyrophosphate kinase gene prsA, a pyrimidine-5 '-nucleoside enzyme gene ppnN, UMP phosphohydrolase genes umpH and umpG, a ribonucleoside hydrolase 1 gene rihA and a glutamine synthetase gene glnA; an orotic acid nucleoside monophosphate decarboxylase gene pyrF and an orotic acid ribosyltransferase gene pyrE of wild type bacillus subtilis B.subtilis 168 are introduced in a heterologous manner, so that an orotate nucleoside monophosphate decarboxylase gene pyrF and an orotate ribosyltransferase gene pyrE are obtained. The safe, efficient and low-cost production of the uracil is realized.

The invention provides a method for increasing the fermentation yield of L-lysine, and belongs to the field of L-amino acid fermentation. The method for improving the fermentation yield of L-lysine comprises the following steps: preparing a shake-flask culture solution, preparing a seed solution, and carrying out fermentation culture. According to the method for improving the fermentation yield of the L-lysine, the tolerance of thalli to high osmotic pressure can be effectively improved, the acid production performance in the later period of fermentation is improved, and the single-tank yield and the sugar-acid conversion rate of the L-lysine are improved; meanwhile, the adverse effect of the osmotic pressure regulator on the fermentation stability of the L-lysine is further reduced, and the problems of inhibition of thallus metabolism, prolonging of the fermentation production period and reduction of the fermentation efficiency due to excessive addition of the osmotic pressure regulator are effectively avoided.

The invention relates to the technical field of genetic engineering, and provides a method for improving the yield of lincomycin by modifying a streptomyces lincomycin SLCG6736 gene, the SLCG6736 gene in the streptomyces lincomycin is deleted through a genetic engineering way to obtain a lincomycin high-yield engineering strain, and the obtained strain is used for fermentation production of lincomycin. Wherein the nucleotide sequence of the SLCG6736 gene is as shown in SEQ ID NO. 1. The invention further provides application of the method. The lincomycin biosynthesis negative regulator SLCG6736 has the advantages that the lincomycin biosynthesis negative regulator SLCG6736 is screened in the research, the SLCG6736 gene is knocked out through a genetic engineering way, a lincomycin high-yield strain can be obtained, and a technical support is provided for improving the lincomycin fermentation yield in industrial production.

The invention discloses an inositol oxygenase mutant and application of the inositol oxygenase mutant in preparation of glucaric acid. The protein provided by the invention is as follows: one or more amino acid residues in the 151, 253, 119, 295, 63, 92 and 71 sites of an amino acid sequence of wild inositol oxygenase are replaced to obtain the protein with the same function; the invention provides an enzyme activity enhanced inositol oxygenase mutant which is used for fermentation production of glucaric acid, and the yield of the glucaric acid is improved by 1.5 times.

The present invention provides a method for increasing the fermentation yield of L-lysine, and belongs to the field of L-amino acid fermentation. The method for increasing the fermentation yield of L-lysine is composed of the following steps: preparing a shake flask culture solution, preparing a seed solution, and fermenting and culturing. The method for increasing the fermentation yield of L-lysine of the present invention can effectively improve the tolerance of the bacterial body to high osmotic pressure, improve the acid production performance in the late fermentation period, and improve the single-tank yield and sugar-acid conversion rate of L-lysine; at the same time, further reduce the adverse effects of osmotic pressure regulators on the fermentation stability of L-lysine, and effectively avoid the problem of excessive addition of osmotic pressure regulators inhibiting bacterial metabolism, extending the fermentation production cycle, and reducing fermentation efficiency.

The invention provides a recombinant Aspergillus niger strain with high yield of trans-aconitic acid, and belongs to the technical field of gene engineering. The recombinant aspergillus niger strain can simultaneously express aconitase and aconitic acid isomerase. According to the genetic engineering aspergillus niger strain provided by the invention, the trans-aconitic acid fermentation yield is generally higher than that of an original strain, and the strain with the highest yield can reach 51g / L on a shake flask level, so that the genetic engineering aspergillus niger strain has very high application value.

