阅读说明：本技术 一种d-泛解酸内酯的生产方法 (Production method of D-pantoic acid lactone ) 是由 郭恒华 张冬竹 刘磊 刘树蓬 杨林 章晖 田宋魁 于 2019-03-29 设计创作，主要内容包括：本发明属于生物合成技术领域,尤其涉及一种D-泛解酸内酯的生产方法,以DL-泛解酸内酯和含钙化合物为原料制备D-泛解酸钙,再加入酸性溶液将D-泛解酸钙酸解,即得D-泛解酸内酯溶液。采用上述方案制备的D-泛解酸内酯,所得D-泛解酸内酯光学纯度高,生产方法简单,L-泛解酸内酯仍然可以回收利用,尤其是避免了采用有机溶剂萃取中间产物而导致的高成本和高污染,适合在工业大生产中大力推广。(The invention belongs to the technical field of biosynthesis, and particularly relates to a production method of D-pantoic acid lactone. The D-pantoic acid lactone prepared by the scheme has high optical purity, the production method is simple, the L-pantoic acid lactone can still be recycled, and particularly, the high cost and high pollution caused by extracting an intermediate product by using an organic solvent are avoided, so that the method is suitable for being widely popularized in industrial mass production.)

1. A production method of D-pantoic acid lactone is characterized in that: preparing D-calcium pantoate from DL-pantoic acid lactone and calcium-containing compound, and adding acidic solution to carry out acidolysis on the D-calcium pantoate to obtain D-pantoic acid lactone solution.

2. The method for producing D-pantolactone according to claim 1, comprising the steps of:

A) adding a calcium-containing compound into the DL-pantoic acid lactone water solution, then adding D-pantoic acid lactone hydrolase, reacting for 10-20 hours at 25-35 ℃, and filtering to obtain a precipitate A;

B) and mixing the precipitate A with concentrated sulfuric acid, reacting at constant temperature, and centrifuging to remove the precipitate after the reaction is finished to obtain a solution B, namely the D-pantolactone solution.

3. The process for producing D-pantolactone according to claim 1 or 2, characterized in that: after adding a microbial liquid for producing D-pantolactone hydrolase, the enzyme activity concentration of the mixed liquid is 3-5U.

4. The process for producing D-pantolactone according to claim 1, characterized in that: adding a calcium-containing compound to a pH of 5 to 7.

5. The process for producing D-pantolactone according to claim 1, characterized in that: the calcium-containing compound is at least one of calcium carbonate, calcium oxide and calcium hydroxide.

6. The process for producing D-pantolactone according to claim 1, characterized in that: adding concentrated sulfuric acid until the pH value is 1-2, and reacting for 1-2 hours at the constant temperature of 30-80 ℃.

7. The process for producing D-pantolactone according to claim 6, characterized in that: the constant temperature condition is (50 +/-5) DEG C.

Technical Field

The invention belongs to the technical field of biosynthesis, and particularly relates to a production method of D-pantolactone.

Background

D-pantoic acid calcium is also called vitamin B5, is a calcium salt of the dextroisomer of pantothenic acid, is white powder, has no odor, bitter taste and slight moisture absorption, is used as one of B vitamins, and is an important raw material of medicines, foods, health products and the like.

D-pantothenic acid is widely present in the biological world, is a main raw material for synthesizing coenzyme A in animals, and has a main function of participating in the metabolism of sugar, fat and protein. Lack of pantothenic acid in the human body causes symptoms such as muscle soreness or spasm, adrenal insufficiency or hypofunction, inattention, numbness of hands and feet, constipation, lack of energy, exhaustion of the muscles and tendons after slight exercise, anxiety, tension, and tooth grinding. Pantothenic acid is widely applied to derivatives of medicines, feeds, foods and the like due to the unique biochemical function of the pantothenic acid. The currently more effective and widely used pantothenic acid products are calcium pantoate, panthenol, pantethine, etc.

The D-calcium pantoate is the largest one of pantothenic acid series products, is also the most main commodity form of pantothenic acid, and is widely applied to feed, medicine and food industries. D-pantoate is used as a component of coenzyme A to participate in regulating the metabolism of protein, sugar and fat, improving the color of hair and preventing diseases. Particularly, the growth and development of poultry, livestock and fish animals, and the synthesis and decomposition of fat are indispensable to the requirement of calcium pantoate. The deficiency of pantothenic acid results in growth retardation of poultry and livestock, reproductive dysfunction and adaptability reduction.

