阅读说明：本技术 一种赤霉酸GA3≧7.5g/L的发酵方法 (Fermentation method of gibberellic acid GA3 ≧ 7.5g/L ) 是由 景飞江 王敏 熊仁科 吴红波 张俊 左建英 徐旭 杨怀亮 李玉婷 于 2021-09-30 设计创作，主要内容包括：本申请公开了一种赤霉酸GA3≧7.5g/L的发酵方法,该发酵方法的空气流量为变速空气流量,该发酵方法的发酵培养基包括：玉米蛋白粉1～9重量份；磷酸二氢钾1～10重量份；小分子有机碳源1～30重量份；植物油0.5～5重量份；硫酸镁0.5～2重量份；硫酸铵0.5～2重量份；微量元素0.1～1重量份；大豆蛋白1～10重量份；麦麸1～9重量份；以及吐温0.001-0.5重量份。本申请发酵方法发酵出中霉酸含量≧7.5g/L。(The application discloses a fermentation method of gibberellic acid GA3 ≧ 7.5g/L, the air flow of the fermentation method is variable speed air flow, and a fermentation medium of the fermentation method comprises: 1-9 parts of corn protein powder; 1-10 parts by weight of monopotassium phosphate; 1-30 parts by weight of a small-molecular organic carbon source; 0.5-5 parts by weight of vegetable oil; 0.5-2 parts by weight of magnesium sulfate; 0.5-2 parts by weight of ammonium sulfate; 0.1-1 part by weight of trace elements; 1-10 parts by weight of soybean protein; 1-9 parts of wheat bran; and 0.001-0.5 part by weight of tween. The content of the mycolic acid in the fermentation by the fermentation method is not less than 7.5 g/L.)

1. A fermentation method of gibberellic acid GA3 ≧ 7.5g/L, this method transfers the seed liquid of gibberellic acid into the fermentation cylinder and ferments, characterized by: the air flow rate of the fermentation method is variable speed air flow rate, and the fermentation culture medium of the fermentation method comprises:

1-9 parts of corn protein powder;

1-10 parts by weight of monopotassium phosphate;

1-30 parts by weight of a small-molecular organic carbon source;

0.5-5 parts by weight of vegetable oil;

0.5-2 parts by weight of magnesium sulfate;

0.5-2 parts by weight of ammonium sulfate;

0.1-1 part by weight of trace elements;

1-10 parts by weight of soybean protein;

1-9 parts of wheat bran; and

0.001-0.5 weight portion of tween.

2. The fermentation method of gibberellic acid GA3 ≧ 7.5g/L according to claim 1, wherein the fermentation temperature is 28-30 ℃, the fermentation time is 8-10 days, and the fermentation pressure is 0.01-0.05 MPa.

3. The fermentation method of gibberellic acid GA3 ≧ 7.5g/L according to any one of claims 1-2, wherein the small-molecule organic carbon source is sucrose or glucose.

4. The fermentation method of gibberellic acid GA3 ≧ 7.5g/L according to any one of claims 1-2, wherein the small-molecule organic carbon source is sucrose and glucose.

5. The fermentation method of gibberellic acid GA3 ≧ 7.5g/L according to claim 5, wherein the sucrose: the weight ratio of glucose is 1-2: 2 to 1.

6. The fermentation method of gibberellic acid GA3 ≧ 7.5g/L according to any one of 1-5, wherein the microelements are ferrous sulfate, zinc sulfate, manganese sulfate, sodium molybdate, copper sulfate and cobalt chloride.

7. The fermentation method of gibberellic acid GA3 ≧ 7.5g/L is characterized in that the weight ratio of ferrous sulfate, zinc sulfate, manganese sulfate, sodium molybdate, copper sulfate and cobalt chloride is 0.5-1.5: 0.5-1.5: 0.5-1.5: 0.1-0.5: 0.1-0.5: 0.1 to 0.5.

8. The fermentation method of gibberellic acid GA3 ≧ 7.5g/L according to any one of claims 1-7, wherein the air flow rate is variable air flow rate of average air flow rate in the early stage of fermentation < average air flow rate in the middle stage of fermentation > average air flow rate in the late stage of fermentation.

9. The fermentation method of gibberellic acid GA3 ≧ 7.5g/L as claimed in claim 8, wherein the early stage of fermentation is a fermentation delay period and a logarithmic growth period, the middle stage of fermentation is a stationary period, and the late stage of fermentation is a decline period.

10. The fermentation method of gibberellic acid GA3 ≧ 7.5g/L according to claims 1-8, wherein the average air flow rate before fermentation is 3500-³Flow rate 3000-³H; average air flow rate of 4200-4400Nm in middle fermentation period³/h, air flow rate 3500-4600Nm³H; average air flow 3900-³/h。

Technical Field

The application relates to the field of gibberellic acid, in particular to a fermentation method of gibberellic acid GA3 ≧ 7.5 g/L.

