48 results about "Aspergillus ochraceus" patented technology

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 provides a method for catalyst production of 11alpha-hydroxy-16alpha, 17alpha-epoxyprogesterone in a binary system by the use of aspergillus ochraceus. By the addition of an organic solvent dibutyl phthalate, dissolving problem of a substrate and a product is effectively solved, contact area with cells is increased, separation process is simplified, conversion time is shortened, conversion rate of the substrate is greatly raised, and conversion rate of 16 alpha, 17 alpha-epoxyprogesterone can reach more than 84%. The method provided by the invention lays a solid practical production foundation for production of high-yield and high-purity 11 alpha-hydroxy-16 alpha, 17 alpha-epoxyprogesterone. Production costs are reduced, and economic benefits of enterprises are raised.

The invention discloses a method for preparing 11Alpha,17Alpha-hydroxyprogesterone, comprising: using 17Alpha-hydroxyprogesterone as a substrate, adding magnetic nano ferroferric oxide powder into a culture medium, and preparing 11Alpha,17Alpha-hydroxyprogesterone by Aspergillus ochraceus fermentation process; the method has the advantages that feeding concentration is high, substrate molar conversion rate is high, the feeding concentration is up to 30 g / L, and the highest molar conversion ratio is up to 91.9%; the magnetic nano ferroferric oxide powder is recyclable; by using the magnetic nano ferroferric oxide powder, it is possible to greatly increase hydroxyl conversion rate, the process is simple, the production cost is simple, and the method has great industrial application prospect.

The invention provides bacillus subtilis. The collection number of the bacillus subtilis is CCTCC NO:M2017708. The invention further provides application of the bacillus subtilis to degradation of ochratoxin A (OTA) in post-fermented tea pile fermentation. The bacillus subtilis can adapt to a post-fermented tea pile fermentation environment (acidic, high temperature and anaerobic), and has the capability of degrading the ochratoxin A; after the ochratoxin is fermented and degraded through the collection bacteria, the main quality component of a fermented tea sample changes slightly; the drinking safety of fermented tea is ensured by degradation of the ochratoxin A generated in a tea pile fermentation process with the bacteria.

The present invention relates to anti-saline-alkali aspergillus ochratensis strain W1 and a micro-organism agent thereof, and applications of the anti-saline-alkali aspergillus ochratensis strain W1.The strain is aspergillus ochratensis preserved in China General Microbiological Culture Collection Center and has a preservation name of aspergillus ochratensis W1 and a preservation number of CGMCCNO.17971. The anti-saline-alkali aspergillus ochratensis strain W1 can stably survive in the soil with high sodium salt concentration and high pH, effectively degrades straws and is suitable for rebuilding saline-alkali land.

The invention discloses a high-expression cytochrome P450 monooxygenase aspergillus ochraceus strain, a construction method thereof and application thereof in fermentation production of 11alpha-hydroxy-canrenone. The aspergillus ochraceus strain is currently preserved in China Center for Type Culture Collection with the preservation number of CCTCC NO: M2019616, wherein the recombinant fragment comprises a cytochrome P450 monooxygenase coding gene, upstream and downstream DNA sequences, a promoter, a terminator and an antibiotic resistance gene; and homologous recombination occurs between thegene fragment and the cytochrome P450 monooxygenase gene in the chromosome of an aspergillus ochraceus initial strain MF018. according to the invention, a homologous recombinant aspergillus ochraticastrain is obtained by the double exchange which occurs between the constructed high-expression cytochrome P450 monooxygenase gene and the cytochrome P450 monooxygenase gene in the chromosome of the aspergillus ochraceus initial strain MF018. The invention can realize the effective accumulation of 11alpha-hydroxy-canrenone produced in the fermentation process.

The invention relates to a method for extracting a streptomyces antibacterial product, and belongs to the technical field of biology. Streptomyces sp S24 is activated and fermented; the fermentation solution is sterilized, centrifuged, absorbed by macroporous absorption resin AB-8 and desorbed by 75 to 85 percent acetone; and a solvent is reclaimed from the desorption solution, and the desorptionsolution is evaporated and dried to obtain an antibacterial product extract. Experiments prove that the antibacterial product has broad-spectrum antibacterial activity for main mildewing fungi such as aspergillus flavus, aspergillus niger, aspergillus ochraceus and aspergillus fumigatus in food and feed and human pathogenic fungi such as aspergillus flavus and aspergillus fumigatus. Because the antibacterial product provided by the invention has obvious inhibiting effect on various food and geed mildewing fungi and human pathogenic fungi, the antibacterial product shows efficient broad-spectrum properties; and the method for extracting the antibacterial product is simple and convenient, has low cost, and has high practical application value and broad market prospect on the aspects of foodpreservative, feed additive and medicaments.

