Streptomyces violaceorubidus and application thereof
A technology of Streptomyces crimsonus, JG-1, which is applied in the direction of bacteria, enzymes, biochemical equipment and methods, etc., to achieve the effects of rapid growth, short fermentation cycle and wide application prospects
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Embodiment 1
[0027] Embodiment one: Streptomyces purpurea ( Streptomyces violaceorubidus ) Isolation and screening of JG-1 CGMCC No.7921
[0028] 1. Separation
[0029] Streptomyces crimsonus JG-1 of the present invention is isolated from Kyzyl-Suu (N 41 ° 18' 25.12 E 72 ° 57' 46.15) wild walnut forest soil samples in Jalalabad, Kyrgyzstan, and randomly selects 5 The soil samples at a depth of 3cm-20cm were mixed and put into sterile containers, transported to the laboratory in a car refrigerator, and a strain of Streptomyces JG-1 was isolated by flat plate dilution coating method.
[0030] Separation steps: According to the gradient dilution method, weigh 10g of soil samples in 90mL sterile physiological saline, and perform gradient dilution after activation at 30°C for 30min. Select 10 -2 、10 -3 、10 -4 The dilutions were spread on the plates of starch agar medium and glycerol-asparagine agar medium respectively, each treatment was replicated three times, and cultured at 30°C. Aft...
Embodiment 2
[0040] Embodiment two: Streptomyces purpurea ( Streptomyces violaceorubidus ) Sequence determination and phylogenetic analysis of JG-1 CGMCC No.7921
[0041] 1. DNA extraction: JG-1 genomic DNA was extracted by CTAB / NaCl method. Inoculate the strain in starch medium (soluble starch 20g, sodium chloride 0.5g, potassium nitrate 1g, dipotassium hydrogen phosphate 0.5g, magnesium sulfate heptahydrate 0.5g, ferrous sulfate heptahydrate 0.01g, dilute to 1L, pH 7.0-7.2, sterilized at 121°C for 20 minutes), cultured with shaking at 30°C overnight. Take 1.5ml culture and centrifuge at 12000rpm for 2min. Add 567 ul of TE buffer to the precipitate, pipette repeatedly to resuspend it, add 30 ul of 10% SDS and 15 ul of proteinase K, mix well, and incubate at 37°C for 1 h. Add 100ul 5mol / L NaCl, mix well, then add 80ul CTAB / NaCl solution, mix well, and then incubate at 65°C for 10min. Add an equal volume of phenol / chloroform / isoamyl alcohol and mix well, centrifuge for 4-5min, transf...
Embodiment 3
[0073] Embodiment three: Streptomyces crimson purple ( Streptomyces violaceorubidus ) JG-1 CGMCC No.7921 enzyme activity screening
[0074] Enzyme-producing strains were screened by the plate transparent circle method, and a total of 7 kinds of enzyme activities were screened: α-amylase, short-chain lipase or esterase, long-chain lipase, protease, cellulase, xylanase, mannanase , Spot the test strains on the screening plate, repeat 4 times for each strain on one plate, and observe the results after culturing at 30°C for 3 days.
[0075] The screening medium adopts α-amylase screening medium, esterase (short-chain lipase) screening medium, long-chain lipase screening medium, protease screening medium, cellulase screening medium, xylanase screening culture There are 7 kinds of screening media for base and mannanase, and these 7 screening media are adjusted and set on the basis of basic medium.
[0076] 1. Alpha-amylase screening medium: Add 10g of soluble starch (domestic a...
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