Extraction and purification method of glutary-7-aminocefaphytanic acid acyltransferase
A technology of aminocephalosporanic acid acylase and purification method, applied in the direction of hydrolase, etc., can solve the problems of complex process, poor quality, serious pollution, etc., and achieve the effect of improving specific activity
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Embodiment 1
[0012] The extraction and purification process of GL-7-ACA acylase is as follows: firstly adjust the pH of the fermentation broth produced by fermentation to 7.0 with dilute hydrochloric acid, then lower the pH to 6.6 with 20% calcium chloride, and then add 2% of the bacteria chitosan, and finally adjust the pH to 7.6 with dilute alkali. Centrifuge the bacterial liquid with a high-speed centrifuge, control the activity of the bacterial liquid to 5-8u / ml, and add surfactant cetyltrimethylammonium bromide equivalent to 2% (g / l) of the bacterial liquid. Stir and soak naturally for 3 hours. During the purification process, use 20% calcium chloride solution to lower the pH of the soaked bacterial solution to 5.8, collect the enzyme serum by plate and frame filtration, and use 10% sodium hydroxide solution to increase the pH of the enzyme serum to 7.6, and then collect the serum by plate and frame filtration. The specific activity of the serum was increased from 3.4u / mg to 7.8u / mg,...
Embodiment 2
[0014] The extraction and purification process of GL-7-ACA acylase is as follows: firstly adjust the pH of the fermentation broth produced by fermentation to 7.2 with dilute hydrochloric acid, then lower the pH to 6.8 with 20% calcium chloride, and then add 2% of the bacteria chitosan, and finally adjust the pH to 7.8 with dilute alkali. Centrifuge the bacterial liquid with a high-speed centrifuge, control the activity of the bacterial liquid to 5-8u / ml, and add cetyltrimethylammonium bromide equivalent to 2% (g / l) of the bacterial liquid. Stir and soak naturally for 5 hours. During the purification process, use 20% calcium chloride solution to lower the pH of the soaked bacterial solution to 6.2, collect the enzyme serum by plate and frame filtration, and use 10% sodium hydroxide solution to increase the pH of the enzyme serum to 8.0, and then collect the serum by plate and frame filtration. The specific activity of the serum was increased from 3.4u / mg to 7.8u / mg, and the pu...
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