The invention discloses a method for preparing 16alpha, 17alpha-epoxy-11alpha-hydroxy-pregna-1,4-diene-3,20-dione by taking 16alpha,17alpha-epoxy progesterone as a starting material through one-step mixed fermentation of aspergillus ochraceus and arthrobacter simplex.

The invention discloses cinnamyl aldehyde Pickering emulsion for inhibiting growth and toxin production of aspergillus ochraceus and a preparation method thereof. In the emulsion, the content of cinnamylaldehyde is 5 to 30 g / L, the content of chitosan is 0.125 to 0.625 g / L, the content of corn protein is 2.5 to 12.5 g / L, the content of ethanol is 125 to 250 mL / L, and the content of acetic acid is 0.5 to 1.25 mL / L. Cinnamaldehyde biological resources are widely sourced in China, the production process is simple, the cost is low, and the cinnamaldehyde has volatility; the emulsion has good safety to human and livestock, and has been listed in a food additive directory (GB 2760-2014 national food safety standard - standard for use of food additives); the emulsion is novel in thought, the emulsion preparation method is simple, rapid and efficient, and the prepared cinnamylaldehyde Pickering emulsion can keep good stability in a long time and is high in practicability; and solid particles (the chitosan and the corn protein) serving as emulsifiers are wide in source, safe, non-toxic and degradable. The cinnamylaldehyde Pickering emulsion prepared by the invention has a good effect of inhibiting the growth and toxin production of the aspergillus ochraceus, and can be directly applied to mildew-proof preservation of grains.

The present invention relates to a novel cytochrome P450-like enzyme (Aspergillus ochraceus 11 alpha hydroxylase) and an oxidoreductase (Aspergillus ochraceus oxidoreductase) isolated from cDNA library generated from the mRNA of Aspergillus ochraceus spores. When the cDNA encoding the 11 alpha hydroxylase was co-expressed in Spodoptera frugiperda (Sf-9) insect cells with the cDNA encoding human oxidoreductase as an electron donor, it successfully catalyzed the conversion of the steroid substrate 4-androstene-3,17-dione (AD) to 11 alpha-hydroxy-AD as determined by HPLC analysis. The invention also relates to nucleic acid molecules associated with or derived from these cDNAs including complements, homologues and fragments thereof, and methods of using these nucleic acid molecules, to generate, for example, polypeptides and fragments thereof. The invention also relates to the generation of antibodies that recognizes the A. ochraceus 11 alpha hydroxylase and oxidoreductase and methods of using these antibodies to detect the presence of these native and recombinant polypeptides within unmodified and transformed host cells, respectively. The invention also provides methods of expressing the Aspergillus 11 alpha hydroxylase gene separately, or in combination with human or Aspergillus oxidoreductase, in heterologous host cells, to facilitate the bioconversion of steroid substrates to their 11 alpha hydroxy-counterparts.

The invention relates to actinomycetes for resisting contamination fungi in grain feeds and grains, which is characterized in that: the strain is named TD-1 and is classified and named Streptomyces sp., which is collected in China General Microbiological Culture Collection Center of Beijing on March 15, 2011 with the collection number of CGMCC No.4666. Autotrophic actinomyces for resisting growth of contamination fungi in the grain feeds and grains is separated and screened for the first time; and the strain has obvious inhibition effect on various fungi contaminating the grain feeds and grains, has obvious antagonism on aspergillus flavus, penicillium citrinum, fusarinm solani, aspergillus ochraceus, aspergillus oryzae, aspergillus nidulans, aspergillus niger, fusarium oxysporum, penicillium viridicatum and the like, can effectively prolong the retention period and improve the storing quality of the grain feeds and the grains, and lays a foundation for developing biological inhibitors for the fungi in the grain feeds and the grains.

The invention relates to an ochratoxin A aptamer affinity column and a preparation method and application thereof. The affinity purification column utilizes a chemically modified solid phase carrier, and then a nucleic acid aptamer of ochratoxin A is covalently coupled to the carrier. Then pack the affinity column. The aptamer affinity column is mainly used for the purification of ochratoxin A in food, feed, milk, blood samples and other various samples, so as to facilitate the high performance liquid chromatography (HPLC) of ochratoxin A in the sample in the later stage detection and fluorescence detection.

