A strain of Rhodococcus spp. and its screening method and application
A technology of Rhodococcus luteus and Rhodococcus is applied in chemical instruments and methods, microorganism-based methods, biochemical equipment and methods, etc., to achieve the effects of low operation and maintenance cost, simple operation and simple method.
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Embodiment 1
[0026] Example 1 Isolation and purification of strains
[0027] 1. Preparation of culture medium
[0028] (1) Mineral salt medium
[0029] K 2 HPO 4 ·3H 2 O: 4.25.00g, NaH 2 PO 4 ·H 2 O: 1.00g, MgSO 4 ·7H 2 O: 0.20g, FeSO 4 ·7H 2 O: 0.012g, MnSO 4 ·H 2 O: 0.003g, ZnSO 4 ·7H 2 O: 0.003g, CoSO 4 ·7H 2 O: 0.001g, NH 4 Cl 2.0g, ultrapure water 1000mL, pH=7.3, mineral salt medium was sterilized at 121°C for 20 minutes at constant temperature.
[0030] (2) Solid enrichment medium
[0031] TryptonyeSoyaBroth (OxoidLTD, England) 30g, agar 20g, ultrapure water 1000mL, the solid enrichment medium was sterilized at 121°C for 20 minutes before use.
[0032] 2. Domestication and enrichment culture
[0033] Microbial domestication and enrichment culture is carried out in the biological activated carbon flow domestication treatment device. The water in the domestication treatment device is taken from the effluent of the biological activated carbon filter of the drinking...
Embodiment 2
[0037] Example 2 Study on the Microbiological Characteristics of the Strain of Rhodococcus spp.
[0038] 1. Colony morphology observation
[0039] Streak-inoculate the isolated and purified Rhodococcus unicellular strain into solid enrichment medium, culture at 25°C until colonies grow, and observe the colony shape. The results show that the colony characteristics of the strain are as follows: The diameter of the colonies cultured on the enrichment medium plate for 3 days is about 1-2mm, the colonies are round, raised, opaque, the center is light reddish yellow, and the edges are neat, such as figure 1 shown.
[0040] 2. Observation of cell morphology
[0041] The colony of the strain cultured on the solid enrichment medium plate for 3 days was picked, fixed with glutaraldehyde, rinsed with phosphate buffer (pH7.3), dehydrated with gradient ethanol, treated with isoamyl acetate, and dried at the critical point of carbon dioxide. Spray gold, put the sample into the observ...
Embodiment 3
[0045] Example 3 Application of Rhodococcus syringae
[0046] 1. Preparation of wet cells of Rhodococcus monocystic strain
[0047] Use an inoculation loop to take the fresh slant strains of Rhodococcus spp., streak and inoculate it on the solid enrichment medium, and culture it at 25°C for 3 days to obtain a medium plate with obvious colonies, and obtain fresh stalks Rhodococcus wet cells.
[0048] 2. Removal of 5 kinds of nitrosamine substances
[0049] Pick the wet cells of the strain Rhodococcus syringae and inoculate them in mineral salt medium, and cultivate them at 25°C under dark conditions, wherein, inoculate 0.6g of wet cells per 100mL of mineral salt medium; mineral salt culture The mass concentrations of the five nitrosamines in the base were 200ng / L. The detection method was based on the previous research and was determined by ultra-high liquid chromatography tandem mass spectrometry (UPLC / MS / MS). 1 day, 3 days, 5 days, 7 days, the mass concentration value of...
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