71 results about "Saccharopolyspora" patented technology

Spinosyn biosynthetic genes from Saccharopolyspora spinosa, spinosyn producing microorganisms transformed with the biosynthetic genes, methods using the biosynthetic genes to increase production of spinosyn insecticidal macrolides, and methods using the genes or fragments thereof to change the products produced by spinosyn-producing microorganisms are provided.

The invention discloses gene engineering bacteria capable of highly producing pleocidin and a preparation method thereof. In the gene engineering bacteria, an expression box of NDP-glucose synthesized enzyme gene gtt of NDP-4-ketone-6-deoxidation-Dglucose and an expression box of genes of NDP-glucose reductase gene gdh are integrated in genomes of Saccharopolyspora spinosa wild bacterial strains. According to fermentation validation, the fermenting units of the pleocidin of the gene engineering bacteria is increased by more than three times compared with that of the wild bacterial strains.

The invention provides a method for producing pleocidin by adopting saccharopolyspora spinosa through fermentation. According to the method, saccharopolyspora spinosa Z68 is adopted as a fermentation strain, an improved fermentation medium is adopted, fermentation cultivation is carried out under the conditions that the temperature is 26-30 DEG C, the rotation speed is 150-220 rpm, the compression-ventilation ratio is 1 to (0.3-0.7), the pot pressure is 0.03-0.04 MPa, and the dissolved oxygen is controlled to be 40% or above, and pleocidin is extracted from the fermentation liquid. According to the method provided by the invention, staring from the physicochemical properties of the fermentation medium of saccharopolyspora spinosa, the fermentation medium with stable yield of pleocidin and little foaming amount, and a material supplementation method are provided. The soybean meal / soybean cake powder in the fermentation medium are replaced by high-temperature soybean cake powder, and the experiment proves that in the fermentation process, the high-temperature soybean cake powder has small possibility of foaming compared with the soybean meal and low-temperature soybean cake powder, while part of fat not available in the high-temperature soybean cake powder but available in the soybean meal and the low-temperature soybean cake powder can be added from the outside in the middle and later periods of fermentation, and the adding at the time can achieve the effect of eliminating foams and solve the problem that the carbon source is insufficient in the later period.

The invention discloses a process for extracting pleocidin from fermentation liquor of saccharopolyspora spinosa, which comprises the following steps of: (1) preprocessing fermentation liquor; (2) adding a high dielectric constant polar organic solvent into the fermentation liquor for soaking and extracting the pleocidin, performing solid-liquid separation and collecting clear leach liquor; (3) removing the high dielectric constant polar organic solvent by vacuum concentration and volatilization to obtain the concentrated solution of the pleocidin; (4) adding a low dielectric constant or highalcohol extraction solvent for extraction to obtain a loaded organic phase; (5) adding acid water into the loaded organic phase to perform reextraction, and collecting a reextraction phase; and (6) removing the residual extraction solvent in the reextraction phase by volatilization, adjusting the pH value to 8.5 to 11.5 by using solution of sodium hydroxide (NaOH), allowing the pleocidin to precipitate, filtering, washing the pleocidin precipitate for 1 to 3 times by using diluted alkali liquor, and drying the washed product under vacuum to obtain pleocidin powder. The process for extracting pleocidin from fermentation liquor of saccharopolyspora spinosa has a total yield of over 80 percent and has the advantages of high efficiency, high quality, low cost, easy industrialization and the like.

The invention provides, biologically active spinosyns, hybrid spinosyn polyketide synthases capable of functioning in Saccharopolyspora spinosa to produce the spinosyns, and methods of controlling insects using the spinosyns.

The invention relates to a Saccharopolyspora spinosa high-pleocidin-yield engineering strain and application thereof. The Saccharopolyspora spinosa high-pleocidin-yield engineering strain is Saccharopolyspora spinosa S.sp-DELTApepA which is collected by China Center for Type Culture Collection on June 8th, 2015, and the culture collection number is CCTCC NO:M2015362. The pleocidin yield of the Saccharopolyspora spinosa S.sp-DELTApepA is enhanced to 137% of the initial strain.

