37 results about "Arthrobacter simplex" patented technology

A biological dehydrogenation preparation method of steroid drug intermediate takes the formula (1) compound as a substrate, and adopts an arthrobacter simplex biological dehydrogenation method to obtain formula (2) compound; the special process comprises the following steps: smashing or dissolving the formula (1) compound with solvent, putting into a fermentation tank in which arthrobacter is cultivated for biotransformation, extracting, separating and refining, and drying to obtain dehydrogenated polymer, i.e. compound (2); and the dehydrogenation transformation rate can reach 70 to 90 percent.

The invention relates to a production method of steroids medicines, and concretely relates to a method for conversing 4-AD to produce 11 alpha-OH-ADD by mixed fermentation of two microbes. According to the invention, 4-AD is taken as a raw material, and the mixed fermentation is carried out through aspergillus ochraceus ATCC18500 and Arthrobacter simplex ATCC6946. Compared with hydroxylation and dehydrogenation reaction by individually using the two above bacteria, 11 alpha-OH-ADD can be obtained through mixed fermentation of 4-AD, two above bacteria have cooperative effect on the fermentation and conversion, The method has the following advantages: the overall yield is high, the conversion is more complete, a step of post-treatment is reduced, and the energy consumption is reduced. The mixed fermentation is capable of greatly reducing the production cost, increasing the conversion rate, reducing the conversion time, and relatively reducing the post-treatment and the energy consumption.

The invention discloses a salt-resistant ammonia-nitrogen-removing compound bacterial agent for printing and dyeing wastewater treatment. The salt-resistant ammonia-nitrogen-removing compound bacterial agent is prepared from, by mass, 5-8% of a saline-alkaline-resistant aliidiomarina haloalkalitolerans culture, 5-7% of a salt-resistant nesterenkonia halotolerans culture, 20-22% of a zobellella taiwanensis culture, 15-18% of a comamonas aquatica culture, 10-12% of a pseudomonas putida culture, 10-15% of a bacillus subtilis culture, 5-7% of an arthrobacter simplex culture and 17-18% of an enterobacter cloacae culture. The salt-resistant ammonia-nitrogen-removing compound bacterial agent for the printing and dyeing wastewater treatment contains various strains, is reasonable in compatibility,has a strong synergistic effect, can effectively remove ammonia nitrogen and industrial COD in printing and dyeing wastewater, has low input cost, and can be promoted and used on a large scale.

The invention discloses a method for preparing prednisone acetate. The method comprises the steps that firstly, positions 1,2 of cortisone acetate is dehydrogenated by using arthrobacter simplex CPCC140451 and are converted to generate the prednisone acetate, and then mixed solvents are used for refining. The method for preparing the prednisone acetate has the advantages of simple process, high refining efficiency, simple and easy obtained solvent, recycling and low toxicity, and can be used in large-scale industrial production.

The invention belongs to the technical field of bio-pharmaceuticals, and in particular relates to a method for producing a dexamethasone epoxide hydrolysate through one-step microbial fermentation and transformation. According to the method, with 16a-methyl-17[alpha],21-dihydroxypregna-4-ene-9,11-epoxy-3,20-dione-21 acetate as a substrate, the dexamethasone epoxide hydrolysate is directly produced mainly by conducting mixed fermentation by virtue of arthrobacter simplex and bacillus megatherium. In comparison with the prior art, the method provided by the invention, on the basis of a synergistic effect between mixed strains, integrally combines two steps of reactions, namely dehydrogenation and hydrolysis, so that the finished dexamethasone epoxide hydrolysate is directly obtained; and the product has the advantages of being short in production cycle, high in bio-transformation rate, high in yield, environment-friendly and low in cost.

