31 results about "Actinoplanes sp." patented technology

This invention provides a process for producing a macrocyclic lactone compound, which comprises cultivating Actinoplanes sp. FERM BP-3832, in the presence of L-proline, L-hydroxyproline or L-nipecotic acid, and then isolating a macrocylic lactone compound from the fermentation broth. The compounds produced by this process include a compound of the following formula:The present invention also relates to a pharmaceutical composition comprising the same, which is useful as immunosuppressive, antimycotic, antitumor agent or the like.

A novel enzyme which has an activity to release side chain carboxyl groups and ammonia from a protein by acting upon side chain amido groups in the protein. This invention relates to a method for the production of an enzyme, which comprises culturing in a medium a strain that belongs to a bacterium classified into Cytophagales or Actinomycetes and has the ability to produce an enzyme having a property to deamidate amido groups in protein, thereby effecting production of said enzyme, and subsequently collecting said enzyme from the culture mixture. It also relates to a method for the modification of protein making use of a novel enzyme which directly acts upon amido groups in protein as well as to an enzyme which has a property to deamidate amido groups in protein and a gene which encodes said enzyme.

The invention discloses an alpha-amylase inhibitor producing strain, i.e. Actinoplanes sp.ZJB-08196 preserved in the China Center for Type Culture Collection located in Wuhan university, Wuhan Province, China on Feb. 16th, 2009 with the preservation number of CCTCC No. M209022. The alpha-amylase inhibitor is acarbose. The invention also discloses a screening method of the alpha-amylase inhibitor producing strain. The screening method provided by the invention is simple and effective, can screen a great amount of samples in a short time, has high sensitivity, reduces the long processes for processing and analyzing samples through HPLC (High-Performance Liquid Chromatography), and the like and improves detection efficiency.

The invention relates to a construction method for streptomycete expression plasmids and a production method for keratinase. The construction method for streptomycete expression plasmids includes the following steps: a promoter Xi of actinoplanes missouriensis xylose isomerase and a terminator of streptomycete avermitilis amylase are sleeved, and the promoter-shine-dalgarno (SD) sequence comes from 90 to 269bp of an actinoplanes missouriensis xylose isomerase gene; a cloned src family kinases (sfks) gene is inserted in a construction expression frame structure of a synthetic Xi promoter-SD-amyA2 terminator fragment, and then a conventional method is used for constructing the streptomycete expression plasmids. The production method for keratinase includes: leading the streptomycete expression plasmids into an expression host of streptomycete lividans TK24 through conjugal transfer, performing recombinant expression and generating the keratinase. Specific activity of a crude enzyme solution is 1700 U/mg after expression of the expression plasmids in the streptomycete lividans TK24 and is improved by 50 times compared with specific activity of starting strain streptomycete fradiae varieties S-221, and yield of the keratinase is much higher than starting strains after the expression.

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 relates to a preparation method of an actinomycetes fibrinolytic enzyme. Specifically, the method comprises the following steps: (1) adding ammonium sulfate powder to a fermentation supernatant containing actinomycetes fibrinolytic enzyme until a saturation degree is 25-35%W/V, standing by for a whole night at 4 DEG C and centrifuging so as to obtain a supernatant, adding ammonium sulfate powder to the supernatant once again until a saturation degree is 55-65%W/V, standing by for a whole night at 4 DEG C, centrifuging so as to obtain a precipitate, and dissolving the precipitate by virtue of a PB buffer solution so as to obtain crude enzyme liquid; and (2) centrifuging the crude enzyme liquid obtained from the step (1) at low temperature so as to remove impurities; purifying an obtained supernatant subsequently through Octyl-Sepharose FF hydrophobic interaction chromatography and Phenyl-Sepharose HP hydrophobic interaction chromatography so as to obtain a chromatographically pure enzyme; dialyzing and desalting the enzyme, and freeze-drying the enzyme so as to obtain fibrinolytic enzyme freeze-dried powder which can be preserved for a long time at minus 20 DEG C. The separation and purification method of the actinomycetes fibrinolytic enzyme disclosed by the invention has the advantages of simple operation steps and high recovery rate.