The invention discloses a method of increasing the yield of spinosad by improving the fermentation condition of saccharopolyspora spinosa based on metabonomics. The method comprises the following steps: (1) determining intracellular metabolite of the saccharopolyspora spinosa; (2) analyzing the capability of the saccharopolyspora spinosa of producing the spinosad; (3) carrying out PLS analysis; and (4) carrying out process analysis. According to the method disclosed by the invention, the metabolite change law in the process of producing the spinosad by virtue of fermentation of the saccharopolyspora spinosa is revealed by utilizing the metabonomics means and combining multivariate statistics, and a metabolic change mechanism related to production of the spinosad is further explained; by analyzing the spinosad production capability and the metabolic level of the saccharopolyspora spinosa in different growth environments, the metabolin related to the production of the spinosad is found, thus providing a direction for optimizing the culture process of the saccharopolyspora spinosa and increasing the fermentation yield of the spinosad. The invention also provides new thinking and method for study in other culture technological processes of producing macrolides microorganisms.

The invention discloses a genetically engineered bacterium capable of increasing the yield of spinosads as well as a construction method and an application. The construction method of the genetically engineered bacterium capable of increasing the yield of spinosads comprises the following steps: (1) inserting an spnk expression cassette into an escherichia coli-saccharopolyspora spinosa shuttle plasmid pOJ260 to construct a recombinant plasmid pLU101; (2) guiding the recombinant plasmid pLU101 into saccharopolyspora spinosa ATCC49460, and integrating the recombinant plasmid with a genome through homologous recombination, thus obtaining the genetically engineered bacterium capable of increasing the yield of spinosads and naming the genetically engineered bacterium LU101. The genetically engineered bacterium constructed by the method can increase the yield of spinosads.

The invention discloses saccharopolyspora for decreasing biogenic amine and application of the saccharopolyspora, and belongs to the technical field of food fermentation. Saccharopolyspora hirsute J2capable of decreasing the content of the biogenic amine is screened from wheat koji and is applied to a brewing process of liquors (white spirits and yellow rice wine), fermented sausages or soy sauce, so that the content of the biogenic amine can be decreased, meanwhile, the content of amino acids can be increased, the nutrition value of a fermented product can be increased, and therefore, the saccharopolyspora is capable of improving the quality and safety of fermented foods and has wide application prospects.

The invention relates to saccharopolyspora spinosa strain, application of the saccharopolyspora spinosa strain and a method for preparing spinosad. The strain related to the invention has the preservation number of CGMCC No.3460. The invention also relates to a mutant strain obtained by the strain. The invention also relates to the method for preparing the spinosad, which comprises the following steps of: a, fermenting the saccharopolyspora spinosa strain or the mutant strain thereof disclosed by the invention to obtain fermentation liquor containing the spinosad; and b, separating the spinosad from the fermentation liquor. The strain disclosed by the invention has strong capability of producing the spinosad and has high industrial application value.

The invention provides a two-stage dissolved oxygen control method for increasing pleocidin output. The method is characterized in that the volumetric mass transfer coefficient kLa of oxygen is adjusted and controlled by adjusting throughput and stirring speed at the beginning of fermentation of saccharopolyspora spinosa in the process of preparing saccharopolyspora spinosa fermentation liquor, thus the initial oxygen volumetric mass transfer coefficient kLa can be controlled to be 100-130h<-1> and the accumulation of biomass of saccharopolyspora spinosa is promoted; then the volumetric mass transfer coefficient kLa of oxygen is adjusted and controlled by adjusting throughput and stirring speed when the fermentation enters the final cell growth exponential phase, and thus the initial oxygen volumetric mass transfer coefficient kLa can be controlled to be 60-92h<-1> and the synthesis and accumulation of pleocidin are promoted. According to the method, the two-stage dissolved oxygen control is performed, which obviously increases the pleocidin output and shortened the fermentation cycle. The two-stage dissolved oxygen control and fermentation method is easily realized and has a high application value.

