66 results about "Mycobacterium sp" patented technology

Fermentation of phytosterol compositions to produce androstenedione (androstene-3,17-dione) and subsequently androstenedione (androstene-1,4-diene,3,17-dione) A method comprising: propagating a microbial culture of the genus Mycobacterium in a first nutrient medium; placing the microbial culture and the phytosterol composition in a bioreactor for a sufficient time to convert the composition substantially to androgen enedione ("AD solution"); propagation of a fungal culture of Fusarium in a second nutrient medium; infusion of one or more of 1) Fusarium species or 2) fungal culture medium into the organism In the reactor, react for sufficient time to convert the AD solution to androsdienedione.

The invention provides a strain of Mycobacterium sp.16F for efficiently degrading polycyclic aromatic hydrocarbon and benzene organic matters, which has a preservation number of CGMCC No.6367. The mycobacterium 16F can efficiently, safely and rapidly degrade polycyclic aromatic hydrocarbons and benzene organic matters, can grow and degrade by using fluorene, naphthalene, anthracene, acenaphthene, phenanthrene, pyrene and benzopyrene as the sole carbon source and energy in aerobic condition, and can utilize benzene, m-xylene, toluene, salicylic acid, catechol and other multiple aromatic organic matters. The mycobacterium 16F is sensitive to streptomycin, rifampin, tetracycline, kanamycins and other antibiotics, has good degradation effects to mixed polycyclic aromatic hydrocarbons in aging soils and monocyclic benzene organic matters in water bodies, can be used for restoring and purifying the water-soil environment combinedly polluted by aromatic hydrocarbon organic matters, is important for promoting sustainable development, and has a wide application prospect.

The invention relates to a rehabilitation technology of polycyclic aromatic hydrocarbon pollutants (PAHs), particularly discloses a mixed inoculant for degrading PAHs. The mixed inoculant comprises indigenous mixed bacteria, wherein the indigenous mixed bacteria comprise Pseudomonas sp. B08, Pseudomonas sp. BM, Bacillus sp. BYB, Bacillus sp. B07, Ochrobactrum sp. ZHL4, Gordonia sp. BGDS and Mycobacterium sp. BFZG. The mixed inoculant disclosed herein can be added according to an inoculation quantity of 5-10%, and when the initial concentration of PAHs is 180.67mgL<-1>, the total degradation rate of PAHs after being processed for 9 days can reach 91.21%.

The invention relates to a petroleum degrading bacterium and an application thereof in crude oil degradation. The petroleum degrading bacterium is named as Mycobacterium sp.CSC-6, and is a Gram-negative bacterium. The bacterial strain is preserved in China Center for Type Culture Collection (CCTCC) in Nov. 27th, 2017. The preservation number is CCTCC No. M2017726. The bacterial strain is in a rodshape, has a light yellow color, and is opaque. After 48 hour culture, the diameter of the bacterial colony is 9.0-11.0 mm. The log-in number of the 16Sr DNA of the bacterial strain in GeneBank is MG490974.1. Compared with the prior art, the provided bacterial strain has high degradation efficiency, can well degrade short chain hydrocarbons, middle chain hydrocarbons, and long chain hydrocarbons,and is capable of completely degrading middle chain hydrocarbons and long chain hydrocarbons in crude oil, and at the same time, the degradation speed is quick.

The invention discloses a method for simultaneously removing USEPA PAHs in plant bodies by utilizing compound PAHs degrading bacteria. Multiple functional bacteria are prepared into a microbial compound functional fungicide, a plant absorption promoter, cytokinin 6-BA or KT, auxin 2,4-D and cysteine are added to prepare a degradation agent, then the degradation agent is inoculated in the plant bodies, and the PAHs content in the plant body is reduced; the compound fungicide is prepared from 8 PAHs degrading bacteria different in degradation spectrum, wherein the degrading bacteria are Sphingobiumsp sp. RS1 and RS2, mycobacterium sp. Pyr9, 033, Diaphorobacter sp. Phe15 and Massilia sp. Pn2, Paenibacillus sp. Phe3, Pseudomonas sp. Ph6 respectively. The acenaphthylene, acenaphthene, fluorene, phenanthrene, anthracene, fluoranthene, pyrene, benzo[a] anthracene, benzo[b]fluoranthene, benzo[k]fluoranthene, benzo[a]pyrene, dibenzo[a,h] anthracene, benzo[ghi]indene and [1,2,3-cd] pyrene in each plant body in a contaminated area can be effectively removed and degraded, the removal rate of PAHs is high, and the method has the advantages of high efficiency, environmental protection and easy operation.

