41 results about "Actinoplanes" patented technology

The invention discloses a method for preparing acarbose, which comprises the following steps: (1) actinoplane slants are inoculated in a slant culture medium for slant culture; (2) slants with full growth are selected and inoculated in a seed culture medium for seed culture; (3) fermentation cylinder culture is carried out, and glucose and maltose are added into a fermentation culture medium in afluid mode in a culture process. The method is characterized in that a nitrogen source is also added in the fluid mode in the step (3); and the content of amino nitrogen in the culture medium after adding the nitrogen source in the fluid mode is controlled to be smaller than or equal to 0.15 mg / ml except 0 mg / ml. The method overcomes the defect of difficult separation of components C which are impurities in the prior art, replenishes the nitrogen source in the process of fermentation, controls the content of the nitrogen source and can reduce the formation of the impurities C while greatly improving the yield of the acarbose.

The invention discloses an echinocandin biotransformation method using an actinomycete whole cell or fermentation broth as a catalyst. The invention also provides an echinocandin biotransformation method using cyclodextrin and its derivative or other cosolvents. A transformation system involved in the invention is characterized in that the system can effectively improve the solubility of echinocandin compounds in an aqueous solution, improve the utilization rate and the reaction rate of substrates, and improve the transformation rate of products. Actinoplanes involved in the invention and their mutant strains are in a nutritional medium containing an assimilable carbon and nitrogen source and can generate whole-cell deacylated enzymes under ventilation conditions, and the whole cell deacylated enzymes can be used for the echinocandin biotransformation.

The invention discloses a gene engineering bacterium for the high efficiency conversion of echinocandin B and a preparation method thereof. The echinocandin B is an engineering bacterium of an expression cassette integrated with echinocandin B deacetylase gene in a genome of Actinoplanes utahensis wild strain. The conversion of echinocandin B verifies that the conversion efficiency of the gene engineering bacterium is 1.8 times the conversion efficiency of the wide stain.

The invention provides a microbial agent. The microbial agent comprises the following components in parts by weight: 20 to 60 parts of hydrogen-oxidizing bacteria, 20 to 60 parts of nitrogen-fixing bacteria, 30 to 50 parts of actinomycetes and 30 to 60 parts of fungi, wherein the actinomycetes can be actinoplanes, streptomycetes or nocardia; and the fungi can be pythium or trichoderma. The microbial agent disclosed by the invention generates a certain synergistic effect through the combination of the hydrogen-oxidizing bacteria, the nitrogen-fixing bacteria, the actinomycetes and the fungi, and can improve the yield of wheat, promote the development of wheat root systems and improve the good earth holding capacity of roots for soil, so that main roots, side roots and fibrous roots are deeper and wider in coverage and are difficult to lodge and stronger in water absorption, eustipes close to the ground has better toughness and strength, the toughness and strength of the stems can be improved, and the stems are difficult to break. The microbial agent provided by the invention can shorten the growth and maturation time, enable crops to be matured in advance, reduce the risks of vigorous growth of seedlings and green remaining and late maturation of plants, and promote early fruiting and maturation of the plants to appear in the market.

The invention discloses a Rapamycin biosynthesis gene from antinoplanes, and a separation method and application thereof. The gene is a nucleotide sequence shown by SEQ ID NO:1.

The invention discloses a method for fermenting acarbose. The method comprises the following steps of: (1) taking Actinoplanes species and inoculating into actinomyces seed culture medium to prepare seed solution; and (2) taking the seed solution prepared in the step (1), inoculating into an actinomyce fermentation medium, and fermenting, thereby obtaining acarbose fermentation broth, wherein selenium with the concentration of 2-21mg / L is added in the fermentation medium. The fermentation method disclosed by the invention has the advantages of increasing the yield of the acarbose, remarkably reducing the content of an impurity A in the acarbose and shortening the fermentation period.

