Pseudomonas fluorescens and application thereof in biosynthesizing methionine
A Pseudomonas fluorescens and substrate technology, applied in the direction of microorganism-based methods, microorganisms, biochemical equipment and methods, etc., can solve the problem of increasing investment in equipment, increasing cost recovery and processing salt, not meeting green chemistry and atomic economy, etc. problems, to achieve the effect of a wide range of application prospects
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Embodiment 1
[0037] Embodiment 1: the screening of new bacterial strain ZJB-12001
[0038]Inoculate the enrichment medium with the soil samples collected from Zhejiang Province and sewage samples from pharmaceutical factories; after 72 hours of enrichment culture on a shaker at 30°C and 150rpm, take 2ml of the culture solution and inoculate it on the fresh enrichment medium. Secondary culture, so repeated 3 cycles. After the last dilution of the culture solution, spread it on the beef extract peptone plate medium, culture at 30°C for 24-72 hours to obtain a single colony, pick a single colony and inoculate it on the slant medium for preservation. Pick an inoculation loop strain from the slant culture medium and inoculate it into the fermentation medium, shake and culture at 30°C and initial pH 7.0-7.5 for 48 hours, centrifuge, discard the supernatant, and take the precipitate to obtain the enzyme-containing wet bacteria ; The enzyme-containing wet bacteria were suspended in the pH7.0 phos...
Embodiment 2
[0041] Embodiment 2: Identification of new bacterial strain ZJB-09150
[0042] In Example 1, a bacterial strain with the strongest activity was obtained from the soil through enrichment culture and then diluted coating method through high-performance liquid chromatography detection and screening. The label is ZJB-12001. figure 1 As shown, the diameter of the colony is about 1.0-2.0 cm, and the colony is milky white, spherical, bright, moist, with neat edges and slightly raised surface. Electron microscope scanning photo see figure 2 As shown, there are 3-6 polar flagella.
[0043] Using the Biolog (GENⅢ) automatic microbial identification system, 94 phenotypic tests were carried out on strain ZJB-12001, including 71 carbon source utilization tests and 23 chemical sensitivity tests (Biolog microbial automatic identification reagents and culture media were purchased from Biolog Company), the metabolic fingerprint was analyzed by Biolog reader, and the identification results o...
Embodiment 3
[0051] Example 3: Preparation of biocatalyst Pseudomonas fluorescens ZJB-12001 enzyme-containing wet thallus
[0052] (1) Incline cultivation:
[0053] The Pseudomonas fluorescens ZJB-12001 (CCTCC No: M2012449) obtained in Example 2 was inoculated into the slant medium, cultured at 30°C for 48 hours, and the slant bacteria were obtained; the final concentration of the slant medium was composed of: peptone 10.0g / L, beef extract 5.0g / L, sodium chloride 5.0g / L, agar 20.0g / L, pH7.2.
[0054] (2) Seed cultivation:
[0055] Pick a ring of bacteria from the slant of step (1), inoculate it into 50.0ml of sterile seed medium, and cultivate it at 30°C for 24 hours to obtain the seed liquid; the final concentration of the seed medium is composed of: glucose 10.0g / L, Sodium glutamate 10.0g / L, yeast powder 3.0g / L, KH 2 PO 4 0.75g / L, K 2 HPO 4 ·3H 2 O0.75g / L, MgSO 4 ·7H 2 O0.2g / L, caprolactam 1.0g / L, pH7.0, solvent is water. The medium was sterilized at 121°C for 20 minutes.
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