阅读说明：本技术 一种提高全细胞催化制备糠酸的方法 (Method for improving catalytic preparation of furoic acid by whole cells ) 是由 徐勇 杜根来 周鑫 于 2019-11-22 设计创作，主要内容包括：本发明公开了一种提高全细胞催化制备糠酸的方法,其中,一种提高全细胞催化制备糠酸的方法,其包括,将糠醛液、能源物质、菌体通风搅拌；调节pH和温度,进行控温反应；加入所述能源物质后搅拌,即可；一种连续化糠酸生产工艺,其中：包括,将糠醛液、能源物质、菌体通风搅拌；调节pH和温度,进行控温反应；加入所述能源物质后搅拌；分离所述氧化葡萄糖酸杆菌,重复前述步骤。该方法获得的全细胞催化制备糠酸液,反应6h产品糠酸最高浓度即可达40g/L以上,糠酸单位产率超过6.5g/L/h,24h后糠酸累积浓度可达120g/L以上。(The invention discloses a method for improving the catalytic preparation of furoic acid by whole cells, wherein the method for improving the catalytic preparation of furoic acid by whole cells comprises the following steps of ventilating and stirring furfural liquid, energy substances and thalli; adjusting the pH and the temperature, and carrying out temperature control reaction; adding the energy substance and then stirring; a continuous furoic acid production process, wherein: comprises ventilating and stirring furfural liquid, energy substances and thalli; adjusting the pH and the temperature, and carrying out temperature control reaction; adding the energy substance and stirring; and (3) separating the gluconobacter oxydans, and repeating the steps. The total cell catalytic prepared furoic acid solution obtained by the method has the advantages that the highest concentration of furoic acid products can reach more than 40g/L after 6 hours of reaction, the unit yield of furoic acid exceeds 6.5g/L/h, and the cumulative concentration of furoic acid can reach more than 120g/L after 24 hours.)

1. A method for improving the catalytic preparation of furoic acid by whole cells is characterized in that: comprises the steps of (a) preparing a mixture of a plurality of raw materials,

ventilating and stirring furfural liquid, energy substances and thalli;

adjusting the pH and the temperature, and carrying out temperature control reaction;

and adding the energy substance and stirring.

2. The method for enhancing whole-cell catalytic production of furoic acid according to claim 1, wherein: the furfural liquid is an arbitrary combined reaction substrate containing furfural or comprising furfural; the energy source substance comprises one or more carbon sources of saccharides, lipids, alcohols and amino acids; the thallus comprises gluconobacter oxydans and all genetic mutation or gene recombination strains taking the gluconobacter oxydans as starting strains; wherein the cell concentration ratio of the furfural liquid, the energy substance and the thallus is 1 (0.1-0.5) to 0.03-0.3.

3. The method for enhancing whole-cell catalytic production of furoic acid according to claim 1, wherein: the energy substance is added and then stirred, and the operation mode of semi-continuous or continuous feeding is adopted.

4. A method for improving the whole-cell catalytic production of furoic acid according to any one of claims 1 to 3, wherein: the aeration stirring is carried out at a stirring speed of 300-500 r/min and an oxygen introduction amount of 0.1-3 vvm, the pH and the temperature are adjusted to 5.0-6.5, and the temperature is adjusted to 25-35 ℃.

5. A method for improving the whole-cell catalytic production of furoic acid according to any one of claims 1 to 3, wherein: the thallus is gluconobacter oxydans, and the energy substance is one or more of glucose, sorbitol, glycerol and mannitol.

6. A continuous furoic acid production process is characterized in that: comprises the steps of (a) preparing a mixture of a plurality of raw materials,

ventilating and stirring furfural liquid, energy substances and thalli;

adjusting the pH and the temperature, and carrying out temperature control reaction;

adding the energy substance and stirring;

and (3) separating the gluconobacter oxydans, and repeating the steps.