The invention provides a culture method for promoting an actinoplanes teichomyceticus industrial production strain to generate rich zoosporangiophore, in the formula of a strain culture medium, wheatgerm powder is selected as a main carbon and nitrogen source for spore germination, asparagine and ammonium sulfate are used as supplementary nitrogen sources, and trace elements supplement iron, manganese, cobalt and zinc ions. The culture temperature is controlled by stages. Glycerol and scleroglucan are used as composite cryoprotectants for -80 DEG C or ultralow-temperature liquid nitrogen preservation to prepare a strain library, so that the preservation stability of the strain library is greatly improved.

The invention discloses a method of utilizing an Actinoplanes sp. fermentation culture material to prepare three ramoplanin single components. The method includes: conducting decoloration on a ramoplanin fermentation solution through macroporous decolorization resin, then carrying out macroporous resin adsorption, desorption as well as concentration so as to obtain a ramoplanin crude extract containing the three single components: A1, A2, and A3; and dissolving the crude extract with a solvent, then performing chromatographic separation by means of a polymer microsphere, thus obtaining the products of three high purity ramoplanin single components A1, A2 and A3. The invention has the advantage that when using the polymer microsphere to perform chromatographic separation on the product, the sample treatment amount is large, and the obtained single components have high purity. As the price of a polymer microsphere medium is 30% of that of the reverse phase silica gel C18, and the separation effect is equivalent, the preparation cost is greatly reduced.

The invention provides a high-yield ansamitocin strain for reinforcing the polyketide synthase gene transcriptional level and a preparation method thereof, belonging to the technical field of biological medicine. The yield of ansamitocin can be increased by improving the rate-limiting enzyme gene expression level in biosynthetic pathway. In the rare actinosynnema pretiosum ATCC31280, the expression level of PKS gene ansa9 is reinforced by utilizing erythrocin resistance gene promoter, the biological synthesizing capacity of actinosynnema pretiosum can be reinforced, and the yield of actinosynnema pretiosum can be increased. The ansamitocin fermenting final yield of engineering strains is increased by about 125 percent in comparison with that of the original strain, and the lab shaking flask level reaches 99mg/L.

The invention discloses a method for enhancing positive regulatory protein gene expression to improve the acarbose fermentation level. In actinoplanes, positive regulatory protein genes ACPL_1889, ACPL_4236, ACPL_7303, ACPL_6479 and ACPL_8104 are respectively subjected to intensified expression by utilizing a strong promoter kasOp*, so that an acarbose high-yield mutant strain is obtained. Enhancement on the expression of the positive regulation protein genes can promote the expression of acarbose biosynthetic genes, and finally, obviously improve the acarbose yield. Compared with an originalstrain, the fermentation yields of the high-yield strains obtained by the invention are respectively improved by 25%, 18%, 26%, 22% and 15%, and the laboratory shake flask fermentation levels respectively reach 3.29g/L, 3.11g/L, 3.32g/L, 3.21g/L and 3.03g/L.

Synthesis and characterization of novel DACT forms suitable for pharmaceutical compositions in drug delivery systems to treat cancer in humans or a warm-blooded mammals. The novel forms include but not limited to cocrystals, salts, solvates of salts, and mixtures thereof.

The invention discloses a sponge-derived actinomycete and a preparation method and application of novel sulfur-containing alkaloid produced by the sponge-derived actinomycete. According to the present invention, a biological activity test results show that a compound 1 and a compound 2, which are separated from the fermentation product of the sponge-derived actinomycete Nocardiopsis dassonvillei SCSIO 40065, have antibacterial and antitumor activity, such that the compound 1 and the compound 2 can be used for the preparation of antibacterial drugs and antitumor drugs.