The invention discloses a preparation process of a compound shown in a formula II. The compound is prepared from a compound shown in a formula I by biological fermentation and dehydrogenation, and in the processes of biological fermentation and dehydrogenation, an inert grinding material is added in fermentation liquid. In the invention, the fermentation manner is improved under a condition of using a common arthrobacter simplex strain; and a proper amount of inert grinding material is added in the fermentation liquid so that the yield and the quality of a compound product (namely a betamethasone intermediate) in the formula II are improved under the condition that the feeding concentration is maintained to be over 0.5%, and the production cost of betamethasone is reduced. In addition, no chemical solvent is used in the preparation process, and thus the cost for auxiliary materials is saved.

The invention relates to a construction method and application of an Arthrobacter simplex engineering strain high in organic solvent tolerance. The Arthrobacter simplex engineering strain is obtained by guiding a gene segment into Arthrobacter simplex; experiments verify that tolerance of the engineering strain obtained by utilizing the construction method to methanol and alcohol is improved obviously. By utilizing the engineering strain for steroid C1, 2 dehydrogenation reaction, adding amount of an organic solvent in a system is doubled, feeding concentration of a substrate is increased by three times, and generation amounts of products are increased in different degrees. By constructing and applying the converted engineering strain high in organic solvent tolerance, the adding amount of the organic solvent in the conversion system can be increased, the feeding concentration of the substrate is increased, the generation amount of the products is increased, and the engineering strain has important application value in increasing the conversion efficiency of high-concentration substrates of steroid hydrophobic compounds.

The invention belongs to the field of genetic breeding, and particularly relates to a mutant strain of arthrobacter simplex with stress tolerence and a genetically engineered bacterium constructed bythe mutant strain. The survival rate of the mutant strain under the impact conditions of 16% ethyl alcohol and 20% methyl alcohol is increased by 3.74 times and 2.10 times respectively compared with the original strain; the endurance capacity of engineering bacteria, overexpressing the genes of groEl, dnaK, recA, uvrD, katG, sod or treS from the mutant strain, to high-concentration organic solvents is obviously improved, and the endurance capacity to salt pressure and oxidative pressure is expresses to be higher. When the engineering bacteria are applied to steride C1,2 dehydrogenation reaction, the ethanol concentration in a system can be increased to 8 to 10%, the substrate CA concentration can be increased to 6 to 8g/L, the product yield can be improved by 0.41 to 3.56 times, and a goodbasis is established for the subsequent tolerance mechanism research and bacterial strain molecular modification and application.

The invention provides a production method of a methylprednisolone dehydrogenate. The production method of the methylprednisolone dehydrogenate comprises the following steps: A) adding water into a methylprednisolone Grignard substance, and pulping to obtain slurry; and B) biologically converting the methylprednisolone Grignard substance in the slurry into a methylprednisolone dehydrogenized substance by using arthrobacter simplex. According to the production method, no organic solvent is used, and the toxic effect on microorganisms is small during biotransformation; according to the method, the methylprednisolone Grignard substance is added with water and pulped into slurry, so that the feeding concentration and the substrate conversion rate of the methylprednisolone Grignard substance are greatly improved, the yield of a methylprednisolone dehydrogenized substance product is high, and the method is particularly beneficial to industrial production of the methylprednisolone dehydrogenized substance.

The invention relates to a production method of steroidal drugs, in particular to a method of preparing a boldenone steroidal compound by sequential fermentation of Arthrobacter simplex and a genetically-engineered yeast strain. The method comprises: providing a Pichia pastoris genetically-engineered strain capable of efficiently expressing 17beta carbonyl reductase; providing a process of fermenting the compound AD by microbial sequential conversion to prepare a compound BD, wherein the method refers to fermenting the AD via Arthrobacter simplex, deactivating the reaction system, and preparing the compound boldenone through sequential fermentation of the Pichia pastoris genetically-engineered strain.

The invention discloses a preparation method of prednisone acetate, and the method comprises the following steps: adding a hydrophilic organic solvent into Arthrobacter simplex fermentation broth, and adding 8-12% of prednisone acetate micropowder; finally, adding a lipophilic organic solvent for fermentation reaction; and after the fermentation reaction is finished, filtering or centrifuging the product, performing separating, decolorizing and crystallizing to obtain prednisone acetate. The quality of the product prepared by the preparation method is equivalent to that of the product in the prior art, but the yield is slightly higher, the single-batch yield is higher, the production efficiency is higher, and the preparation method is more suitable for large-scale industrial production.