7. The continuous furoic acid production process of claim 6, wherein: the furfural liquid is an arbitrary combined reaction substrate containing furfural or comprising furfural; the energy source substance comprises one or more carbon sources of saccharides, lipids, alcohols and amino acids; the thallus comprises gluconobacter oxydans and all genetic mutation or gene recombination strains taking the gluconobacter oxydans as starting strains;

wherein the cell concentration ratio of the furfural liquid, the energy substance and the thallus is 1 (0.1-0.5) to 0.03-0.3.

8. The method for enhancing whole-cell catalytic production of furoic acid according to claim 7, wherein: the thallus is gluconobacter oxydans, and the energy substance is one or more of glucose, sorbitol, glycerol and mannitol.

9. The method for improving whole-cell catalytic production of furoic acid according to any one of claims 6 to 8, wherein: after the energy substances are added, stirring is carried out, wherein a semi-continuous or continuous feeding operation mode is adopted, and the separation is centrifugal separation; the aeration stirring is performed at a stirring speed of 200-500 r/min and an oxygen introduction amount of 0.1-3 vvm, and the pH and the temperature are adjusted to 5.0-6.5 and 25-35 ℃.

10. A whole-cell catalytic preparation of a furoic acid solution is characterized in that: the furfural liquid is obtained after the fermentation of thalli for 24 hours, and the cumulative concentration of the furoic acid can reach more than 120 g/L.

Technical Field

The invention belongs to the technical field of biological engineering and chemical engineering, and particularly relates to a method for improving the catalytic preparation of furoic acid by whole cells.

Background

Furoic acid, also known as α -furancarboxylic acid, 2-furancarboxylic acid or furancarboxylic acid, is an important organic synthetic raw material, which can be converted to produce derivatives such as ester, acyl chloride, anhydride, amide, etc., furoic acid and its derivatives are important raw materials for synthesizing α -furancarboxylic acid ester perfume, drugs, pesticides, plasticizers, and tetrahydrofurfuryl acid obtained by catalytic hydrogenation of furoic acid is a very widely used medical intermediate, which can be used for preparing cephalosporin antibiotic drug intermediates and chiral auxiliary agents, drugs for treating hypertension of oxazolines and drug synthesis for treating prostate cancer, etc.

Disclosure of Invention

This section is for the purpose of summarizing some aspects of embodiments of the invention and to briefly introduce some preferred embodiments. In this section, as well as in the abstract and the title of the invention of this application, simplifications or omissions may be made to avoid obscuring the purpose of the section, the abstract and the title, and such simplifications or omissions are not intended to limit the scope of the invention.

The present invention has been made in view of the above-mentioned technical drawbacks.

Therefore, as one aspect of the invention, the invention overcomes the problems of low yield of the product furoic acid and low utilization rate of thalli in the prior art; and provides a method for improving the whole-cell catalytic preparation of furoic acid aiming at the problem of product inhibition caused by accumulation of product furoic acid.

In order to solve the technical problems, the invention provides the following technical scheme: a method for improving the catalytic preparation of furoic acid by whole cells comprises the steps of ventilating and stirring furfural liquid, energy substances and thalli; adjusting the pH and the temperature, and carrying out temperature control reaction; and adding the energy substance and stirring.

As a preferred scheme of the method for improving the whole-cell catalytic preparation of furoic acid, the method comprises the following steps: the furfural liquid is an arbitrary combined reaction substrate containing furfural or comprising furfural; the energy source substance comprises one or more carbon sources of saccharides, lipids, alcohols and amino acids; the thallus comprises gluconobacter oxydans and all genetic mutation or gene recombination strains taking the gluconobacter oxydans as starting strains; wherein the cell concentration ratio of the furfural liquid, the energy substance and the thallus is 1 (0.1-0.5) to 0.03-0.3.

As a preferred scheme of the method for improving the whole-cell catalytic preparation of furoic acid, the method comprises the following steps: the energy substance is added and then stirred, and the operation mode of semi-continuous or continuous feeding is adopted.