D-pantoic acid lactone (D-PL) is an important chiral intermediate for synthesizing D-pantothenic acid series products. The traditional D-pantoic acid lactone synthesis method is to hydrolyze D-pantoic acid lactone in DL-pantoic acid lactone by a microbial enzyme method to obtain L-pantoic acid lactone and D-pantoic acid, then to separate, extract and extract by utilizing ethyl acetate to obtain D-pantoic acid, and to obtain pure D-pantoic acid lactone through lactonization treatment. The method has the disadvantages that an organic reagent of ethyl acetate is needed, a large amount of organic waste liquid is generated, the production cost is increased, the environmental pollution is increased, and D-pantoic acid lactone with high optical purity can be obtained only by hydrolyzing lactone with L-configuration completely.

Disclosure of Invention

The invention aims to provide a method for producing D-pantoic acid lactone, which has higher purity of the D-pantoic acid lactone product and avoids environmental pollution caused by using organic solvents such as ethyl acetate and the like.

In order to achieve the purpose, the invention adopts the technical scheme that: preparing D-calcium pantoate from DL-pantoic acid lactone and calcium-containing compound, and adding acidic solution to carry out acidolysis on the D-calcium pantoate to obtain D-pantoic acid lactone solution.

The D-pantoic acid lactone prepared by the scheme has high optical purity, the production method is simple, the L-pantoic acid lactone can still be recycled, and particularly, the high cost and high pollution caused by extracting an intermediate product by using an organic solvent are avoided, so that the method is suitable for being widely popularized in industrial mass production.

As a preferred scheme, the production method of the D-pantoic acid lactone comprises the following specific steps:

A) adding a calcium-containing compound into the DL-pantoic acid lactone water solution, then adding D-pantoic acid lactone hydrolase, reacting for 10-20 hours at 25-35 ℃, and filtering to obtain a precipitate A;

B) and mixing the precipitate A with concentrated sulfuric acid, reacting at constant temperature, and centrifuging to remove the precipitate after the reaction is finished to obtain a solution B, namely the D-pantolactone solution.

Preferably, after the microbial liquid for producing the D-pantolactone hydrolase is added, the enzyme activity concentration of the mixed liquid is 3-5U. Wherein the enzyme activity is defined as: the production of 1. mu. mol of calcium D-pantoate per minute per ml of conversion solution is defined as 1 enzyme activity unit U. As for the method for obtaining D-pantoic lactone hydrolase, the prior art is well established, for example, the screening of D-pantoic lactone hydrolase-producing bacteria and the study of conditions for producing the enzyme are disclosed (journal of microbiology 42, No. 1, No. 2.2002, New Tang, Sunzhihao et al, pages 81-87) in a method for producing D-pantoic lactone hydrolase by culturing Fusarium moniliforme, and thus the present invention does not limit the source of D-pantoic lactone hydrolase.

Further, a calcium-containing compound is added to a pH of 5 to 7. Wherein the calcium-containing compound is at least one of calcium carbonate, calcium oxide and calcium hydroxide. In fact, the calcium-containing compound may be selected from the above calcium-containing inorganic compounds, and organic calcium-containing compounds such as calcium lactate, calcium citrate, and calcium acetate also have the effect of adjusting the pH of the mixed solution, but organic groups are introduced into the calcium-containing organic compounds, which brings trouble to subsequent product purification, and therefore inorganic calcium-containing compounds are generally used for the reaction.

Preferably, adding concentrated sulfuric acid until the pH value is 1-2, and reacting for 1-2 hours at a constant temperature of 30-80 ℃. Specifically, the constant temperature reaction temperature is (50 +/-5) DEG C.

The D-pantoic acid lactone is prepared by adopting the scheme, firstly, a calcium-containing compound and D-pantoic acid lactone hydrolase are added into DL-pantoic acid lactone, the D-pantoic acid lactone is converted into D-pantoic acid calcium precipitate, the D-pantoic acid lactone precipitate is filtered and then subjected to lactonization reaction with concentrated sulfuric acid, calcium ions and sulfate ions carried by sulfuric acid generate calcium sulfate precipitate, and the D-pantoic acid calcium lactone solution is obtained after the precipitate is filtered.

Drawings

FIG. 1 is a liquid chromatogram of the solution A obtained in step A) in example 1;

FIG. 2 is a liquid chromatogram of the solution B obtained in step B) of example 1.

Detailed Description

Firstly, preparation