The invention provides a genetically engineered bacterium for producing lactose-N-neotetraose as well as a construction method and application of the genetically engineered bacterium. The genetically engineered bacterium meets the following conditions: (1) exogenous expression of an MFS transport protein, beta-1, 3-N-acetylglucosamine transferase and beta-1, 4-galactosyl transferase is carried out; and (2) over-expression of a galactoside osmose enzyme and a UDP-galactose-4-epimerase is carried out. According to the genetically engineered bacterium disclosed by the invention, the MFS transporter, the beta-1, 3-N-acetylglucosamine transferase and the beta-1, 4-galactosyl transferase are exogenously introduced, and the expression quantity of galactosidase and UDP-galactose-4-epimerase in a genome is enhanced, so that the residual quantity of LNT II in an LNnT synthesis route can be effectively reduced, and the fermentation yield of the LNnT is increased.

The invention discloses a microbial fermentation real-time monitoring and molecular analysis method based on a single-cell Raman spectrum technology. The method comprises the following steps: firstly, preparing basic gold nanospheres which are microcosmically spherical and have the particle size distribution of 20-40 nm through a chloroauric acid oxidation reduction method, and characterizing the morphology of the basic gold nanospheres; secondly, the surface of the basic gold nanosphere is coated with a silicon dioxide thin layer through tetraethoxysilane hydrolysis and polymerization reaction, and the silicon dioxide functionalized gold nanosphere with the core-shell structure is prepared to serve as the microorganism living cell probe. The probe and different microbial cells are co-cultured, Raman fingerprint spectrums of the cells are sensitively captured, the single-cell Raman fingerprint spectrums are deeply analyzed by adopting a machine learning technology, and differential analysis (including strain difference, environment difference, metabolic capability difference and the like) is combined to determine the single-cell Raman fingerprint spectrums of the cells. Molecular indexes of specific expression of the microbial cells in different industrial production processes and different culture periods are screened out. The invention provides a universal tool for accurately realizing real-time monitoring and molecular analysis of microbial cells in different industrial processes, and has a wide industrial application prospect.

The invention discloses a genetic engineering strain of high-yield secondary metabolite Fumagillin as well as a construction method and application of the genetic engineering strain, and the genetic engineering strain is obtained by taking aspergillus fumigatus as a chassis bacterium through hosA gene knockout or hosA catalytic active site mutation. According to the method, a hosA gene knockout strain (delta hosA) and catalytic active site mutation strains (HosAD133A, HosAH175A and HosAD210A) are constructed by means of molecular biology, and the mutant strains are subjected to liquid or solid fermentation under the culture condition of 37 DEG C, so that the yield of Fumagillin can be remarkably increased. According to the method disclosed by the invention, the inhibition on a Fumagillin biosynthetic pathway is relieved and the metabolic flux is enhanced by utilizing the activity change of histone deacetylase caused by hosA defects, and an efficient and directional production method for high-yield Fumagillin is provided, so that the method has an important industrial application value.

The invention discloses a bacterial strain for generating coronatine based on a new mechanism, and an efficient breeding method and fermentation application of a coronatine high-yield bacterial strain, and relates to the technical field of bioengineering. The preservation number of the bacterial strain for generating coronatine by a new mechanism is CGMCC (China General Microbiological Culture Collection Center) NO.33248, and the bacterial strain is named as Pseudomonas syringae XZY; the method comprises the following steps: mutagenizing XZY, and performing efficient directional breeding to obtain a coronatine high-yield strain carrying an alpha-ABR phenotype, the preservation number of the coronatine high-yield strain is CGMCC (China General Microbiological Culture Collection Center) NO.30035, and the coronatine high-yield strain is named as Pseudomonas syringae XZY02. The coronatine fermentation yield of the strain is greatly improved compared with the fermentation yield before mutation, and in addition, the invention further provides a brand new method for obtaining the coronatine high-yield strain through directional breeding based on improvement of threonine downstream metabolic flow.