The invention provides a construction method of spinosad heterologous expression strain, a spinosad heterologous expression strain obtained by the method and use thereof in preparing spinosad. The method utilizes a plurality of homologous recombination to replace the erythromycin synthetic gene cluster of Saccharopolyspora erythraea with the spinosad synthetic gene cluster and the rhamnose synthetic gene cluster, such that the Saccharopolyspora erythraea produces spinosad.

The invention discloses saccharopolyspora spinosa for producing spinosad and belongs to the field of microorganisms. The strain number of the saccharopolyspora spinosa BCSsBFSg<5>-006 is CGMCC (China general microbiological culture collection center) No.5546, and a fermentation medium for the saccharopolyspora spinosa of the spinosad comprises glucose 40g / L, soluble starch 30g / L, corn steep liquor 15g / L, cottonseed concentration powder 20g / L, yeast powder 20g / L and calcium carbonate 1g / L, and is prepared by distilled water with the pH (potential of hydrogen) of 7.0-7.2 regulated by NaOH 1mol / L. Compared with conventional spinosad production strains, the mutant strain BCSsBFSg<5>-006 is higher in spinosad production capacity, and accordingly yield of the spinosad can be effectively increased, and production cost is reduced.

The invention relates to a culture medium and a culture method for producing spinosad through fermentation of saccharopolyspora spinosa. A carbon source, a nitrogen source and optimal compatibility ratio in a primary seed medium, a secondary seed medium and a fermentation medium for production of spinosad through fermentation of the saccharopolyspora spinosa in a three-stage fermentation mode areconfirmed. By utilizing the culture medium formula and the fermentation control process provided by the invention, the fermentation unit of rice spinosad can be improved, the fermentation cost is reduced, the sources of raw materials and auxiliary materials are furthest reduced and are not influenced by the environment, sufficient supply of the raw materials and the auxiliary materials is ensured,and stable and efficient production of the spinosad is realized.

The invention discloses a spinosad high-yield gene engineering bacterium. The preservation number of the gene engineering bacterium is CGMCC No. 14193. The invention further discloses a method by enhancing a fatty acid metabolism pathway to construct the spinosad high-yield gene engineering bacterium and application of the engineering bacterium to produce spinosad. According to the construction method, by conducting expression enhancement on the oxidation pathway of fatty acid Beta of saccharopolyspora spinosa, supply for the spinosad to biosynthesize the important precursor of acetyl coenzyme A is improved, and compared with original strains, the yield of the spinosad of obtained gene engineering strains is improved obviously.

The invention discloses a saccharopolyspora spinosa recombinant strain with double bldD genes. The provided saccharopolyspora spinosa recombinant strain with double bldD genes is prepared by integrating bldD gene into saccharopolyspora spinosa genome. The provided saccharopolyspora spinosa recombinant strain with double bldD genes is named as S.spinosa-BldD, and is preserved in the China Center for Type Culture Collection (CCTCC) in May 31st, 2015, and the preservation number is CCTCC No. M2015347. The PCR detection results show that bldD gene has been successfully integrated into a target bacterial strain chromosome. In the plate culture, people find that in BHI and TSB plates, the spores of recombinant strains are prominently inhibited. The results of shaking flask fermentation show that the spinosad yield of recombinant strain is increased by 1.35 times compared with that of a referential strain.

The present invention discloses saccharopolyspora jiangxiensis and applications thereof in food and belongs to the technical field of food fermentation. The saccharopolyspora jiangxiensis J3 is screened from wheat koji and deposited in China Center for Type Culture Collection on April 30, 2020, and has the deposit number of CCTCC NO:M 2020104. The strain is applied to a food fermentation system and results show that the strain does not affect normal fermentation of food, has effects of reducing content of biogenic amines in fermented alcoholic beverages, fermented food and fermented seasoningproducts, and has broad application prospects in the field of food brewing.