Background

Gibberellic acid is a secondary metabolite obtained by fermentation culture and metabolism of a kind of gibberellin, 116 kinds of gibberellin are separated, identified and named, wherein GA3 is the most widely applied in agriculture, and plays a great role in agricultural production in China.

GA₃Has very high plant regulating activity and obvious regulating effect on the growth and development of various crops. In the north, GA₃The gibberellic acid solution with a certain concentration is sprayed in the flowering phase, so that the fruit setting rate of fruit trees can be effectively improved, the fruit growth and development are promoted, and the fruit yield is improved by about 20-30%. In the south, gibberellic acid is mainly applied to hybrid rice seed production, and male and female parents of rice are regulated to bloom at the same time, so that the pollination rate of the female parents is greatly improved, the yield of hybrid rice seeds is further improved, the production cost of the seeds is reduced, and the burden of farmers is reduced.

CN201810048112.2 discloses a fermentation method of gibberellic acid, wherein the fermentation method discloses a fermentation method, and a fermentation medium of the fermentation method consists of the following components: 10-40 g/L of corn protein powder; 1-10 g/L potassium dihydrogen phosphate; 5-30 g/L of a small-molecular organic carbon source; 0.5-5 g/L of vegetable oil; 0.5-2 g/L magnesium sulfate; 0.5-2 g/L of ammonium sulfate; 0.1-1 g/L of trace elements; the air flow rate of the fermentation is 2000-3000 Nm³The fermentation temperature is 28-30 ℃; the fermentation time is 8-10 days, and the highest GA of the fermentation method₃Up to 5372ppm, but still further GA improvement is required₃The content of (a).

Disclosure of Invention

In view of the above-mentioned drawbacks, in one aspect, the present application provides a fermentation method of gibberellic acid, in which GA is fermented₃The content of the gibberellic acid is not less than 7.5 g/L.

The technical scheme is as follows: a fermentation method of gibberellic acid GA3 ≧ 7.5g/L for high-yield gibberellic acid fermentation, transferring gibberellic acid seed liquid into a fermentation tank for fermentation, and is characterized in that: the air flow rate of the fermentation method is variable speed air flow rate, and the fermentation culture medium of the fermentation method comprises:

1-9 parts of corn protein powder;

1-10 parts by weight of monopotassium phosphate;

1-30 parts by weight of a small-molecular organic carbon source;

0.5-5 parts by weight of vegetable oil;

0.5-2 parts by weight of magnesium sulfate;

0.5-2 parts by weight of ammonium sulfate;

0.1-1 part by weight of trace elements;

1-10 parts by weight of soybean protein;

1-9 parts of wheat bran; and

0.001-0.5 weight portion of tween.

In one or more specific embodiments of the present application, the fermentation temperature in the fermentation is 28 ℃ to 30 ℃, the fermentation time is 8 to 10 days, and the fermentation pressure is 0.01 to 0.05 MPa.

In one or more specific embodiments of the present application, the small molecule organic carbon source is sucrose or glucose.

In one or more specific embodiments of the present application, the small molecule organic carbon source is sucrose and glucose.

In one or more specific embodiments of the present application, the ratio of sucrose: the weight ratio of glucose is 1-2: 2 to 1.

In one or more specific embodiments herein, the trace elements are ferrous sulfate, zinc sulfate, manganese sulfate, sodium molybdate, copper sulfate and cobalt chloride.

In one or more specific embodiments of the present application, the weight ratio of the ferrous sulfate, the zinc sulfate, the manganese sulfate, the sodium molybdate, the copper sulfate and the cobalt chloride is 0.5-1.5: 0.5-1.5: 0.5-1.5: 0.1-0.5: 0.1-0.5: 0.1 to 0.5.

In one or more specific embodiments of the present application, the air flow rate is a variable speed air flow rate of average air flow rate in a pre-fermentation stage < average air flow rate in a middle fermentation stage > average air flow rate in a post-fermentation stage.

In one or more specific embodiments of the present application, the early fermentation stage is a fermentation lag phase and a logarithmic growth phase, the middle fermentation stage is a stationary phase, and the late fermentation stage is a decline phase.

In one or more specific embodiments of the present application, the pre-fermentation average air flow rate is 3500-³Flow rate 3000-³H; average air flow rate of 4200-4400Nm in middle fermentation period³/h, air flow rate 3500-4600Nm³H; average air flow 3900-³/h。

The invention principle and the beneficial effects are as follows:

the present application surprisingly achieves GA by improving air flow and fermentation medium in fermentation, and combining the two₃The content of (b) is not less than 7.5 g/L.

Detailed Description

The present application will be further explained below.

Example 1 preparation of seed liquid before fermentation.

Preparing seed solution before fermentation according to CN201810048112.2 example 1.1-1.3, wherein GA₃The strains, the process and the culture medium are completely the same.

The test data of the prepared seed liquid are shown in the following table 1.