As a preferred scheme of the method for improving the whole-cell catalytic preparation of furoic acid, the method comprises the following steps: the aeration stirring is carried out at a stirring speed of 300-500 r/min and an oxygen introduction amount of 0.1-3 vvm, the pH and the temperature are adjusted to 5.0-6.5, and the temperature is adjusted to 25-35 ℃.

As a preferred scheme of the method for improving the whole-cell catalytic preparation of furoic acid, the method comprises the following steps: the thallus is gluconobacter oxydans, and the energy substance is one or more of glucose, sorbitol, glycerol and mannitol.

As one aspect of the present invention, the present invention provides a continuous furoic acid production process, wherein: comprises ventilating and stirring furfural liquid, energy substances and thalli; adjusting the pH and the temperature, and carrying out temperature control reaction; adding the energy substance and stirring; and (3) separating the gluconobacter oxydans, and repeating the steps.

The preferable scheme of the continuous furoic acid production process of the invention is as follows: the furfural liquid is an arbitrary combined reaction substrate containing furfural or comprising furfural; the energy source substance comprises one or more carbon sources of saccharides, lipids, alcohols and amino acids; the thallus comprises gluconobacter oxydans and all genetic mutation or gene recombination strains taking the gluconobacter oxydans as starting strains; wherein the cell concentration ratio of the furfural liquid, the energy substance and the thallus is 1 (0.1-0.5) to 0.03-0.3.

The preferable scheme of the continuous furoic acid production process of the invention is as follows: the thallus is gluconobacter oxydans, and the energy substance is one or more of glucose, sorbitol, glycerol and mannitol.

The preferable scheme of the continuous furoic acid production process of the invention is as follows: after the energy substances are added, stirring is carried out, wherein a semi-continuous or continuous feeding operation mode is adopted, and the separation is centrifugal separation; the aeration stirring is performed at a stirring speed of 200-500 r/min and an oxygen introduction amount of 0.1-3 vvm, and the pH and the temperature are adjusted to 5.0-6.5 and 25-35 ℃.

As one aspect of the present invention, the present invention provides a whole-cell catalytic preparation of a furoic acid solution, which comprises: the furfural liquid is obtained after the fermentation of thalli for 24 hours, and the cumulative concentration of the furoic acid can reach more than 120 g/L.

The invention has the beneficial effects that:

compared with a chemical method, the method has the advantages that the substrate conversion rate is high, the purity of the furoic acid product is high, and the wastewater treatment is simple; under the action of an auxiliary carbon source, the yield and the yield of furfural are effectively improved, the highest concentration of the furoic acid product after 6 hours of reaction can reach more than 40g/L, and the unit yield of the furoic acid exceeds 6.5 g/L/h; in addition, the catalytic somatic cells can be recycled for multiple times to carry out a new cycle of furoic acid preparation reaction, and the cumulative concentration of furoic acid can exceed 120g/L through three times of cell recycling and re-catalysis within 24 hours; the method has good practicability and industrialization prospect.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without inventive exercise. Wherein:

FIG. 1 is a diagram showing the reaction scheme of furfural production by whole-cell catalytic oxidation in example 1;

FIG. 2 is a reaction scheme of furfural preparation by whole-cell catalytic oxidation in example 2;

FIG. 3 is a reaction scheme of furfural preparation by whole-cell catalytic oxidation in example 4;

FIG. 4 is a reaction scheme of furfural production by whole-cell catalytic oxidation in example 10;

FIG. 5 is a reaction scheme of furfural production by whole-cell catalytic oxidation in example 11.

Detailed Description

In order to make the aforementioned objects, features and advantages of the present invention comprehensible, embodiments accompanied with examples are described in detail below.

In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention, but the present invention may be practiced in other ways than those specifically described and will be readily apparent to those of ordinary skill in the art without departing from the spirit of the present invention, and therefore the present invention is not limited to the specific embodiments disclosed below.

Furthermore, reference herein to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one implementation of the invention. The appearances of the phrase "in one embodiment" in various places in the specification are not necessarily all referring to the same embodiment, nor are separate or alternative embodiments mutually exclusive of other embodiments.