阅读说明：本技术 大环内酯类抗生素发酵调控的方法 (Method for regulating fermentation of macrolide antibiotics ) 是由 王欣荣 张雪霞 褚以文 郑智慧 张新宜 路新华 赵克雷 高健 林家富 黄挺 任凤 于 2020-03-02 设计创作，主要内容包括：本发明为大环内酯类抗生素发酵调控的方法。解决传统方法发酵过程不容易控制,耗能高,目标产物产量低的问题。包括以下步骤：在大环内酯类抗生素产生菌初级代谢阶段,培养温度按正常要求控制在27.0-30.0℃,菌丝快速生长；在大环内酯抗生素产生菌完成对数生长期后降低发酵培养温度到18.0-27.0℃,促进目标产物大量合成,所述大环内酯抗生素为采用PKS生物合成途径发酵产生的抗生素及其同系物。本发明工艺简单易行,易于操作,非常适合商业化生产。(The invention relates to a method for regulating and controlling fermentation of macrolide antibiotics. Solves the problems of difficult control, high energy consumption and low yield of target products in the traditional method. The method comprises the following steps: in the primary metabolism stage of macrolide antibiotic producing bacteria, the culture temperature is controlled to be 27.0-30.0 ℃ according to normal requirements, and hyphae grow rapidly; reducing the fermentation culture temperature to 18.0-27.0 ℃ after the log growth period of macrolide antibiotic producing bacteria is completed, and promoting the mass synthesis of a target product, wherein the macrolide antibiotic is produced by fermentation by adopting a PKS biosynthesis pathway and a homologue thereof. The method has the advantages of simple and easy process, easy operation and suitability for commercial production.)

1. A method for regulating fermentation of macrolide antibiotics is characterized by comprising the following steps:

in the primary metabolism stage of macrolide antibiotic producing bacteria, the culture temperature is controlled to be 27.0-30.0 ℃ according to normal requirements, and hyphae grow rapidly; reducing the fermentation culture temperature to 18.0-27.0 ℃ after the growth period of the bacteria of the macrolide antibiotic producing bacteria is finished, and promoting the mass synthesis of the target product, wherein the macrolide antibiotic is the antibiotic produced by fermentation by adopting a PKS biosynthesis pathway and a homologue thereof.

2. The method for fermentation control of macrolide antibiotics according to claim 1, wherein the period of time for lowering the fermentation culture temperature after completion of the growth of the macrolide antibiotic-producing strain is at least two, and the fermentation culture temperature in each period of time is different, and is higher and lower.

3. The method for fermentation regulation of macrolide antibiotics according to claim 1, characterized in that, the macrolide antibiotics comprise sirolimus, tacrolimus, avermectin, spinosad and leucomycin.

4. The method for controlling fermentation of a macrolide antibiotic according to claim 1, wherein the macrolide antibiotic-producing bacterium completes a growth period of a strain 15 to 96 hours after the start of fermentation.

5. The method for fermentation control of macrolide antibiotics according to claims 1 to 4, wherein the fermentation culture temperature is lowered from 27.0-30.0 ℃ to 22.0-26.0 ℃ after 15-96 hours after the growth phase of the macrolide antibiotic-producing strain is completed, thereby promoting the mass synthesis of the target product.

6. The method for fermentation control of macrolide antibiotics according to claims 1 to 4, wherein the fermentation culture temperature is decreased from 27.0-30.0 ℃ to 23.0-25.0 ℃ after 15-96 hours from the completion of the growth phase of the bacterial strain of macrolide antibiotic-producing bacteria, thereby promoting the mass synthesis of the target product.

7. A method for fermentation regulation of macrolide antibiotics according to claims 1-3, characterized in that the whole fermentation period is prolonged 1-4 days compared to the conventional fermentation period.

8. A method for regulating fermentation of macrolide antibiotics is characterized by comprising the following steps:

in the primary metabolism stage of macrolide antibiotic producing bacteria, the culture temperature is controlled according to the traditional requirement, and hyphae grow rapidly; after 15-96 hours of logarithmic growth phase completion of macrolide antibiotic producing bacteria, the large-scale synthesis of target products is promoted by reducing the fermentation culture temperature by 3-8 ℃, wherein the macrolide antibiotic is produced by fermentation by adopting a PKS biosynthesis pathway and homologs thereof.

9. Method for the fermentation regulation of macrolide antibiotics according to claim 7, characterized in that the macrolide antibiotic is erythromycin.

Technical Field

The invention relates to the technical field of microbial medicines, in particular to a macrolide antibiotic fermentation regulation and control method based on culture temperature regulation and control.