The invention provides a novel integrase and application thereof to genetically modifying saccharopolyspora spinosa. The integrase particularly comes from saccharopolyspora endophytica CCTCC AA208003, and exogenous DNA (deoxyribonucleic acid) can be specifically locally integrated on saccharopolyspora endophytica chromosome by the integrase via specific cis-form components. In addition, independent free replication regions of different strain sources are discovered in the saccharopolyspora spinosa by an inventor via research on genetic factors of different strains of the saccharopolyspora spinosa. The novel integrase and the application have the advantages that vectors constructed on the basis of the integrase and sequences of the replication regions can be used for genetically modifying the saccharopolyspora spinosa, accordingly, novel spinosad analogues can be possibly discovered, and the spinosad fermentation yields can be possibly increased; different resistance molecular markers can be carried by plasmids, accordingly, the novel integrase can be used for inter-species or intergenetic genome shuffling operation on the saccharopolyspora spinosa, and efficient genome shuffling can be implemented.

The invention relates to a method for improving a saccharopolyspora spinosa spinosad fermentation yield. The method utilizes mutagenesis treatment to screen a saccharopolyspora spinosa spinosad high-yield mutant strain with a good starch utilization capability. The mutagenesis treatment comprises at least two of UV-streptomycin combined mutagenesis, 5-fluorouracil mutagenesis, microwave-streptomycin combined mutagenesis and nitrosoguanidine-streptomycin combined mutagenesis. Preferably, the mutagenesis treatment comprises the above four mutagenesis methods. Preferably, the UV-streptomycin combined mutagenesis, 5-fluorouracil mutagenesis, microwave-streptomycin combined mutagenesis and nitrosoguanidine-streptomycin combined mutagenesis processes are orderly carried out. The broth of the obtained mutant strain has spinosad titer substantially higher than that of an original strain screened through mutagenesis and thus the method can improve a saccharopolyspora spinosa spinosad fermentation yield.

The invention relates to a fermentation medium for improving an erythromycin yield and a method using the same, belongs to the field of microbes and provides a Saccharopolyspora erythraea fermentation medium and a method for preparing erythromycin through the medium. Through use of a proper amount of amylase in the fermentation medium, optimization of components of a bacterial strain medium and a fermentation medium and control of other culture conditions, an erythromycin yield is improved. The fermentation medium has an erythromycin yield of 11200 micrograms per milliliter, has a simple formula, has a low cost and is suitable for industrial large-scale production.

The invention relates to a method for extracting spinosad from a Saccharopolyspora spinosa fermentation solution. The method comprises the following steps: pretreatment of the fermentation solution, active carbon column treatment for removing a lot of pigments and gummy impurities, extraction of spinosad, back-extraction, fine filtration, crystallization, separation and drying. A compound enzyme is adopted during the pretreatment to treat mycelia of the Saccharopolyspora spinosa, so the spinosad generated in the mycelia is released into the fermentation solution; and an active carbon column is used to remove a lot of the pigments, colloids and other impurities from the fermentation solution, so the improvement of the purity of the spinosad product is greatly benefited. The method has the advantages of simplicity in operation, short period, safety in process, low cost, high yield and high purity.

The invention relates to a method for extracting pleocidin from saccharopolyspora spinosa fermentation liquor. The method comprises the following steps: (1), pre-treating saccharopolyspora spinosa fermentation liquor and carrying out solid-liquid separation; (2), adsorbing centrifuged organic substances in a water phase by an adsorbent; (3), combining solids which are centrifuged and separated for the first time and the second time, and drying in vacuum; (4), extracting pleocidin from solids; and (5), recrystallizing by using a solvent to obtain a pleocidin product, decompressing and desolventizing. According to the method disclosed by the invention, a polar solvent is not needed to be added for extracting during pre-treatment, so that usage amount of an organic solvent is greatly reduced, pollution is reduced, cost is lowered, a centrifugal method can be adopted, operation is safe and separating effect is good; moreover, the adsorbent is added into the water phase for adsorbing organic substances in water, so that COD (chemical oxygen demand) in wastewater is greatly lowered, wastewater treating difficulty is reduced, the yield of the obtained product pleocidin is higher, and a total yield is 70%-95%; after the product is purified, the pleocidin content with mass content of 75%-93.2% is higher.