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 discloses cinnamaldehyde-containing emulsion for inhibiting the growth of aspergillus ochraceus and a preparation method thereof, the emulsion system is prepared from cinnamaldehyde, an emulsifier and water, the emulsifier is composite particles prepared from chitosan and corn protein through an ultrasonic method, the content of cinnamaldehyde in the emulsion is 5-30g / L, the content of chitosan is 0.5-2.5g / L, and the content of corn protein is 5-25g / L. Cinnamaldehyde biological resources are widely sourced in China, the production process is simple, the cost is low, and the cinnamaldehyde has volatility; cinnamaldehyde is good in safety to people and livestock and is listed in a food additive catalog (GB 2760-2014); the method is novel in thought, the emulsion preparation method is simple, and the prepared cinnamaldehyde-containing emulsion can keep good stability within 30 days and is high in practicability; the prepared cinnamaldehyde-containing emulsion has a good effect of inhibiting aspergillus ochraceus, and is safe and free of toxic and side effects.

The invention discloses a method for preparing 11alpha, 17alpha-hydroxyprogesterone by utilizing immobilized hydroxylase conversion, which is characterized in that 17alpha-hydroxyprogesterone is used as a substrate, the 11alpha, 17alpha -hydroxyprogesterone is prepared by utilizing the immobilized hydroxylase conversion, and the method comprises the following steps of: preparing a culture medium, culturing aspergillus ochraceus in the culture medium, inducing hydroxylase, preparing the immobilized hydroxylase, preparing a conversion system, converting a substrate, extracting and refining a product and the like. The method has the advantages that the feeding concentration is high, the substrate conversion rate is high, the conversion period is short, the product is easy to separate, and the immobilized hydroxylase can be repeatedly used. The feeding concentration of 17 alpha-hydroxyprogesterone in a single enzyme reaction is 40 g / L, the molar conversion rate can reach 93%, and meanwhile, the immobilized hydroxylase can be repeatedly recycled for more than three times. The preparation method is simple in operation process and relatively low in production cost, and has a relatively high industrial application prospect.

The invention provides a preparing method of an 11alpha-hydroxy-16alpha, 17alpha-epoxy progesterone dehydrogenation substance. The preparing method comprises the steps of conducting methylbenzene-resistant aspergillus ochraceus microbial catalytic treatment on 11alpha-hydroxy-16alpha, 17alpha-epoxy progesterone in a two-phase fermentation solution which uses sterile water as an aqueous phase and methylbenzene as an organic phase, and obtaining the 11alpha-hydroxy-16alpha, 17alpha-epoxy progesterone dehydrogenation substance. According to the preparing method of the 11alpha-hydroxy-16alpha, 17alpha-epoxy progesterone dehydrogenation substance, by dissolving 11alpha-hydroxy-16alpha, 17alpha-epoxy progesterone in a molecular state into a methylbenzene solution, the contact area of 11alpha-hydroxy-16alpha, 17alpha-epoxy progesterone and biological enzyme in methylbenzene-resistant aspergillus ochraceus cells is increased, and thus time for conversion is shortened, and efficiency of a C1,2 dehydrogenation reaction conducted on 11alpha-hydroxy-16alpha, 17alpha-epoxy progesterone is sharply increased. The preparing method of the 11alpha-hydroxy-16alpha, 17alpha-epoxy progesterone dehydrogenation substance is simple in process and easy for industrialization production.

The present invention relates to a process for the detection of Aspergillus ochraceus by biotechnological approach. This invention particularly relates to the development of a process for the detection of species belonging to Aspergillus ochraceus group by PCR / multiplex PCR technique. More particularly the present invention relates to a set of novel oligonucleotide primers and the method for the detection of Aspergillus ochraceus thereby.

The invention provides an aspergillus ochraceus steroid C11[alpha]-hydroxylase mutant I303L, wherein the DNA sequence of the mutant I303L is shown as SEQ ID NO:2. Compared with C11[alpha]-hydroxylase CYP68J5, the mutant I303L shows higher specificity when being used for converting a steroid substrate progesterone, and no obvious by-product is generated. The invention also provides a heterologous overexpression expression vector based on the mutant I303L, a pichia pastoris recombinant strain and a steroid transformation process of the pichia pastoris recombinant strain, and valuable materials (genes and strains) and basic data support are provided for research and development of an efficient progesterone C11[alpha]-hydroxylation production process.