The invention relates to a method for preparing 11alpha, 17alpha-dyhydroxyl-pregn-1,4-diene-3,20-dione by taking 17alpha-hydroxyprogesterone as a starting material and through mixed one-step fermentation adopting aspergillus ochraceus and arthrobacter simplex.

The invention relates to the technical field of pharmacy, in particular to a preparation method of a triketone dehydrogenation product, which comprises the following steps: mixing, stirring and pulping a triketone product and water to obtain slurry; adding a dispersing agent into the slurry, and sterilizing at high temperature to prepare triketone slurry; wherein the mass ratio of the triketone substance to the water is 1: (2-3); the mass ratio of the slurry to the dispersing agent is 1: (0.01-0.05); putting the triketone slurry into a fermentation culture solution of arthrobacter simplex for biotransformation in batches to obtain a triketone dehydrogenation product; wherein the total adding concentration of the triketone slurry is 2-4% of that of the fermentation culture solution. The biotransformation method can specifically remove 1, 2 hydrogen of 17 alpha-hydroxypregna-4-ene-3, 11, 20-triketone and introduce double bonds, the transformation rate is as high as 90%, the use of strong acid, strong alkali and toxic and harmful chemicals is avoided or reduced, the operation conditions are improved, the production cost is reduced, and the concept of green development is met.

The invention relates to an arthrobacter simplex engineered strain with excellent stress tolerance and a construction method and application thereof. The arthrobacter simplex engineered strain is obtained by adopting a promoter engineering method, a global transcription mechanism engineering method or a method combining the promoter engineering and the global transcription mechanism engineering. The strain still keeps good activity in a transformation system with high concentrations of organic solvents (8%) and substrates (15-45 g/L), with the product yield in a C1,2 (subscript) dehydrogenation reaction being 1.1-2.2 times higher than a control strain I (containing a wild-type promoter and a wild-type irrE). According to the construction method and the application of the engineered strainwith excellent stress tolerance, the concentrations of organic solvents and substrates in the transformation system are increased, and then the product production is increased. The invention has important guiding significance for improving the transformation efficiency of the high-concentration substrates of steroid hydrophobic compounds.

The invention relates to a genetically engineered bacterium and a leavening agent for dehydrogenation of steroid A ring 1,2 positions. The host bacterium of the genetically engineered bacterium is Arthrobacter simplex, the preservation number of which is ACCC 12070. The genetically engineered bacterium contains a recombinant gene fragment as shown in Sequence 2. The fermentation substrate of the genetically engineered bacterium contains cortisone acetate, fluoride cortical hormone and 4-AD. The constructed engineered bacterium provided by the invention performs fermentation by constant temperature ventilation stirring on the basis of yeast extract, phosphate, corn steep liquor and glucose containing nutritional ingredients. When the concentration of steroids is not higher than 10%(m/V), one time addition of steroid substrate is adopted, and?biological dehydrogenation conversion rate can reach to more than 95% in?32-48 hours. When the concentration of steroids is higher than 10%, continuous addition of steroid substrate is adopted, and biological dehydrogenation conversion rate can reach to more than 95% within 60 hours.

The invention belongs to the technical field of preparation of steroid hormone drugs, and particularly relates to a method for producing androstane dienedione by fermentation of arthrobacter simplex, which mainly comprises the following steps: optimizing fermentation culture medium components, feeding points and substrates for whole-cell transformation of arthrobacter simplex to obtain an ADD crude product with high conversion rate, and extracting and refining by mixing acetone and water to obtain the androstane dienedione. And an ADD fine product is obtained. The method effectively solves the problems that in the prior art, the conversion efficiency is not high, byproducts are difficult to separate, the process is tedious and not suitable for industrial mass production, and the like, the fermentation feeding concentration is increased to 30-60 g/L, and the conversion rate is increased to 97% or above.