阅读说明：本技术 硫酸新霉素发酵清洁生产方法 (Neomycin sulfate fermentation clean production method ) 是由 谢飞 胡珂 崔国英 谭兴龙 王强 江兵 于 2019-11-26 设计创作，主要内容包括：一种硫酸新霉素发酵清洁生产方法,该方法包括以下步骤：步骤1：制得斜面孢子悬浮液；步骤2：制得种子液；步骤3：制得发酵液,发酵过程中通过补加补料培养基及硫酸铵,使新霉素的积累量达到发酵终点要求后终止发酵,制得发酵液,其中,发酵基础培养基包括2-10% (W/V)的麦芽糊精,发酵基础培养基中不含有米粉、淀粉及α-淀粉酶；补料培养基包括20-45% (W/V)的麦芽糊精,补料培养基中不含有米粉、淀粉及及α-淀粉酶；步骤4：将发酵液放入吸附罐,然后投入树脂对新霉素进行吸咐,即制得硫酸新霉素成品。本发明提供的硫酸新霉素发酵清洁生产方法,可以解决污染大、能耗高的问题,提高碳源类物质的利用率,实现清洁生产。(A neomycin sulfate fermentation clean production method comprises the following steps of 1, preparing a slope spore suspension, 2, preparing a seed liquid, 3, preparing a fermentation liquid, wherein in the fermentation process, a supplemented culture medium and ammonium sulfate are supplemented to stop fermentation after the accumulation of neomycin reaches a fermentation end point requirement, so that the fermentation liquid is prepared, wherein a fermentation basic culture medium comprises 2-10% (W/V) of maltodextrin, rice flour, starch and α -amylase are not contained in the fermentation basic culture medium, the supplemented culture medium comprises 20-45% (W/V) of maltodextrin, rice flour, starch and α -amylase are not contained in the supplemented culture medium, and 4, the fermentation liquid is placed in an adsorption tank, and resin is added to adsorb neomycin, so that a neomycin sulfate finished product is prepared.)

1. A neomycin sulfate fermentation clean production method is characterized by comprising the following steps:

step 1: inoculating sand soil spores to a slant, and culturing at 28 +/-1 ℃ for 4-8 days to obtain a slant spore suspension;

step 2: inoculating the slant spore suspension into a seeding tank filled with a seed culture medium, and culturing for 35-40 h to obtain a seed solution;

and step 3: inoculating the seed solution into a fermentation tank filled with a fermentation basal culture medium, fermenting for 150-180 h to obtain a fermentation broth, supplementing a supplementary culture medium and ammonium sulfate in the fermentation process to ensure that the fermentation is stopped after the accumulation of the neomycin meets the fermentation end point requirement to obtain the fermentation broth,

wherein the fermentation basal culture medium comprises 2-10% W/V maltodextrin, and the fermentation basal culture medium does not contain rice flour and α -amylase;

the feed medium comprises 20-45% W/V maltodextrin, and the feed medium does not contain rice flour, starch and α -amylase;

and 4, step 4: and (3) putting the fermentation liquor into an adsorption tank, and then adding resin to adsorb neomycin to obtain a neomycin sulfate finished product.

2. The neomycin sulfate fermentation clean production method of claim 1, characterized in that: in step 3, the fermentation basal medium comprises 5% W/V maltodextrin.

3. The neomycin sulfate fermentation clean production method of claim 1, characterized in that: in step 3, the feed medium included 35% W/V maltodextrin.

Technical Field

The invention relates to the technical field of biological fermentation, in particular to a neomycin sulfate fermentation clean production method.

Background

Rice flour and starch are used as fermentation carbon sources in a large amount in neomycin sulfate fermentation production, particularly grain raw material rice flour, although the nutrient components are rich, the carbohydrate accounts for 70-80%, and the protein and amino acid account for about 10%, the grain size is large, the rice flour is insoluble in water, and the utilization rate of microbial strains is low; meanwhile, neomycin sulfate production mainly pursues improvement of fermentation level, a large amount of rice flour and starch hydrolysis sugar liquid are supplemented, unit consumption and conversion rate of saccharides are rarely considered, and more residues are left after fermentation is finished, so that main pollutant indexes are high, and sewage treatment pressure is high and treatment cost is high.

After the fermentation basal culture medium and the feeding culture medium use rice flour and starch, only a sterilization mode of actual sterilization can be adopted to ensure the sterility of the culture medium due to the high solid content, and the steam consumption is larger than that of a continuous sterilization mode, so that the energy consumption is high.

