Schizochytrium sp.TIO1101 strain with high-yield DHA (docosahexaenoic acid) and fermentation method thereof
A technology of Schizochytrium algae and fermentation medium, which is applied to high DHA-producing Schizochytrium algae strains and the field of fermentation thereof, can solve the problem of low yield and the like, and achieve the effects of high DHA content and simple ingredients
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[0023] Preparation method of trace element compound solution: Na 2 · EDTA 6.0g, FeCl 3 ·6H 2 O 0.29g, H 3 BO 3 6.84g, MnCl 2 4H 2 O 0.86g, ZnCl 2 0.06g, CoCl 2 6H2O 0.026g, NiSO 4 ·6H 2 O 0.052g, CuSO 4 ·5H 2 O0.002g, NaMoO 4 2H 2 O 0.005g, make up 1L with distilled water.
[0024] The preparation method of vitamin complex solution: vitamin B1 (thiamin) 100mg, vitamin H (biotin) 0.5mg, vitamin B12 (cyanocobalamin) 0.5mg, distilled water to supplement 1L.
[0025] The preparation method of the fermentation medium: 120g of glucose, 20g of yeast extract, 15g of peptone, 3g of potassium dihydrogen phosphate, 2g of magnesium sulfate, 10ml of trace element compound solution, 5ml of vitamin compound solution, and 0.5 × seawater (diluted with fresh water 1 times seawater) to 1L.
Embodiment 1
[0026] Example 1, the isolation and identification of Schizochytrium TIO1101
[0027] 1. Isolation of Schizochytrium TIO1101 (pine pollen fishing method)
[0028] Take the decayed fallen leaves from the mangrove forest in Yundang Lake, Xiamen, cut them into small discs with a diameter of about 1.5 cm, wash them with sterile seawater containing 1 g / L of streptomycin and penicillin, and inoculate them on solid separation plates. Add about 3mL of sterile seawater and a small amount of sterile pine pollen to the plate, culture 4-5d at 28°C, pick pine pollen, and observe the culture that adheres to and grows on the pine pollen. The pine pollen of the adherent culture was then transferred to a new plate for streak separation until a pure culture was obtained. The resulting algae was named TIO1101.
[0029] 2. Identification of Schizochytrium TIO1101
[0030] 1. Observation of algae colony and cell morphology
[0031]After the culture was cultured on the isolation and purificatio...
Embodiment 2
[0045] Embodiment 2, the high-density heterotrophic fermentation of Schizochytrium TIO1101
[0046] 1. Fermentation
[0047] Add 6L of fermentation medium in a 10L fermenter, and then add 0.6L of Schizochytrium TIO1101 bacterial liquid, so that the concentration of Schizochytrium TIO1101 in the medium is about 1.0×10 6 Individual / mL; fermentation culture conditions are: temperature 25°C, pH value 6.0, rotation speed 300rpm (200-400rpm is acceptable), dissolved oxygen 10%, culture for 96h; carry out 10L fermentor fed-batch culture: from the moment of fermentation 12h Start to add glucose solution (composed of glucose and water, the concentration of glucose is 50g / L) and yeast extract solution (made up of yeast extract and water, the concentration of yeast extract is 10g / L) until the moment of fermentation 50h, the glucose solution The flow rate is 50mL / h, and the flow rate of the yeast extract solution is 20mL / h.
[0048] 2. Detection of biomass, oil and DHA content
[0049]...
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