The invention discloses an insecticide for sugarcane planting and a preparation method of the insecticide. The insecticide for sugarcane planting is formed by fermenting compound bacteria and a Chinese herbal culture medium, wherein the compound bacteria are selected from one or two of saccharopolyspora spinosa and aspergillus niger; the Chinese herbal culture medium is prepared from, by weight, 20-40 parts of fructus forsythiae, 5-15 parts of garlic, 3-12 parts of celastrus angulatus rhizomes, 1-8 parts of fructus crotonis, 10-30 parts of stems and leaves of chrysanthemum coronarium, 30-50 parts of nutrients and 30-50 parts of deionized water. The insecticide for sugarcane planting and the preparation method of the insecticide have the advantages that effective components in Chinese herbal medicine are extracted through melanomyces and the saccharopolyspora spinosa, and secondary metabolic components generated in the fermentation process of the melanomyces and saccharopolyspora spinosa synergize with the active components contained in the Chinese herbal medicine so that the insecticide can reduce diseases and insect pests of crops to the maximum extent.

This disclosure concerns methods for producing a spinosyn producing strain that comprises modifying a nucleic acid molecule encoding for spinactin by introducing, mutating, deleting, replacing or inactivating a nucleic acid sequence encoding one or more activities encoded by said nucleic acid molecule. Methods for producing a modified Saccharopolyspora spinosa organism are also disclosed.

The invention provides, biologically active spinosyns, hybrid spinosyn polyketide synthases capable of functioning in Saccharopolyspora spinosa to produce the spinosyns, and methods of controlling insects using the spinosyns.

The invention discloses a Saccharopolyspora composition and application thereof in foods, and belongs to the technical field of food fermentation. Saccharopolyspora hirsute J2 and Saccharopolysporajiangxiensis J3 which are capable of decreasing the content of biogenic amine are screened from wheat koji and are prepared into a mixed Saccharopolyspora preparation, and the mixed Saccharopolyspora preparation is applied to a preparation process of fermented alcohol beverages, fermented foods or fermented seasonings, so that the content of the biogenic amine can be decreased, meanwhile, the contentof amino acids is increased, the nutritional value of a fermented product is increased, the quality and safety of the fermented foods are improved, and the application prospects is wide.

The invention relates to the technical field of microorganisms, and particularly discloses saccharopolyspora spinosa DS190375 as well as a fermentation product, a microbial inoculum, a fermentation and screening method and an application of saccharopolyspora spinosa DS190375. The preservation number of the saccharopolyspora spinosa DS190375 disclosed by the invention is CGMCC No. 18681. The fermentation method comprises the following steps: rejuvenating and screening the strain in a high-salt plate culture medium, and fermenting. The invention also provides a screening method of the saccharopolyspora spinosa with high spinosad yield, which comprises the following steps: screening to obtain the saccharopolyspora spinosa with high salt tolerance, and screening the obtained saccharopolysporaspinosa with high salt tolerance to obtain the strain with high spinosad yield. The saccharopolyspora spinosa DS190375 disclosed by the invention is resistant to high salt and high in spinosad yield.The fermentation method avoids the problem of unstable spinosad yield caused by strain performance degradation.

The invention provides saccharopolyspora spinosa capable of highly yielding spinosad and a method for increasing the spinosad yield of the strain. The saccharopolyspora spinosa capable of highly yielding the spinosad includes a J1-019-A strain and a spinosad producing strain saccharopolyspora spinosa capable of overexpressing spnJ or spnP; according to the J1-019-A strain, a polyketide synthase gene cluster which is not related to spinosad synthesis is knocked out from a genome of an original spinosad producing strain saccharopolyspora spinosa, and an attB site which can be conveniently transformed subsequently is introduced; and the spinosad producing strain saccharopolyspora spinosa capable of overexpressing spnJ or spnP performs overexpression on the spnJ or spnP in the original spinosad producing strain saccharopolyspora spinosa. The spinosad producing strain provided by the invention can realize fermentation production of spinosad through amplified fermentation of a fermentation tank, and can achieve the potential of industrial production.