The invention discloses actinoplanes CGMCC NO.8430 of high-yielding sirolimus, belonging to the technical field of fermentation engineering. The bacterium is generated by mutating actinoplanes hn-016 of high-yielding sirolimus screened in a soil sample of Hainan Island as an original strain by plasmas at constant pressure and room temperature. Compared with conventional mutation methods, the mutant strain BCStr-096 disclosed by the invention is higher in hereditary stability and higher in production capacity of sirolimus, and the output is improved by 175% compared with the output 200mg / L of the original strain hn-016. The output is improved by about 37% in the highest fermentation level of existing sirolimus, so that the production cost is greatly lowered.

It is intended to provide novel compositions usable in improving the flavor of an alcoholic beverage made from grapes typified by wine. Namely, a composition usable in improving the flavor of an alcoholic beverage made from grapes which contains the culture of a strain belonging to a genus Aspergillus, Penicillium, Rhizopus, Rhizomucor, Talaromyces, Mortierella, Cryptococcus, Microbacterium, Corynebacterium or Actinoplanes and being capable of producing diglycosidase.

The invention relates to an acarbose engineering bacterium, a method for preparing the engineering bacterium and an application of the engineering bacterium. The acarbose engineering bacterium provided by the invention is prepared by inactivating treY gene of actinoplanes by introducing homologous recombination fragments by using a conjugational transfer method. The content of the generated acarbose C-component impurity of the acarbose engineering bacterium after the acarbose engineering bacterium is fermented is greatly reduced.

The invention discloses an echinocandin biotransformation method using an actinomycete whole cell or fermentation broth as a catalyst. The invention also provides an echinocandin biotransformation method using cyclodextrin and its derivative or other cosolvents. A transformation system involved in the invention is characterized in that the system can effectively improve the solubility of echinocandin compounds in an aqueous solution, improve the utilization rate and the reaction rate of substrates, and improve the transformation rate of products. Actinoplanes involved in the invention and their mutant strains are in a nutritional medium containing an assimilable carbon and nitrogen source and can generate whole-cell deacylated enzymes under ventilation conditions, and the whole cell deacylated enzymes can be used for the echinocandin biotransformation.

The invention discloses a preparing method of high unit acar wave sugar through regulating and controlling microorganism metabolism of flowing actinomycete Actinoplanes species, which comprises the following steps: adopting computer automatic speed changing supplementary material method; supplementing the material from 40-48 h of microbe ferment; supplementing at 3-4 L feed liquid per min; persisting to 84-92 h; adjusting the quantity of supplementary material to 1-2 L feed liquid per minute. This sugar possesses high ferment level and low cost, which does not need to modify original device.

The invention discloses a high-yield Fidaxomicin genetically engineered bacterium Actinoplanes deccanensis YP-2 and a construction method. A positive regulation gene biosynthesized by Fidaxomicin is guided into starting bacterium Actinoplanes deccanensis YP-1, the high-yield Fidaxomicin genetically engineered bacterium Actinoplanes deccanensis YP-2 is sieved and obtained, the yield is improved by400% to 500% compared with the starting bacterium and reaches up to 130 mg / L, and the Fidaxomicin genetically engineered bacterium has extremely high application prospects. The method is efficient, accurate and convenient in operation, and a new research means is provided for the construction of efficient biosynthetic pathway of an actinomycetes drug. The Fidaxomicin genetically engineered bacterium Actinoplanes deccanensis YP-2 can be applied in the preparation of drugs for curing diarrhea caused by the infection of clostridium difficile.

The invention provides a bio-organic fertilizer containing composite chitin. The bio-organic fertilizer is prepared from the following raw materials of potassium fulvate, peat, oleanolic acid, steamed bone meal, feather, composite chitosan, deprodone propionate, epoxy resin powder, astragalus smicus plants, satsuma stone, soybean dietary fiber, streptoporangium roseum actinoplanes and halomonas halophiles. The invention also provides a preparation method of the bio-organic fertilizer. The fertilizer prepared according to the invention has the advantages that the effective viable count is 7.5 to 7.7 hundred million cfu / g; the total nitrogen content is 3.6 to 3.8 weight percent; the total phosphor content is 2.3 to 2.5 weight percent; the total potassium content is 2.7 to 2.9 weight percent; the organic matter (metered by dry basis) content is 47.7 to 47.9 weight percent; the moisture content is 21.4 to 21.7 weight percent; the pH is 5.64 to 5.68. The prepared fertilizer has the effect of improving the soil of carbonate saline-alkali land.