The invention relates to a primer and a probe for simultaneously detecting aspergillus flavus and aspergillus ochraceus. The invention further relates to a double microdroplet digital PCR detection method for determining the aspergillus flavus and the aspergillus ochraceus in the food. The method comprises the step of using the specific oligonucleotide primer and the probe for detecting the aspergillus flavus and the aspergillus ochraceus. The invention further relates to a double microdroplet digital PCR detection kit for the aspergillus flavus and the aspergillus ochraceus in the food. The kit comprises the specific oligonucleotide primer and the probe. By using the double microdroplet digital PCR detection method, the aspergillus flavus and the aspergillus ochraceus can be simply, quickly, specifically and sensitively detected at the same time.

The invention discloses a method for preparing delta 1-11 alpha,17 alpha-dihydroxyprogesterone with a one-pot continuous fermentation process, and belongs to the technical field of intermediate preparation. The method comprises the following specific steps: (1) inoculating slant culture of arthrobacter simplex into a first culture medium, performing culture for 20-30 hours, adding ground 17 alpha-hydroxyprogesterone, and performing transformation; (2) inoculating fermentation liquor after finishing the step (1) into a second culture medium, inoculating slant culture of aspergillus ochraceus into the second culture medium, performing culture, and performing post-treatment to obtain delta 1-11 alpha,17 alpha-dihydroxyprogesterone. Delta 1-11 alpha,17 alpha-dihydroxyprogesterone is prepared with the one-pot continuous fermentation process, so that a step of filtering, extracting, refining and drying a dehydrogenation material is reduced, discharge of 50% pollutants is reduced, environmental pressure is reduced, the time is saved, the yield is increased, the efficiency is improved, the process is simplified, the whole reaction operation is convenient, and the method is suitable for industrialized popularization and application.

The invention relates to a mildew-proof biological fermentation feed and a preparation method thereof, and the mildew-proof biological fermentation feed comprises the following raw material components in parts by weight: 100-150 parts of a coarse feed, 2-4 parts of an antagonistic microorganism fermentation product and 10-20 parts of a compound fermentation strain, wherein the antagonistic microorganism fermentation product is composed of freeze-dried powder of a fermentation product of the Bickholderia and freeze-dried powder of a fermentation product of the streptomycete, and the freeze-dried powder of the fermentation product of the Bickholderia and the streptomycete is added into the fermentation raw materials in the application. The activity of aspergillus flavus and aspergillus ochraceus possibly introduced in the fermentation and storage processes of the fermented feed is inhibited, so that the mildew-proof effect of the fermented feed is improved. In order to further enhance the mildewproof effect, two fermentation products of sansevieria trifasciata prain and basil leaves are added into the fermentation raw materials, so that the mildewproof effect of the fermented feed is further improved due to the synergistic effect of the two fermentation products and the microbial fermentation product freeze-dried powder.

The invention discloses a crude polysaccharide detection method. The crude polysaccharide detection method comprises the following steps: a. carrying out solid inclined plane culture on aspergillus ochraceus seeds; b. filtering a fermentation solution; c. making the volume to be constant by de-ionized water, and sufficiently dissolving and uniformly mixing to be used as a test sample solution for later use; d. preparing 1mg / ml standard sample water solution for later use; e. preparing a thin-layer chromatography plate and an unfolding chamber; f. measuring 2 microliters of a monosaccharide standard product water solution and 2 microliters of a crude polysaccharide testing solution; sequentially carrying out sample application on a sample application line of the thin-layer plate; and g. unfolding an unfolding agent to the part which is 2cm far away from the upper end of the thin-layer plate, taking out the thin-layer plate, airing and spraying an aniline-phthalic acid solution, immediately drying by blowing and heating at 110 DEG C until spot color development is clear so as to observe a detection result of the crude polysaccharide. According to the detection method, a thin-layer chromatography method is used for determining the content of the crude polysaccharide in the fermentation solution and mycelia, and the detection result can be rapidly and accurately obtained; and the operation is simple and convenient and the method is particularly suitable for a qualitative analysis research of a sample.

The present invention relates to a saline-alkali-resistant Ochraus ochraceus strain W1 and its agent and application thereof. The strain is Aspergillus ochraceus preserved in the General Microorganism Center of China Microbiological Culture Preservation Management Committee, and the preserved name is Ochraus ochraceus W1 , the deposit number is CGMCC NO.17971. The saline-alkali-resistant ochrax strain W1 can survive stably in soil with high sodium concentration and high pH, ​​effectively degrades straw, and is suitable for transforming saline-alkali land.