TABLE 1 Pre-fermentation seed liquid assay data

The seed solutions obtained in example 1 were subjected to the fermentation liquid preparation in the following examples 2 to 12, respectively.

In examples 2 to 12, the concentrations and the amounts of the seed solutions were equal.

Example 2

The seed liquid prepared in example 1 was fermented according to the fermentation method 1.4 in example 1 of CN201810048112.2 to obtain a fermentation liquid, and during the fermentation process, samples were taken respectively for detection, and the results are shown in table 2 below.

The fermentation process comprises the following steps:

fermentation volume: the volume is fixed to 82t, the volume after the extinction is 90t, and the volume after the rotation is 100 t.

Fermentation temperature: 29. + -. 0.2 ℃.

Operating the tank pressure: 0.035 MPa.

Air flow rate: 2200Nm³/h。

Controlling the pH: after the operation is started, the PH is gradually increased, the PH is increased by 0.1/half hour each time, and the PH is controlled to be 5.0-5.2 by adding ammonia water in a flowing mode.

And (3) supplementary material control: the dissolved oxygen rebounds by 20 percent, and the automatic feeding of glucose, salad oil and glucose (45 percent) solution is started: oil (3.2L:1L), dissolved oxygen control point 20% -30%.

And (3) fermentation period: 9 days (total fermentation time was 198 hours, since the time of transfer to the fermenter was 6 hours in the afternoon, day 1 in the fermenter).

TABLE 2 fermentation broth assay data

In this example, the fermentation medium is shown in Table 3 below.

TABLE 3 fermentation Medium composition content and sources

Example 3

This example was the same as example 2 except that the air flow rate was different.

Air flow of the present embodimentIn an amount of 4600Nm³/h。

In the fermentation process of this example, samples were taken for detection, and the results are shown in Table 4 below.

TABLE 4 fermentation broth assay data

Example 4

This example was the same as example 2 except that the air flow rate was different.

In the fermentation process of this example, samples were taken for detection, and the results are shown in Table 5 below.

TABLE 5 fermentation broth assay data

In this example, the air flow process parameters are shown in Table 6 below.

TABLE 6 air flow Process parameters

Example 5

This example was the same as example 2 except that the air flow rate was different.

In the fermentation process of this example, samples were taken for detection, and the results are shown in Table 7 below.

TABLE 7 fermentation broth assay data

In this example, the air flow process parameters are shown in Table 8 below.

TABLE 8 air flow Process parameters

Example 6

This example was the same as example 2 except that the air flow rate was different.

In the fermentation process of this example, samples were taken for detection, and the results are shown in Table 9 below.

TABLE 9 fermentation broth assay data

In this example, the air flow process parameters are shown in Table 10 below.

TABLE 10 air flow Process parameters

Example 7

This example is identical to example 2, except that the fermentation medium is different.

In the fermentation process of this example, samples were taken for detection, and the results are shown in Table 11 below.

TABLE 11 fermentation broth assay data

In this example, the fermentation media are shown in Table 12 below.

TABLE 12 fermentation Medium composition content and sources

Example 8

This example is identical to example 2, except that the fermentation medium is different.

In the fermentation process of this example, samples were taken for detection, and the results are shown in Table 13 below.

TABLE 13 fermentation broth assay data

In this example, the fermentation media are shown in Table 14 below.

TABLE 14 fermentation Medium composition content and sources

Example 9

This example is identical to example 2, except that the fermentation medium is different.

In the fermentation process of this example, samples were taken for detection, and the results are shown in Table 15 below.

TABLE 15 fermentation broth assay data

In this example, the fermentation media are shown in Table 16 below.

TABLE 16 fermentation Medium composition content and sources

Example 10

This example is identical to example 2, except that the air flow rate and the fermentation medium were different.

In this example, the fermentation medium was the fermentation medium of example 9.

The air flow rate is the air flow rate of example 4 (i.e., air flow process parameters of table 6).

In the fermentation process of this example, samples were taken for detection, and the results are shown in Table 17 below.

TABLE 17 fermentation broth assay data

Example 11

This example was identical to example 2 except that the air flow rate and the fermentation medium were different.

In this example, the fermentation medium was the fermentation medium of example 9.

The air flow rate is the air flow rate of example 5 (i.e., air flow process parameters of Table 8).

In the fermentation process of this example, samples were taken separately for detection, and the results are shown in Table 18 below.

TABLE 18 fermentation broth assay data

Example 12

This example was identical to example 2 except that the air flow rate and the fermentation medium were different.

In this example, the fermentation medium was the fermentation medium of example 9.

The air flow is the air flow of example 6 (i.e. table 10 air flow process parameters).

In the fermentation process of this example, samples were taken for detection, and the results are shown in Table 19 below.

TABLE 19 fermentation broth assay data

In this application, unless otherwise specified, all are prior art.

The above description is only a preferred embodiment of the present application and is not intended to limit the present application, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.