The invention relates to a culture medium for producing teicoplanin by using actinoplanes and a culture method. The culture medium comprises a seed culture medium and a fermentation culture medium. Byusing a culture medium recipe and the culture method provided by the invention, the teicoplanin fermentation unit can be improved; the fermentation period can be shortened; the fermentation cost canbe reduced; the environment influence on raw and auxiliary material sources is reduced to the maximum degree; the sufficient supply of raw and auxiliary materials is ensured; and the stable and efficient production of the teicoplanin is realized.

The invention discloses a method for obtaining rapamycin fermentation liquor by utilizing actinoplanes, belonging to the technical field of fermentation engineering. The method comprises the following steps of culturing actinoplanes CGMCC No.8430 on a slant culture medium, inoculating the cultured slant strain onto a shake flask seed culture medium, then inoculating the cultured seed liquor into a seeding tank, and then inoculating the seed liquor into a fermentation culture medium to undergo fermentation culture, thus obtaining the rapamycin fermentation liquor. The method has the beneficial effects that rapamycin with relatively high concentration can be generated from actinoplanes by optimizing the culture medium and controlling the culture conditions; the yield of rapamycin can be about 750mg / L in fermentation experiments in a 30L fermentation tank, and the yield of rapamycin can be about 840mg / L in fermentation experiments in a 200L fermentation tank, so that rapamycin more easily meets the requirement of industrial production; moreover, high concentration is more beneficial for subsequent extraction operation.

The invention discloses a method for converting echinocandin B through actinoplanes utahensis. The method comprises the following steps: (1) culturing actinoplanes utahensis so as to obtain a bacterial liquid, centrifuging to collect bacteria, suspending the bacteria in a buffer saline solution, adding a sodium alginate solution, uniformly mixing, stably injecting into a CaCl2 solution, performingimmobilization, and putting the immobilized beads into a column for later use; (2) dissolving echinocandin B into an organic solvent, putting into the buffer saline solution till the concentration ofthe echinocandin B is 0.1-5g / L, further putting the echinocandin B into the column obtained in the step 1), and performing conversion. The method has the remarkable advantages of being simple in process and low in cost, and the prepared immobilized cells have the advantages of good stability, high conversion rates and multiple times of repeated use.

The invention provides a rapamycin structural analogue and a preparation method thereof. The rapamycin structural analogue is as shown in a formula I. The preparation method comprises the following steps of performing fermentation culture on actinoplanes containing polyketide synthase rapA mutant genes, and obtaining the rapamycin structural analogue from fermentation liquor, wherein the polyketide synthase rapA mutant genes are as shown in a sequence table SEQ ID No.1. Compared with a conventional chemical synthesis method, the preparation method of the rapamycin structural analogue provided by the invention is simpler to operate, lower in cost, and smaller in environmental pollution; and besides, the rapamycin structural analogue prepared by the preparation method has good anti-fungal effects.

An Actinoplanes spp. genetic manipulation system introduces a phi derivative plasmid and a pIJ101 derived replicating plasmid into Actinoplanes spp. In a mycelium conjugal transfer mode in order to realize gene expression and knockout. The Actinoplanes spp. strain genetic manipulation system is established and systemically optimized, and a genetic manipulation tool suitable for like strains is developed and is successfully applied to accurate genome editing to improve the acarbose output and lays a foundation for realization of rational design and reconstruction of the Actinoplanes spp. strain. The system is adopted to reinforce expression of acarbose biosynthesis gene cluster (acb cluster) in order to increase of the final fermentation output of acarbose by 35%.