The present invention provides a 5-methyluridine producing strain and a construction method and application thereof, wherein the strain utilizes CRIPSR / Cas9 gene editing technology to produce E.coli On the UR12 genome, the artificial promoter P BBa_j23114 Replacement of uridylate kinase gene pyrH The natural promoter of umpH , umpG and no Gene, to ensure bacterial growth while enhancing the accumulation of 5-methyluridine; heterologous introduction and multiple copies of flavin-dependent thymidylate synthase gene thyX , overexpression of alkaline phosphatase gene phoA, Synergistically enhance the 5-methyluridine synthesis pathway; finally, overexpression of the serine hydroxymethyltransferase gene glyA, The supply of CH2-THF, the coenzyme of the key enzyme ThyX, is maintained, and the obtained strain has good genetic stability, high fermentation yield, and can stably produce 5-methyluridine.

This invention discloses a fermentation device for edible fungi that can improve fermentation yield. It includes a fermentation chamber, with a fixed column slidably connected to one side of the top of the chamber. A sampling cylinder is fixedly connected to the bottom of the fixed column. A limiting mechanism is provided at the top of one side of the fermentation chamber. Heating plates are installed on both sides inside the fermentation chamber, and a temperature sensor is installed on one side of the top of the fermentation chamber. This invention utilizes a drive motor installed at the top of the fermentation chamber. The drive motor drives a first gear to rotate, which in turn drives a second gear on one side to rotate. The first and second gears then drive stirring rods at the bottom to rotate, and the actuating rods on the two sets of stirring rods are staggered. This allows for rapid dispersing and mixing of the edible fungi material inside the fermentation chamber, accelerating the movement between edible fungi molecules and increasing the reaction rate of the edible fungi.

The invention relates to the technical field of wine brewing processes, in particular to a beer preparation method which comprises the following steps: collecting residual liquid in the distillation process when grape distilled wine is prepared, filtering the residual liquid to remove solid impurities to obtain filtrate, and dehydrating the filtrate until water equivalent to 10-12% of the mass of the filtrate is reduced, so as to obtain the beer. Pure grape distillation raffinate is obtained; wort containing yeast and hops required by beer preparation is prepared; adding pure grape distillation raffinate into the wort, mixing, and carrying out anaerobic fermentation; and after the anaerobic fermentation is completed, filtering and clarifying to obtain the beer. According to the beer preparation method disclosed by the invention, the residual liquid is utilized, so that the cyclic utilization of resources is realized, the fermentation efficiency is improved, the alcohol yield is increased, the flavor complexity and layering sense of the beer are improved, and the overall sensory quality of the beer is improved.

The utility model relates to the field of feed fermentation devices capable of being layered, and discloses a feed fermentation device with a layered structure, which comprises a shell, a fixing frame is fixedly arranged on the lower end surface of the shell close to the outer surface, and placing plates are fixedly arranged on two opposite sides of the fixing frame. A small heater and a small refrigerator are fixedly arranged on the upper end faces of the two containing plates correspondingly, a temperature panel is fixedly arranged on the portion, close to the middle, of the outer surface of the outer shell, and an inner shell is fixedly arranged in the outer shell. According to the utility model, when the device is used, the inner shell, the partition plate, the stabilizing block and the filter tank are matched for use, feed needing to be deflated during fermentation is placed in a space above the partition plate, and feed needing to be discharged with waste water during fermentation can be placed in a space below the partition plate; two different kinds of fermented feed can be put into one device for fermentation, the number of fermentation devices which can be placed in a plant is increased, and the fermentation yield of each batch of feed is increased.

The invention provides a strain for producing L-isoleucine and a construction method and application of the strain. Pck genes of the strain are knocked out, ilvBN (G20D, I21D and I22F), ilvA (V140M, F383A), lysC (T311I), hom (G378I), thrB (A20G), ppc, aspB, thrC, asd and brnFE are over-expressed, and the strain is prepared through operons Psod-ilvBN (G20D, I21D and I22F)-ilvA (V140M, F383A)-Ptuf-thrA-thrB-thrC and Psod-ppc-aspB-Ptuf

This invention belongs to the field of microbial polysaccharide fermentation and processing technology, specifically relating to a liquid sizing agent, its preparation method, and its application. This invention optimizes the fermentation process of *Sphingomonas sphingosine monophosphate* CGMCC No. 9531 to prepare a high-yield, high-viscosity fibrous sizing agent. Then, using vegetable oil and / or mineral oil as the dispersion medium, and in conjunction with a composite emulsifier, a high-concentration liquid sizing agent is prepared. This invention solves the problems of existing sizing agents being only solid powders with poor solubility and difficulty in maintaining high-concentration sizing agents stably in liquid systems. The high-concentration liquid sizing agent of this invention allows for pumping and precise control, meets product performance requirements, can be stably stored for more than 12 months, dissolves quickly, and remains uniformly stable after 72 hours of standing. The fermentation yield is increased by 25% compared to existing processes.