Background

Macrolide antibiotics such as Sirolimus (Sirolimus), Tacrolimus (Tacrolimus), Doramectin (Doramectin), Spinosyn (Spinosyn) and leucomycin (Kitasamycin) belong to polyketides in structure, and the biosynthesis process of the polyketide structure also shows similar rules. Polyketide synthase (PKS) is a multienzyme complex that catalyzes the formation of polyketides, catalyzing the synthesis of the basic backbone of macrolide antibiotics with a long carbon chain structure, which is polymerized from lower fatty acids. The starter units for polyketide synthesis and the short chain fatty acid units required for carbon chain extension, such as acetic acid and propionic acid, have been accumulated during the primary metabolic stages of the microorganism. In the middle and later period of fermentation, the higher fermentation temperature is continuously maintained, so that the microorganisms can quickly consume raw materials such as sugar, grease or amino acid which generate short-chain fatty acid units to form waste and lose the waste, and the requirement of product synthesis cannot be continuously met. Meanwhile, a large amount of organic acid can be generated due to high carbon source consumption, so that the pH value and dissolved oxygen of the fermentation liquor can be reduced rapidly, and the pH value can be increased rapidly after the carbon source is consumed completely, so that the fermentation process is not easy to control.

Disclosure of Invention

The invention aims to provide a method for regulating and controlling fermentation of macrolide antibiotics, which has the advantages of simple process, easy control, low energy consumption, improved equipment utilization rate, reduced consumption of nutrient components and obviously improved yield of target products.

The invention is realized by the following steps:

a method for fermentation regulation of macrolide antibiotics, comprising the steps of:

in the primary metabolism stage of macrolide antibiotic producing bacteria, the culture temperature is controlled to be 27.0-30.0 ℃ according to normal requirements, and hyphae grow rapidly; after the growth period of the bacteria of the macrolide antibiotic producing bacteria is finished, the large-scale synthesis of a target product is promoted by reducing the fermentation culture temperature to 18.0-27.0 ℃, wherein the macrolide antibiotic is the antibiotic and the homologue thereof produced by fermentation by adopting a PKS biosynthesis pathway.

The time periods for reducing the fermentation culture temperature after the macrolide antibiotic producing bacteria complete the thallus growth period are at least two, wherein the fermentation culture temperature in each time period is different and is higher and lower.

The macrolide antibiotics include Sirolimus (Sirolimus), Tacrolimus (Tacrolimus), Avermectin (Avermectin), Spinosyn (Spinosyn), and leucomycin (Kitasamycin).

The growth period of the macrolide antibiotic producing bacteria after completing the thallus is 15-96 hours after the beginning of the fermentation.

After the macrolide antibiotic producing strain finishes the growth period of thalli, namely 15 to 96 hours, the fermentation culture temperature is reduced from 27.0 to 30.0 ℃ to 22.0 to 26.0 ℃, and the mass synthesis of a target product is promoted.

After 15-96 hours of the macrolide antibiotic producing bacteria completing the logarithmic growth phase, the fermentation culture temperature is reduced from 27.0-30.0 ℃ to 23.0-25.0 ℃, and the mass synthesis of the target product is promoted.

The whole fermentation period is prolonged by 1-4 days compared with the traditional fermentation period.

A method for fermentation regulation of biosynthesis of macrolide antibiotics, comprising the steps of:

in the primary metabolism stage of macrolide antibiotic producing bacteria, the culture temperature is controlled according to the traditional requirement, and hyphae grow rapidly; after 15-96 hours of the growth period of the bacteria is finished, the macrolide antibiotic producing bacteria promote the mass synthesis of the target product by reducing the fermentation culture temperature by 3-8 ℃, wherein the macrolide antibiotic is the antibiotic produced by fermentation by adopting a PKS biosynthesis pathway and a homologue thereof.

The macrolide antibiotic is Erythromycin (Erythromycin).

The invention has the following advantages:

the invention adopts two-section temperature control strategy, the primary metabolism stage of macrolide antibiotic producing bacteria, in the primary metabolism stage of macrolide antibiotic producing bacteria, the culture temperature is controlled according to normal requirements, and hyphae grow rapidly; after the macrolide antibiotic producing strain finishes the growth period of thalli, the large-scale synthesis of a target product is promoted by obviously reducing the fermentation culture temperature.

According to the invention, the fermentation culture temperature is obviously reduced after the macrolide antibiotic producing strain finishes logarithmic growth phase, the pH value and dissolved oxygen change of fermentation liquor in the fermentation process are found to be stable, the control is easy, and the yield of a target product is obviously improved. By implementing the cooling process, the utilization rate of equipment is improved, the consumption of nutrient components is reduced, and energy conservation and emission reduction are realized. The method has simple and easy process and easy operation, and is very suitable for commercial production.

Detailed Description

The following examples are given to further illustrate the embodiments of the present invention. The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention. The invention covers all possible alternatives, modifications and equivalents within the scope of the claims. The specific techniques or conditions not mentioned in the following examples are all the conventional techniques or conditions, or the techniques or conditions described in the literature in the field, or the product specification.