Disclosure of Invention

The technical problem to be solved by the invention is to provide a neomycin sulfate fermentation clean production method, which can solve the problems of high pollution and high energy consumption, improve the utilization rate of carbon source substances, reduce the material unit consumption, further improve the fermentation unit level, simultaneously reduce the energy consumption in the production process, reduce the ammonia nitrogen emission concentration by about 25 percent when the fermentation is finished, and realize clean production.

In order to solve the technical problems, the technical scheme adopted by the invention is as follows: a neomycin sulfate fermentation clean production method comprises the following steps:

step 1: inoculating sand soil spores to a slant, and culturing at 28 +/-1 ℃ for 4-8 days to obtain a slant spore suspension;

step 2: inoculating the slant spore suspension into a seeding tank filled with a seed culture medium, and culturing for 35-40 h to obtain a seed solution;

and step 3: inoculating the seed solution into a fermentation tank filled with a fermentation basal culture medium, fermenting for 150-180 h to obtain a fermentation broth, supplementing a supplementary culture medium and ammonium sulfate in the fermentation process to ensure that the fermentation is stopped after the accumulation of the neomycin meets the fermentation end point requirement to obtain the fermentation broth,

wherein the fermentation basal culture medium comprises 2-10% (W/V) of maltodextrin, and does not contain rice flour and α -amylase;

the formula ratio of the fermentation basal medium after replacing rice flour and α -amylase by maltodextrin is shown in table 1:

TABLE 1

The feed medium comprises 20-45% W/V maltodextrin, and the feed medium does not contain rice flour, starch and α -amylase;

the formula ratio of the feed medium after replacing rice flour, starch and α -amylase with maltodextrin was changed as shown in Table 1:

TABLE 2

And 4, step 4: and (3) putting the fermentation liquor into an adsorption tank, and then adding resin to adsorb neomycin to obtain a neomycin sulfate finished product.

Preferably, in step 3, the fermentation basal medium comprises 5% (W/V) maltodextrin.

Preferably, in step 3, the feed medium comprises 35% (W/V) maltodextrin.

The neomycin sulfate fermentation clean production method provided by the invention has the following beneficial effects:

1. the culture medium uses maltodextrin to replace rice flour and starch, and the culture medium is easy to be utilized by microorganisms because the culture medium contains more monosaccharide and disaccharide; especially in the middle and later stages of fermentation, when the amylase secreted by the thalli per se is reduced, the anabolism requirement of neomycin can still be ensured, so that the unit consumption of main carbon source substances is reduced by 10.77 percent, and the purpose of 'making the best of things' is achieved, as shown in Table 3

The comparison of the clean production process conditions of the 50T fermenter with the existing process conditions is shown in Table 3:

TABLE 3

2. The average fermentation level is improved by 4.56 percent, the average yield of single-tank fermentation liquor is improved by 10.86 percent, and the labor production efficiency is improved; under the condition of unchanged production period, the specific energy consumption level is reduced.

3. When the fermentation is finished, the ammonia nitrogen index in the fermentation liquor is relatively stable, the average level of the ammonia nitrogen index is greatly reduced by 25.03 percent compared with the average level of the ammonia nitrogen index in the prior art, the environmental protection pressure of enterprises is relieved, and the pollutant treatment cost is reduced.

4. The product has stable quality, and all indexes of the product reach or exceed the standards of neomycin EP9.0 pharmacopeia (Table 4)

TABLE 4

5. The energy consumption cost is obviously reduced: although rice flour is crushed and sieved in a traditional production process of neomycin sulfate, the rice flour is still large in granularity and insoluble in water, and in order to ensure the sterility of a culture medium, only a sterilization mode of actual sterilization can be adopted for sterilization, so that the using amount of steam is large and the cost is high. After the clean production process uses the maltodextrin to replace rice flour, because the maltodextrin is easy to be watered and the solid content in the culture medium is obviously reduced, a continuous sterilization mode is adopted, and the steam consumption is saved by about 30 percent.

6. The quantity of the culture medium materials is reduced, and the risk of errors of ingredients is reduced; the preparation process does not need heat preservation liquefaction, simplifies the batching process, greatly reduces dust pollution and improves the batching efficiency.

7. The maltodextrin is used as an industrial product, has definite national standard for quality, is easier to control than agricultural and sideline product rice flour, and is beneficial to stable production.

Detailed Description