A method for analyzing microbial oil composition
A technology for microbial oils and bacteria, which is applied in analytical materials, material separation, measurement devices, etc., can solve the difficulty of gas chromatography analysis of triglycerides, does not mention the qualitative and quantitative analysis of monoglycerides and diglycerides, and cannot be quantitatively analyzed. Glycerides etc.
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Embodiment 1
[0014] Embodiment 1: use Mortierella alpina to prepare microbial oil
[0015] a) Preparation of spore suspension: take Mortierella alpina (preservation number: CCTCCM2013419). The bacteria were cultured for 7 days until the spores matured, and then submerged in liquid culture.
[0016] b) Seed bottle culture: Scrape the spores and mycelium on the potato dextrose agar (PDA) medium plate into an Erlenmeyer flask containing 10 ml of sterile water and 5 glass beads with a diameter of 3 mm. 200 rev / min shakers were shaken for 10 minutes, then the spore suspension was inserted into the seed bottle, the inoculum size was 10% (volume ratio), the seed bottle was placed at 25° C., and the shaker of 120 rev / min was cultivated for 36 hours. The medium used for the seed culture is: the carbon source is glucose 20g / l; the nitrogen source is: yeast powder 5g / l; pH 5.5.
[0017] c) Fermentation bottle cultivation: the bacterial concentration in the seed bottle reached 15%, was inserted into...
Embodiment 2
[0019] Embodiment 2: use Brassella trispora to prepare microbial oil
[0020] a) Strain activation: Inoculate the positive and negative strains of B. trispora into slanted PDA medium in a sterile environment, place them in an incubator, and culture them at 27°C for 5 days. After the positive and negative strains of B. trispora grow respectively, After the spores are released, scrape and wash the spores in the slant medium with sterile physiological saline under aseptic conditions, and prepare a uniform spore suspension, so that the concentrations of the positive and negative spore suspensions respectively reach: 10 3 ~10 6 spores / mL, negative bacteria 10 3 ~10 6 spores / mL;
[0021] b) Seed culture: positive and negative bacteria are respectively inoculated into the seed culture medium in the form of spore suspension, the seed culture medium is packed in a 1000mL Erlenmeyer flask, the liquid volume is 100-150mL, the culture temperature is 27°C, and the rotation speed is 220-...
Embodiment 3
[0025] Embodiment 3: use Schizochytrium to prepare microbial oil
[0026] a) Activation culture: the ampoule tube bacteria were inoculated into the mother bottle (1000ml triangular flask with 200ml culture solution), placed on a shaker (180r / min), temperature 28°C, humidity 60-70%, cultured for 48h.
[0027] b) Seed culture: insert the seed solution in the activation bottle into a 500ml Erlenmeyer flask with 100ml of medium according to the inoculum size of 5%, place it on a shaker (180r / min), temperature 28°C, humidity 60-70% , cultivated for 48h.
[0028] c) Fermentation process: After the shake flask seed liquid is connected to the first-level fermenter (1.7m3) for 2 days, it is inoculated into the second-level fermenter (12m3) for 2 days, and then it is inoculated into the third-level fermenter (45m3) for fermentation. 1:1 (v / v), the whole process adjusts the pH and dissolved oxygen and controls the foam generated during the fermentation process. The preparation method o...
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