The invention provides a method for preparing androstenedione using a double liquid phase fermentation system by fermenting Mycobacterium sp.DE6823, and aseptic distilled water is selected as a fermentation medium water phase, and rice oil, sunflower oil, soybean oil and peanut oil can be selected as an oil phase, after fermentation conversion, a solution containing androstenedione is obtained, and after condensation, decolouring, filtering, condensation, recrystallization and vacuum drying and other operations, and androstenedione whose purity is above 99% is obtained. Different oil phases can be selected, and the conversion rate of the plant sterol is in the range of 59%-85%. The invention solves the problems that the raw materials of the present androstenedione production technology in our country are limited by season and region, thereby effectively reducing destroy to ecological environment; the invention has the characteristics of simple operation, small environmental pollution, high conversion rate, low production cost, etc., and is suitable for industrialized production.

The invention discloses a bacterium capable of degrading herbicide acetochlor and an application of the bacterium. The bacterium is mycobacterium sp. YCA11 and the preservation number of the bacterium is CGMCC (China General Microbiological Culture Collection Center) No. 7771. The mycobacterium sp. YCA11 with the preservation of CGMCC No.7771, disclosed by the invention, has the advantage that the degradation rate of the bacterium to 45mg/L acetochlor is up to 51.86% in 7 days in an inorganic salt culture medium, which indicates that the bacterium is capable of efficiently degrading the acetochlor and has a broad application prospect in the remediation of soil polluted by the acetochlor.

The invention provides five pairs of primers and an identification method for PCR(Polymerase Chain Reaction) identification of mycobacterium bovis. The five pairs of primers respectively amplify aiming at a 16SrRNA conserved region of mycobacterium bacteria, a mycobacterium tuberculosis complex(MTBC)Rv0577 gene, Rv1970 of a mycobacterium tuberculosis RD7 region, a mycobacterium bovis pncA gene and an RD1 region gene to generate specific amplified fragments, and the nucleotide sequences of the specific amplified fragments are as shown in SEQ ID No.1-5. According to the identification method provided by the invention, the total DNA of a sample is taken as a template, PCR amplification is respectively performed by the five pairs of primers, and the result is judged according to the size of an amplified band. The primers provided by the invention can specifically identify mycobacteria, MTBC, mycobacterium tuberculosis, mycobacterium bovis and mycobacterium bovis BCG(Bacillus Calmette-Guerin), and the detection method has good sensitivity and simplicity of method and operation, and can realize quick large-flux detection of mycobacterium bovis.

The invention provides a method for preparing androstenedione using a double liquid phase fermentation system by fermenting mycobacterium sp.DE6823, and aseptic distilled water is selected as a fermentation medium water phase, and rice oil, sunflower oil, soybean oil and peanut oil can be selected as an oil phase, after fermentation conversion, a solution containing androstenedione is obtained by employing a membrane separating method, and after condensation, decolouring, filtering, condensation, recrystallization and vacuum drying and other operations, androstenedione with purity of 99% or more is obtained. Different oil phases can be selected, and the conversion rate of the plant sterol is in the range of 59%-85%. The invention solves the problems that the raw materials of the present androstenedione production technology in our country are limited by season and region, thereby effectively reducing destroy to ecological environment; and the invention has the characteristics of simple operation, small environmental pollution, high conversion rate, low production cost, etc., and is suitable for industrialized production.

The invention belongs to the technical field of biology, and particularly relates to mycobacterium sp. LY-1 capable of efficiently converting phytosterol and application thereof. The strain is preserved in the common microorganism center of China Committee for Culture Collection of Microorganisms, and the preservation number is CGMCC No.13031. The mycobacterium sp. LY-1 is capable of converting phytosterol into 9 alpha-hydroxy androstane-4-alkenyl-3,17-diketone, and the product yield is 33% under the condition that the substrate feed amount is 15 g/L.