The invention provides a novel inducible promoter from filamentous fungus aspergillus niger and plasmid vector construction thereof, wherein the amino acid sequence of the novel inducible promoter is as shown in SEQ ID NO:1; the promoter can induce efficient inducible expression of exogenous genes in aspergillus niger cells, such as high-level expression of P450 steroid hydroxylase genes; the high-level expression of the aspergillus ochraceus C11[alpha]-hydroxylase gene 68J5 in aspergillus niger ATCC1015 cells and the conversion efficiency of recombinant bacteria to steroid substrate 16,17[alpha]-epoxy progesterone are investigated, so the aspergillus ochraceus C11[alpha]-hydroxylase gene 68J5 can be used for constructing aspergillus niger high-efficiency gene engineering bacteria.

The invention discloses a method for determining the content of ochratoxin A in fruit juice. The samples were pretreated by a simple liquid-liquid extraction purification step, and then scanned and collected the three-dimensional fluorescence data of the standard and samples under the optimized scanning wavelength and scanning interval parameters, and the obtained data were analyzed by parallel factor analysis (PARAFAC) Carry out mathematical separation processing, use "mathematical separation" to combine chemical and physical separation, use known concentration standards to establish a calibration model, and realize the prediction of the components to be measured in the case of unknown, uncorrected background interference and serious spectral overlap . The method is simple, rapid, and highly sensitive, and can realize the determination of ochratoxin content in fruit juice under the interference of unknown background. It belongs to the field of food safety.

The invention belongs to the technical field of sterol biotransformation, and in particular, relates to a method for preparing a steroid drug intermediate 11[alpha]-hydroxyandrost-4-en-3,17-dione by mixed bacterial fermentation biotransformation of phytosterol. A fermentation culture medium containing phytosterol and a transformation medium cyclodextrin is inoculated with a mycobacterium TCCC 11028 M3, culture is performed for 24 h, then the fermentation culture medium is inoculated with an aspergillus ochraceus CICC 41473 spore solution, and transformation continues to be carried out. One-step transformation from the phytosterol to 11[alpha]-hydroxyandrost-4-en-3,17-dione is achieved by mixed bacterial fermentation. The method improves the production rate of the product, shortens the transformation period, simplifies the operation, saves the cost, and is suitable for production of steroid compound biotransformation.

Relating to the field of removal of fungal toxins, the invention discloses a compound enzyme, an additive, application thereof and a method for removal of fungal toxins. Specifically, the invention relates to a compound enzyme, which contains amidase and esterase and can remove fungal toxins, especially fumonisins, ochratoxins and T2 toxin, additives containing the compound enzyme and applicationthereof in removal of fungal toxins, especially fumonisins, ochratoxins and T2 toxin, and a method for removal of fungal toxins. According to the technical scheme, combined use of the amidase and esterase can achieve simultaneous removal of ochratoxin A, fumonisins and T2 toxin, the removal efficiency is greatly improved than single use of one enzyme, and vomitoxins, aflatoxins and zearalenone toxin can be removed to certain degree.

The invention relates to lactobacillus rhamnosus capable of eliminating ochratoxin A. The lactobacillus rhamnosus is preserved in the China Center for Type Culture Collection on January 5, 2022, and the preservation number is CCTCC M 2022013. Lactobacillus rhamnosus is activated, cultured and centrifuged to obtain thalli, and the centrifuged thalli are washed and centrifuged to obtain the lactobacillus rhamnosus biological preparation. And resuspending the thalli by using a protective agent until the concentration reaches 1010 CUF / mL, culturing the suspension, and freeze-drying to obtain the lactobacillus rhamnosus powder biological preparation. The lactobacillus rhamnosus (Lactobacillus rhamnosus Bm01) provided by the invention has acid resistance, can be used for rapidly removing the ochratoxin A in an aqueous solution, can be used for remarkably reducing the ochratoxin A in grape juice and coffee, is used for removing the ochratoxin A in food, and has a very wide application prospect.

The invention discloses a compound of a targeted Keap1-Nrf2-ARE signal channel, a preparation method and application of the compound of the targeted Keap1-Nrf2-ARE signal channel. The compound is obtained by co-culturing penicillium and aspergillus ochraceus and extracting and separating secondary metabolites in fermentation liquor of the penicillium and the aspergillus ochraceus. The compound not only has no toxic effect on nerve cells, but also has a protective effect on nerve cells damaged by oxidation. Specifically, the compound plays a role in protecting nerve cells through a targeted Keap1-Nrf2-ARE signal channel, and the compound realizes a regulation effect on the signal channel through a non-covalent binding mode of forming a hydrogen bond and a part of hydrophobic bonds by binding Keap1, so that not only can activated release of Nrf2 be realized, but also the structure of Keap1 can be prevented from being damaged, and the compound has a good application prospect. Therefore, the safety of the compound in targeting a Keap1-Nrf2-ARE signal channel is improved.