The invention belongs to the field of microbial fermentation, and particularly relates to a method for increasing the yield of L-threonine in a production strain and a recombinant strain. The invention provides a method for improving the yield of L-threonine in a production strain. The method comprises the step of knocking out a nucleotide sequence, which is not overlapped with an aspX gene and an aspA gene, in the L-threonine production strain. By knocking out the non-overlapping part of aspX and aspA, the transcription of aspX is stopped, and the yield of L-threonine is increased. The invention also provides an L-threonine high-yield recombinant strain and a method for producing L-threonine.

The application discloses a recombinant Yarrowia lipolytica engineering bacterium for synthesizing borneol from scratch, a construction method and application thereof. The recombinant Yarrowia lipolytica engineering bacterium is constructed by integrating a multi-gene tandem expression cassette containing an orange-flower base pyrophosphate synthase coding gene, a borneol base pyrophosphate synthase coding gene and a hydrolytic enzyme coding gene into a genome H1 site of Yarrowia lipolytica ATCC 201249 by CRISPR-Cas9 gene editing technology. The application first constructs the recombinant Yarrowia lipolytica engineering bacterium for synthesizing borneol from scratch by using a simple carbon source. The engineering bacterium does not need to add any terpene precursor externally, and only uses simple carbon sources such as glucose and sucrose as substrates. The borneol is synthesized from acetyl coenzyme A, a primary metabolic starting material, by a complete MVA pathway and an artificially constructed borneol-specific synthesis pathway, so that the whole biosynthesis of borneol is realized, and the limitation of the prior art on external precursors is broken through.

The invention relates to the technical field of fermentation devices, and particularly discloses a fermentation device capable of conveniently controlling a fermentation process and rapidly cooling, the fermentation device comprises a fermentation assembly, the outer wall of the fermentation assembly is provided with a driving assembly, the inner wall of the fermentation assembly is provided with a stirring assembly, and the fermentation assembly is used for containing a fermentation material. The driving assembly is used for driving the stirring assembly to rotate, and the stirring assembly is used for stirring the fermented materials. The multiple fixing sleeves can drive the multiple first auger blades to rotate with the conveying pipe as the center, at the moment, the two fixing discs rotate along the inner walls of the connecting holes in the two ends of the fermentation cylinder, the multiple second motors drive the fixing sleeves to rotate, the first auger blades are driven by the fixing sleeves to rotate, and the first auger blades are driven by the second motors to rotate; the first auger blade can stir the excrement, meanwhile, the excrement can be further stirred through the stirring plate, and the conveying cylinder drives the second auger blade to rotate to mix and stir the excrement in the middle of the fermentation cylinder.

The invention provides triticale coarse cereal vinegar and a processing method thereof, and relates to the technical field of food processing. The triticale coarse cereal vinegar comprises the following raw materials in parts by mass: 30-50 parts of triticale, 5-15 parts of peas, 5-15 parts of broom corn millet, 5-15 parts of sorghum, 5-15 parts of chenopodium quinoa willd, 10-30 parts of highland barley and 20-40 parts of medium-high temperature yeast for making hard liquor. According to the method, the problem of incomplete fermentation caused by coarse cereal husks is solved, the fermentation efficiency and the acetic acid yield are improved, the content of bioactive substances in the finished product vinegar is increased, and the oxidation resistance of the coarse cereal vinegar is improved. The finished vinegar product is rich in nutritional ingredients, fine and mellow in taste and simple in manufacturing process, the production cost is reduced, and meanwhile the stability and consistency of the product are guaranteed. According to the invention, the nutritional advantages of various coarse cereals such as triticale, peas, broom corn millet, sorghum, quinoa and highland barley are combined, so that the finished vinegar is rich in various nutritional ingredients such as protein, dietary fibers, vitamins and minerals, and has a relatively high health-care value.

