Method for synthesizing p-hydroxybenzoic acid and esters by microvesicle bacteria
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A p-hydroxybenzoic acid, biosynthesis technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., to achieve the effects of saving energy, avoiding pollution, and reducing pollution
Inactive Publication Date: 2015-08-05
XUZHOU NORMAL UNIVERSITY
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[0009] The purpose of the present invention is to provide a method for the biosynthesis of p-hydroxybenzoic acid and p-hydroxybenzoic acid esters by bacteria of the genus Microvesicles, to solve the problem of biosynthesizing p-hydroxybenzoic acid and p-hydroxybenzoic acid esters from renewable resources
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Embodiment 1
[0080] Example 1: Biosynthesis of p-hydroxybenzoic acid:
[0081] (1) Prepare 10 ml of seawater medium containing 1% glucose;
[0082] (2) Inoculate A4B bacterial strain and cultivate for 24 hours, and the cultivation temperature is 35° C. This cultivation process is pre-cultivation, and the culture solution obtained is a pre-culture solution;
[0083] (3) prepare a 10L fermenter, and pack 8L of seawater culture medium containing 1% glucose;
[0084] (4) Inoculate 10 ml of pre-cultured bacterial liquid, cultivate at 35° C. to a stable late stage, and stop culturing after detecting that glucose is consumed by liquid chromatography;
[0085] (5) Centrifuge to remove the thallus, collect the supernatant liquid, add hydrochloric acid to adjust the pH of the liquid to acid;
[0086] (6) adding ethyl acetate equivalent to the supernatant to extract p-hydroxybenzoic acid;
[0087] (7) Collect the ethyl acetate phase containing p-hydroxybenzoic acid, and use an evaporator to evaporat...
Embodiment 2
[0088] Example 2: Biosynthesis of parabens.
[0089] (1) Prepare 10 ml of seawater medium containing 1% glucose;
[0090] (2) Inoculate A4B bacterial strain and cultivate for 24 hours, and the cultivation temperature is 35° C. This cultivation process is pre-cultivation, and the culture solution obtained is a pre-culture solution;
[0091] (3) prepare a 10L fermenter, and pack 8L of seawater culture medium containing 1% glucose;
[0092] (4) Inoculate 10 ml of pre-cultured bacterial liquid, cultivate at 35° C. to a stable late stage, and stop culturing after detecting that glucose is consumed by liquid chromatography;
[0093] (5) Centrifuge to remove the bacterium, and collect the supernatant liquid;
[0094] (6) adding ethyl acetate equivalent to the supernatant to extract p-hydroxybenzoic acid;
[0095] (7) Collect the ethyl acetate phase containing p-hydroxybenzoic acid, and use an evaporator to evaporate the ethyl acetate to obtain p-hydroxybenzoic acid esters.
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Abstract
The invention relates to a method for synthesizing p-hydroxybenzoic acid and esters by microvesicle bacteria. According to the invention, p-hydroxybenzoic acid is an organic synthetic raw material with wide usage for synthesizing liquid crystal, the esters have bacteriostatic activity and can be used as an antiseptic and widely used for preservation of food and cosmetics. The microvesicle bacteria for naturally synthesizing p-hydroxybenzoic acid and esters can be found in sea. Final accumulation concentration of p-hydroxybenzoic acid in a bacterial strain culture liquid is 10mM. The synthesized esters comprise methylparaben, ethyl ester, propyl ester, butyl ester, heptyl ester and pelargonic ester, each synthetic amount is about 2nM. For obtaining p-hydroxybenzoic acid, 2% of glucose is added in a sea culture medium for culturing A4B bacterial strain to a stable phase, hydrochloric acid is used for adjusting acidity, and ethyl acetate is used for extracting. Esters can be obtained by extracting the culture liquid under neutrallty condition. The industrial raw materials p-hydroxybenzoic acid and esters can be synthesized by microvesicle bacteria, and excessive dependence of petrifaction source by people can be reduced.
Description
technical field [0001] The invention relates to a method for synthesizing p-hydroxybenzoic acid and esters, in particular to a method for the biosynthesis of p-hydroxybenzoic acid and esters by microbubble bacteria. Background technique [0002] 4-Hydroxybenzoic acid (4HBA) is a widely used raw material for organic synthesis, mainly used in the synthesis of food, cosmetics, pharmaceutical antisepsis, antifungal agents and fungicides. In 2002, the production of 4HBA in my country was 6,000 tons, and it increased at an annual rate of 11.4%. It is estimated that the production in 2014 will reach more than 20,000 tons. One of the uses of p-hydroxybenzoic acid is the synthesis of p-hydroxybenzoic acid esters. The English name of the ester compound is Paraben, and the methyl ester is also called paraben A, and the ethyl ester is also called paraben B. Most commonly used preservatives. Another important use of p-hydroxybenzoic acid is its application in the synthesis of liquid cr...
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