Method for producing palmitoleic acid from Eustigmatophyceae microalgae
A technology of euphratica and palmitoleic acid, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of high cost, unsuitable for large-scale production, and low purity of palmitoleic acid, and achieve The effect of simple method
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Embodiment 1
[0039] Example 1 The optimal initial nitrogen concentration of E.cf.polyphem highly accumulating palmitoleic acid
[0040] Experimental material: E.cf.polyphem (E.cf.polyphem) (expanded and cultivated)
[0041] Medium preparation: mBG-11 liquid medium, 1.5g / L NaNO 3 , 0.04g / L K 2 HPO 4 , 0.075g / LMgSO 4 ·7H 2 O, 0.036g / L CaCl 2 2H 2 O, 0.006g / L citric acid, 0.006g / L FeCl 3 ·6H 2 O, 0.001g / LEDTA, 0.02g / L Na 2 CO 3 , 2.860g / L H 3 BO 3 , 1.810g / L MnCl 2 4H 2 O, 0.391g / L Na 2 MoO 4 2H 2 O, 0.079g / L CuSO 4 ·5H 2 O, 0.220g / L ZnSO 4 ·7H 2 O, 1L deionized water, sterilized at 121°C for 20min. where NaNO 3 As a nitrogen source, 4 different initial concentrations were set, namely 18mM (1.5g), 9mM (0.75g), 3mM (0.25g) and 1mM (0.08g).
[0042] experiment method:
[0043] The strains of E.cf.polyphem that had been expanded were divided according to the OD 750 = 0.60 ± 0.02 initial inoculation density into the optical columnar glass bioreactor (diameter × height = 3...
Embodiment 2
[0047] Phase changes in the growth and palmitoleic acid accumulation of E.cf.polyphem (E.cf.polyphem) under nitrogen stress conditions (1mM) in Example 2
[0048] Experimental material: Euophthalmos bordetii-like (expansion and cultivation have been carried out)
[0049] Medium preparation: mBG-11 liquid medium, 0.08g / L NaNO 3 , 0.04g / L K 2 HPO 4 , 0.075g / LMgSO 4 ·7H 2 O, 0.036g / L CaCl 2 2H 2 O, 0.006g / L citric acid, 0.006g / L FeCl 3 ·6H 2 O, 0.001g / LEDTA, 0.02g / L Na 2 CO 3 , 2.860g / L H 3 BO 3 , 1.810g / L MnCl 2 4H 2 O, 0.391g / L Na 2 MoO 4 2H 2 O, 0.079g / L CuSO 4 ·5H 2 O, 0.220g / L ZnSO 4 ·7H 2 O, 1L deionized water, sterilized at 121°C for 20min.
[0050] experiment method:
[0051] The euopteris strains that have been expanded were divided according to the OD 750=0.60±0.02 initial inoculation density was inserted into mBG-11 liquid medium with an initial nitrogen concentration of 1 mM, and cultured in a bright columnar glass bioreactor with diameter×heigh...
Embodiment 3
[0053] The preparation of palmitoleic acid in the embodiment 3 classes E.cf.polyphem (E.cf.polyphem)
[0054] Experimental material: Freeze-dried algae powder of E.cf.polyphem under the culture condition of 1mM nitrogen concentration
[0055] experiment method:
[0056] (1) Oil extraction: take freeze-dried algae powder and 95% ethanol and mix according to the ratio of m / v=1:30, put it in an ultrasonic cleaning machine and continue ultrasonic treatment for 2.5 hours. After ultrasonic pretreatment, the flask containing the algae solution was placed in a constant temperature water bath at 50° C., and extracted for 4 hours with continuous stirring (200 rpm) by an electric stirrer. After the extraction, centrifuge at 3500rpm for 5min, collect the supernatant, and continue to repeat the above extraction steps for 3 to 4 times for algae. After combining the supernatants, use a rotary evaporator (50° C.) to recover ethanol to obtain total lipids. Detected by gas chromatography, as...
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