Method for extracting concealed acidophilic bacteria cohesive action-beta-hydroxy-butanoic acid ester
A technology of hydroxybutyrate and extraction method, which is applied in the field of extracting intracellular poly-β-hydroxybutyrate of hidden acidophilus, which can solve the serious degradation of PHB molecules and other problems
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[0008] 1. Culture and collection of strains
[0009] Acidiphilium cryptum strain DX1-1 was cultured to the mid-logarithmic growth phase, and the bacterial cells were collected by centrifugation at 10,000 r / min for 10 min, and the bacterial cells were washed twice with sterile water (pH 3.0). Resuspend the precipitated sludge with 20 mL of sterile water.
[0010] 2. Ultrasonic crushing
[0011] Put the centrifuge tube containing the bacterial solution in a beaker with ice cubes in advance, and ultrasonically break it according to the following procedure: work for 2s, interval of 4s, work 10-30 times.
[0012] 3. Dispersed NaClO / Chloroform Extraction of PHB
[0013] Add 80 mL of sodium hypochlorite solution (pH10) / chloroform with a volume ratio of 2:1 to 1:2 to the liquid after ultrasonic crushing, shake it on a shaker at 30°C for 60 minutes, and centrifuge at 8000 r / m to divide it into three layers of sodium chlorate solution. The middle layer is not broken. Parietal cells a...
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