The invention discloses high-efficiency organic pollutant carbendazim degrading bacteria and use thereof. The organic pollutant carbendazim degrading bacteria are (mycobacterium sp.)DJL22, and were collection in China General Microbiological Culture Collection Center, and the collection number of organic pollutant carbendazim degrading bacteria is CGMCC No.5041. When the (mycobacterium sp.)DJL22 CGMCC No.5041 is cultured in inorganic salt culture medium for 5 days, the rate of degradation of carbendazim at a concentration of 100mg/L reaches 84.79 percent; and when the (mycobacterium sp.)DJL22CGMCC No.5041 is cultured in inorganic salt culture medium for 15 days, the rate of degradation of carbendazim at a concentration of 600mg/L reaches 62.77 percent. Therefore, the stain can efficiently degrade carbendazim and has a bright application prospect in repairing soil polluted by carbendazim.

The present invention relates to the production of a product compound having a structure according to Formulae IA and/or IB:

wherein n is 0 or 1;

• • • R¹ is hydrogen or hydroxy;
    • R² is hydrogen;
    • or, when n is 0, R¹ and R² taken together form a second bond between the carbon atoms bearing R¹ and R², provided that when n is 1, R¹ and R² are each hydrogen;
    • R³ is —COOH or —COOR⁴;
    • R⁴ is an alkyl or aryl moiety; A, B, and D are the substituents of their rings, each of which may be different or the same, and are selected from the group consisting of hydrogen, halogens, alkyl, hydroxy, and alkoxy.

This process involves incubating a starting compound having a structure according to Formulae IIA and/or IIB:

wherein R³ is —CH₃ and R¹, R², A, B, and D are defined above.

in the presence of a microorganism under conditions effective to produce the product compound. The microorganism can be from the genus Streptomyces, Stemphylium, Gliocladium, Bacillus, Botrytis, Cyathus, Rhizopus, Pycniodosphora, Pseudomonas, Helicostylum, Aspergillus, Mucor, Gelasinospora, Rhodotorula, Candida, Mycobacterium, or Penicillium. Alternatively, the microorganism can be Cunninghamella bainieri.

The invention discloses a detection method for identifying mycobacterium tuberculosis and non-mycobacterium tuberculosis infections. The detection method comprises the following steps: respectively contacting the specific antigen of mycobacterium tuberculosis and the specific antigen of mycobacterium with a to-be-detected in-vitro T cell, and detecting a cell factor released by the T cell; comparing the detection results of the specific antigen of mycobacterium tuberculosis and the specific antigen of mycobacterium; judging that the to-be-detected in-vitro T cell is suffered from mycobacterium tuberculosis infection or non-mycobacterium tuberculosis infection according to a comparison result. According to the detection method provided by the invention, the specific antigen of mycobacterium and the ELISPOT detection method for detecting the specificity of MTB can be combined and utilized to simultaneously identify MTB and NTM infections. The detection method provided by the invention has important guidance significance on clinical diagnosis and drug usage of a doctor.

An object of the present invention is to provide an oligonucleotide for rapidly and conveniently detecting bacteria of the genus Mycobacterium (acid-fast bacteria) or for identifying the bacterial species thereof, and a method and kit for detecting bacteria of the genus Mycobacterium (acid-fast bacteria) using such oligonucleotid. The present invention provides a method for identifying Mycobacterium tuberculosis, which comprises performing a nucleic acid amplification reaction using a primer for nucleic acid amplification that comprises a nucleotide sequence corresponding to a variable region in a 16S rRNA gene sequence of Mycobacterium tuberculosis and has at least 3 continuous nucleotides contained in the nucleotide sequence represented by SEQ ID NO: 1 at the 3′ end.

The invention discloses a mycobacterium and application thereof. The mycobacterium is classified and named as mycobacterium JYC-07, and is preserved in China Center for Type Culture Collection on June9th, 2017, and the preservation number is CCTCC NO : M 2017325. The strain Mycobacterium sp.JYC capable of converting canrenone into 9alpha-hydroxycarvenone by microorganisms is obtained by screeningand separating from a natural environment, the strain Mycobacterium sp.JYC-07 capable of producing 3-sterone-9alpha hydroxylase at high yield is further obtained through genetic engineering, the strain can efficiently convert canrenone into 9alpha-hydroxycarvenone, the conversion rate is over 90 percent, and the strain has very important significance for expanding the application of steroid raw material drug canrenone and good industrial prospects.

Oligonucleotides used to prime in vitro nucleic acid amplification of 16S rRNA sequences or DNA encoding 16S rRNA sequences for many species within the genus Mycobacterium are disclosed. Kits including such oligonucleotides are disclosed. Methods of detecting Mycobacterium species using the oligonucleotides in in vitro nucleic acid amplification are disclosed.