The invention discloses a method for knocking out negative regulatory genes to improve the fermentation level of acarbose. By adopting the method disclosed by the invention, the negative regulatory protein genes ACPL_5445 and ACPL_3989 are knocked out from actinoplanes to obtain acarbose high-yield mutant strains. By knocking out the negative regulatory protein genes, the expression of acarbose biosynthetic genes can be promoted, and finally the acarbose yield is obviously increased. Compared with an original strain, the high-yield strains obtained by the invention have the advantages that thefermentation yields are improved by 13% and 8%, and the laboratory shake flask fermentation levels reach 3.62 g / L and 3.47 g / L.

The invention relates to an acarbose-producing engineered bacterial strain, and a preparation method and application thereof, belonging to the field of bioengineering. The acarbose-producing engineered bacterial strain is an Actinoplanes strain for acarbose production or derivative strains thereof. In the strain, one or two selected from the group consisting of the following genes are inactivated:(i) a gene M coding a polypeptide with a sequence as shown in SEQ ID No. 2 or with at least 80%, 90%, 95% or 99% sequence identity to the sequence as shown in the SEQ ID No. 2; and (ii) a gene N coding a polypeptide with a sequence as shown in SEQ ID No. 3 or with at least 80%, 90%, 95% or 99% sequence identity to the sequence as shown in the SEQ ID No. 3. When the engineered bacterial strain provided by the invention is used for acarbose production, the content of an impurity component A and / or an impurity component B in acarbose is significantly reduced, so the quality of the acarbose is guaranteed; and a subsequent purification step is not needed or a purification step is relatively simple, so process steps are decreased, and cost is reduced.

The invention discloses a novel antibacterial and antiviral marine actinomycete. The actinomycete is prepared through the following steps: implanting streptomycetes, small single cell bacteria, Rhodococcus rhodococcus, Nocard's bacillus and Actinoplanes in maifanite used as a carrier, pouring the maifanite into a mixer at 20-37 DEG C, mixing the maifanite for 30 min, and culturing the carrier attached with the above thalluses for 3-5 d. The antibacterial and antiviral principle of the marine actinomycete is generation of extracellular enzyme in the culturing of the streptomycetes, small singlecell bacteria, Rhodococcus rhodococcus, Nocard's bacillus and Actinoplanes attached to the maifanite. The marine actinomycete is a highly-adaptable decomposing bacterium, can resist a high temperature, a high pressure, a high salt and low nutrition, has no toxic or side effects, has anti-bacterium, antivirus, anti-Leptospira, detoxification, antitumor and leukemia cell-destruction effects, and can effectively improve the immunity.

The invention discloses a method for obtaining rapamycin fermentation liquor by utilizing actinoplanes, belonging to the technical field of fermentation engineering. The method comprises the following steps of culturing actinoplanes CGMCC No.8430 on a slant culture medium, inoculating the cultured slant strain onto a shake flask seed culture medium, then inoculating the cultured seed liquor into a seeding tank, and then inoculating the seed liquor into a fermentation culture medium to undergo fermentation culture, thus obtaining the rapamycin fermentation liquor. The method has the beneficial effects that rapamycin with relatively high concentration can be generated from actinoplanes by optimizing the culture medium and controlling the culture conditions; the yield of rapamycin can be about 750mg / L in fermentation experiments in a 30L fermentation tank, and the yield of rapamycin can be about 840mg / L in fermentation experiments in a 200L fermentation tank, so that rapamycin more easily meets the requirement of industrial production; moreover, high concentration is more beneficial for subsequent extraction operation.

The invention belongs to the technical field of biology, and particularly relates to a preparation method of 7-O-demethylated rapamycin. According to the method, actinoplanes are used as starting strains, and the biosynthesis of the increased 7-O-demethylated rapamycin is realized through shake flask fermentation or fermentation tank fermentation by selecting different types of cosolvents and different amino acids corresponding to different periods of addition. The preparation process of the 7-O-demethylated rapamycin is simple to operate, short in fermentation period and high in directionally synthesized purity and yield, and provides abundant raw materials for impurity research of the rapamycin.