This invention discloses a recombinant *Escherichia coli* strain, its preparation method and application, and a method for synthesizing oryzanol A, relating to the field of bioengineering technology. The recombinant *E. coli* strain uses *E. coli* BL21(DE3) as the chassis cell and undergoes the following modifications (a) and (b): (a) knocking out the pfkA gene; or, knocking out the pfkA gene and mutating the 5′ untranslated region of the gltA gene from the sequence shown in SEQ ID NO: 1 to the sequence shown in SEQ ID NO: 2; (b) inserting exogenous genes agmA, agmB, agmC, agmE, and agmF. This recombinant *E. coli* strain exhibits rapid growth, a short fermentation cycle (within 72 hours), can utilize inexpensive and readily available culture media, and significantly increases the fermentation yield of oryzanol A compared to existing *E. coli* systems.

The invention relates to the technical field of microbial fermentation, and particularly discloses high-whiteness fermented sodium propionate and a preparation method thereof.The preparation method of the high-whiteness fermented sodium propionate comprises the steps that a propionibacterium seed solution is inoculated into a fermentation culture medium for anaerobic fermentation, stirring and uniform mixing are conducted, liquid caustic soda is added in a fed-batch mode to maintain the fermentation pH to 6.5-7.0, and after fermentation culture, the high-whiteness fermented sodium propionate is obtained. Adding modified bentonite, heating and stirring to obtain sodium propionate fermentation liquor; sequentially filtering the sodium propionate fermentation liquor through a ceramic membrane and an ultrafiltration membrane, evaporating and concentrating, adding acid to adjust the pH value, and spray-drying to obtain the high-whiteness fermented sodium propionate, the obtained high-whiteness fermented sodium propionate product is high in whiteness, high in light transmittance, high in yield and excellent in comprehensive performance, meets the national standard requirements of food safety, and has a wide market prospect.

The invention discloses a method for increasing the fermentation yield of natamycin, and belongs to the technical field of natamycin production. The method comprises the following steps: (1) shake flask seed culture: under a sterile condition, coating streptomyces gilvosporeus spores in an agar culture medium, placing the agar culture medium in an incubator for culture, preparing the cultured streptomyces gilvosporeus into a spore suspension, sucking the spore suspension, adding the spore suspension into a triangular flask in the seed culture medium, and culturing to obtain a seed solution; (2) fermentation culture in a fermentation tank: inoculating the seed solution to a fermentation culture medium, carrying out stirring culture, feeding glucose, amino acid and a citric acid residue protein solution in the culture process, and then supplementing meat and bone meal to obtain fermentation liquor containing natamycin. The amino acid and the meat and bone meal are added in the fermentation culture process of the streptomyces gilvosporeus, so that the yield of natamycin produced by the streptomyces gilvosporeus is remarkably increased.

This invention provides a uracil-producing strain, its construction method, and its application. The strain utilizes CRIPSR / Cas9 gene editing technology to produce... E. coli On the UR14 genome, firstly by knocking out psuG Gene, black Gene, rutA Genes and upp Genes were introduced to block the breakdown of uracil; then, the uridine phosphorylase gene was overexpressed. udp and pyrimidine-5'-nucleotide nuclease gene ppnN This synergistically enhances the synthesis and accumulation of uracil; finally, overexpression of the phosphoribosyltransferase gene... pgm, Further enhancing the conversion of the byproduct 1-phosphate ribose into the precursor 5-phosphate ribose-1-pyrophosphate improves carbon utilization. The resulting strain exhibits good genetic stability and high fermentation yield, enabling stable production of uracil and demonstrating broad application prospects.

This invention provides a method for modifying Streptomyces lincosinate. SLCG_3615 A method to increase lincomycin production through gene therapy involves knocking out the TetR family transcriptional regulatory genes in Streptomyces lincomyces via genetic engineering. SLCG_3615 A high-yield engineered strain of lincomycin was obtained, and the obtained strain was used to produce lincomycin through fermentation; among which, SLCG_3615 The nucleotide sequence of the gene is shown in SEQ ID NO.1. This invention also provides... SLCG_3615 Applications of gene knockout modified *Streptomyces lincomycin*. In this study, the negative regulator of lincomycin biosynthesis, SLCG_3615, was screened and knocked out on the chromosome of *Streptomyces lincomycin* via genetic engineering. SLCG_3615 SLCG_3615 Gene copying can yield high-yield strains of lincomycin, providing technical support for increasing the fermentation yield of lincomycin in industrial production.