The present invention discloses a high-androstenedione-producing recombinant mycobacterium and a construction method and an application of the high-androstenedione-producing recombinant mycobacterium.The construction method is as follows: cholesterol oxidase gene ChoM2, sterone C27 monooxygenase gene Smo2 carrying an RBS fragment and 17[beta]-hydroxysterol dehydrogenase gene Hsd4A carrying an RBSfragment are introduced into mycobacterium sp. CICC 21097 to obtain the high-androstenedione-producing recombinant mycobacterium MNCSH. A fermentation main product of the recombinant strain obtainedby the method is 4-androstenedione, the yield is 90% or more, the selectivity is 90% or more, formation of by-products is reduced and the yield of the androstenedione is increased. Compared with an original strain, the recombinant strain has better catalytic conversion performance and can improve production efficiency, and the processing method is simple and efficient.

The present invention belongs to the field of microbial gene detection and particularly relates to a film strip for identifying mycobacterium tuberculosis, mycobacterium chelonae, mycobacterium abscessus, and preparation and use methods of the film strip. Specific probes comprise base sequences for detecting the mycobacterium tuberculosis in SEQ ID NO.1-2; base sequences for detecting the mycobacterium chelonae in SEQ ID NO.3-4; base sequences for detecting the mycobacterium abscessus in SEQ ID NO.5-6 and mycobacterium quality control probes SEQ ID NO.7-8; and corresponding primer pairs are SEQ ID NO.9-10. The established detection method can rapidly and accurately distinguish the mycobacterium tuberculosis, mycobacterium chelonae and mycobacterium abscessus, can be used as a useful supplementary identification method for a commercialized mycobacterium strain identification kit, and has good clinical application prospects.

Disclosed are a primer composition including two pairs of primers targeting one gene (mpb64) concerning Mycobacterium sp., and a pair of primers targeting a human-derived gene (HLA-DR); and a method for detecting Mycobacterium sp. using a multiplex one-tube nested PCR method in which their primers are used to amplify two genes in one tube at the same time. The method of the present invention may provide a useful guide post capable of clinically diagnosing Mycobacterium sp. in a specimen in a more specific, rapid and convenient manner.

The invention discloses a 17[beta]-hydroxysteroid dehydrogenase mutant of mycobacteria and heterologous expression of the mutant, and belongs to the field of enzyme gene engineering and enzyme engineering. The invention provides actinomycete 17[beta]-hydroxysteroid dehydrogenase 17[beta]-HSDy1 from mycobacterium sp. LY-1 and a gene sequence of the 17[beta]-HSDy1. By taking pMA5 as an expression plasmid and bacillus subtilis WB600 as an expression host, heterogenous expression of the 17[beta]-HSDy1 of the mycobacterium sp. LY-1 is realized. A key T239 site of the dehydrogenase obtained throughsoftware simulation is mutated into tyrosine, and the enzyme activity is improved by 19.2% and reaches 9571.77 U.mg<1>. An enzyme activity-growth curve of recombinant bacillus subtilis WB600-pMA5-17[beta]HSDy1-T239Y obtained by mutation is measured, and the specific enzyme activity of the recombinant bacillus subtilis WB600-pMA5-17[beta]HSDy1-T239Y reaches the maximum after fermentation is performed for 28 hours. The heterologously expressed strain can be used for production of androstane-1,4-ene-3,17-dione converted boldenone, and has potential industrial application.

The invention discloses a method for transforming and producing 9-OH-AD by using whole cells of recombinant corynebacterium glutamicum, and belongs to the technical field of bioengineering and biology. Through a shuttle plasmid pXMJ19 between escherichia coli and corynebacterium glutamicum, an oxidization subunit KshA and a reducing subunit KshB of 3-ketosteroid 9alpha-hydroxylase from Mycobacterium sp.Strain VKM Ac-1817D are successfully co-expressed in C.glutamicum ATCC13032. A whole-cell thallus of the engineering bacterium is used as a biocatalyst to transform and produce 9alpha-hydroxyl androstane-4-alkene-3,17-diketone; the mole transformation rate of the substrate androstane-4-alkene-3,17-diketone is up to 99.5 percent. The 9alpha-hydroxyl androstane-4-alkene-3,17-diketone producedby the method disclosed by the invention has the advantages of high efficiency, high specifity, low energy consumption and the like.