The application discloses a culture method for improving the fermentation yield of tacrolimus and belongs to the technical field of tacrolimus production. The culture method for improving the fermentation yield of tacrolimus comprises the following steps: inoculating Streptomyces tsukuba into a fermentation culture medium for fermentation culture to obtain a tacrolimus fermentation liquor; and performing separation and purification on the tacrolimus fermentation liquor to obtain tacrolimus; and adding macroporous adsorption resin into the fermentation culture medium according to the fermentation time during the fermentation process. The method for improving the fermentation yield of tacrolimus controls the carbon and nitrogen source concentration and pH in the fermentation process and supplements macroporous resin, improves the secondary metabolism efficiency, adsorbs tacrolimus in the fermentation liquor through the macroporous adsorption resin, reduces the negative feedback inhibition effect on the secondary metabolism of the production strain caused by high-concentration tacrolimus, and significantly improves the tacrolimus yield.

The invention discloses an expression cassette, a genetically engineered bacterium for producing ergothioneine and application of the genetically engineered bacterium. The expression cassette comprises coding genes of Ncegt1, egtEncr and egtDAfr, and the coding genes of the egtEncr and the egtDAfr are shown in the description. The amino acid sequences of Ncegt1, egtEncr and egtDAfr are respectively as shown in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5. The expression cassette has the advantage of being capable of improving the fermentation yield of the ergothioneine, and the genetically engineered bacterium for producing the ergothioneine can autonomously produce histidine which is one of substrates, has the advantages of being high in fermentation yield, high in production intensity and low in cost when being applied to production, and has industrial application prospects.

The invention belongs to the technical field of gene engineering, and particularly relates to a histidine methyltransferase mutant and application thereof.The catalytic activity of the mutant protein is remarkably improved compared with that of parent protein, and then production of ergothioneine catalyzed by histidine methyltransferase is promoted; and a novel synthetic route and engineering bacteria are provided for production of ergothioneine.

The present invention provides a method for increasing the yield of erythromycin by modifying the SACE_5312 gene of Saccharopolyspora erythromycin, wherein the SACE_5312 gene of the TetR family in Saccharopolyspora erythromycin is deleted by a genetic engineering method, and then an erythromycin high-yield engineering strain is obtained, and the erythromycin high-yield engineering strain is used to ferment and produce erythromycin; wherein the nucleotide sequence of the SACE_5312 gene is shown in SEQ ID NO.1. The present invention also provides an application of the above method. The present invention has the advantages that the yield can be greatly increased by fermenting and producing erythromycin with the obtained erythromycin high-yield engineering strain, and new technical support is provided for increasing the yield of erythromycin in industrial production.

The invention relates to recombinant corynebacterium glutamicum as well as a method for producing glutathione through fermentation and application thereof. The recombinant corynebacterium glutamicum takes corynebacterium glutamicum as an expression host; a cysteine desulfatase gene and a glutamyltranspeptidase gene are knocked out, and a gene of a glutathione bifunctional enzyme mutant is over-expressed; according to the glutathione bifunctional enzyme mutant, glutathione bifunctional enzyme with the amino acid sequence shown as SEQ ID NO.2 is used as a parent, histidine at the 11th site of the parent glutathione bifunctional enzyme is mutated into proline, and asparagine at the 239th site of the parent glutathione bifunctional enzyme is mutated into glutamic acid. According to the recombinant corynebacterium glutamicum CG05, fermentation parameters such as temperature, pH and supplementary material concentration of a 5L fermentation tank are optimized, the highest yield of glutathione in fermentation liquor of the recombinant corynebacterium glutamicum CG05 is 25.78 + / -0.25 g / L (83.97 + / -0.81 mM), the biomass OD600 is 59.8, the fermentation yield is increased by 2.1 times compared with that of a CG05 strain before optimization, the yield is increased by 5.7 times compared with that of an original strain CG01, and the conversion rate is 83.97%.