阅读说明：本技术 一种高诱食性和适口性的有机微量元素及其制备方法 (Organic trace element with high food calling property and palatability and preparation method thereof ) 是由 郑理想 高小创 孙中超 杨旭 夏志生 于 2021-08-11 设计创作，主要内容包括：本发明一种高诱食性和适口性的有机微量元素及其制备方法,该制备方法包括如下步骤：将生香酵母菌株经过一级种子培养、二级种子扩培和耗氧扩培后,在厌氧环境下培养3-4小时；将配制好的高浓度无机盐金属离子溶液不断地滴注在厌氧培养后的发酵液中,然后在厌氧条件下进行螯合2-3小时；然后升温,在一定条件下生香酵母菌体充分自溶；采用薄膜干燥法对自溶后的生香酵母菌液进行干燥,然后加工制备成有机微量元素产品。本发明提供的方法生产过程中无需离心、浓缩、抽提洗涤等过程,无有机溶剂排放产生,工艺相对简单,过程成本低,适合大规模工业化生产。此外,制备的产品解决了当前有机微量元素适口性低的问题,具有较大的应用前景与市场空间。(The invention relates to an organic microelement with high food calling and palatability and a preparation method thereof, wherein the preparation method comprises the following steps: after primary seed culture, secondary seed expansion culture and oxygen consumption expansion culture, the aroma-producing yeast strain is cultured for 3 to 4 hours in an anaerobic environment; continuously dripping the prepared high-concentration inorganic salt metal ion solution into fermentation liquor after anaerobic culture, and then chelating for 2-3 hours under anaerobic conditions; then heating, and fully autolyzing aroma-producing yeast thalli under certain conditions; and drying the autolyzed aroma-producing yeast liquid by adopting a film drying method, and then processing to prepare the organic trace element product. The method provided by the invention does not need processes such as centrifugation, concentration, extraction and washing and the like in the production process, does not discharge organic solvent, has relatively simple process and low process cost, and is suitable for large-scale industrial production. In addition, the prepared product solves the problem of low palatability of the current organic trace elements, and has a wide application prospect and a wide market space.)

1. A preparation method of organic trace elements with high food calling property and palatability is characterized by comprising the following steps:

step one, after primary seed culture, secondary seed expansion culture and oxygen consumption expansion culture, culturing the saccharomyces cerevisiae strain for 3-4 hours in an anaerobic environment;

continuously dripping the prepared high-concentration inorganic salt metal ion solution into the fermentation liquor after anaerobic culture in the step one, and then chelating for 2-3 hours under an anaerobic condition;

step three, after chelation, heating, and fully autolyzing the aroma-producing yeast cells under the conditions of pH6.5-7.5 and the rotation speed of 100-150 rpm;

and step four, drying the autolyzed aroma-producing yeast liquid by adopting a film drying method, and then crushing, subpackaging and packaging to prepare the organic trace element product.

2. The method according to claim 1, wherein the culture medium used for the primary seed culture and the secondary seed expansion culture comprises the following components: 10-15g/L of glucose, 8-10g/L of beef extract powder and 8-10g/L of yeast extract.

3. The method according to claim 1, wherein the amount of inoculated strain of Saccharomyces cerevisiae in the primary seed culture is 1-2%, the culture temperature is 37-40 ℃, the rotation speed is 50-100rpm, and the culture time is 16-20 hours.

4. The method according to claim 1, wherein the amount of inoculated strain of Saccharomyces cerevisiae in the secondary seed culture is 5-10%, the culture temperature is 37-40 ℃, the rotation speed is 100-150rpm, and the culture time is 10-12 hours.

5. The method according to claim 1, wherein the amount of inoculated strain of Saccharomyces cerevisiae is 5-20%, the culture temperature is 37-40 ℃, the rotation speed is 100-150rpm, the aeration rate is 0.2-0.4vvm/h, and the culture time is 6-8 hours.

6. The method according to claim 1, wherein the inorganic salt metal ion solution in the second step is prepared by the following method: preparing a 2% sodium carboxymethylcellulose solution, and dissolving metal inorganic salt in the 2% sodium carboxymethylcellulose solution to ensure that the concentration of metal ions reaches 20% -30%.

7. The method according to claim 1, wherein in the second step, the inorganic salt metal ion solution is dripped in an amount of 10 to 20% by volume of the fermentation broth at a dripping speed of 10 to 15L/hr.

8. The process according to claim 1, wherein the temperature at the time of chelation in the second step is 50 to 55 ℃, the rotation speed is 0 to 50rpm, the pH is 5.5 to 6.5, and the chelation time is 2 to 3 hours.

9. The process according to claim 1, wherein the temperature is raised to 60 to 65 ℃ in the third step, and the autolysis time of the aroma-producing yeast is 1 to 2 hours.

10. An organic trace element produced by the production method according to any one of claims 1 to 9.

Technical Field

The invention relates to the field of organic trace elements, in particular to an organic trace element with high food calling property and palatability and a preparation method thereof.

Background

The trace elements are indispensable nutrient elements in the normal growth and metabolism process of livestock and poultry bodies, and the trace elements which are circulated in the market at present mainly comprise inorganic salt trace elements, organic acid complex trace elements, polysaccharide complex trace elements, amino acid chelate trace elements and protein chelate trace elements. Wherein the inorganic salt trace elements and the organic acid trace elements are gradually replaced by the organic trace elements due to various defects of uneven mixing, poor stability, easy deliquescence, easy oxidation, low absorption and utilization rate and the like. The organic trace elements are formed by complexing or chelating polysaccharide, amino acid, protein and metal trace elements, have high stability and digestibility, and become the mainstream direction of current research, development, production and application. The preparation method comprises the steps of preparing chelating substrate solution such as polysaccharide, amino acid and the like by dissolving or enzymolysis, adding inorganic trace elements, reacting for several hours at high temperature to obtain complex solution, centrifuging, concentrating, extracting and washing, mixing protective agents, drying and crushing. The prior patent technologies of the invention comprise 'an enzymolysis composition, a method for enzymolysis of soybean meal and a preparation method of an organic trace element feed additive-CN 111528342A', 'a biological organic mineral compound additive for reducing cracked eggs of eggs and a use method-CN 111296640A', 'cassava residue organic trace element, a preparation method and an animal feed-CN 111280306A', 'a biological organic mineral compound additive for reducing dark spots of eggs and a use method-CN 111296641A', 'a small peptide production process rich in one or more trace elements-CN 111154825A' and the like.

Compared with the traditional product, the product invented by the prior patent technology is improved to a certain extent, but still has the defects of complex production process, generation of organic solvent waste liquid and the like. In addition, the development of the organic trace element products mainly focuses on improving the stability and the digestibility, the improvement of the palatability is concerned a little, and the organic trace element products and the patent technology inventions focusing on the palatability and the palatability are not seen so far. For the livestock and poultry breeding industry, poor palatability is a key factor influencing animal feeding and is an important obstacle for market promotion of organic trace elements. The development of organic trace element products with high food calling property and palatability is an urgent need of the current livestock and poultry breeding and feed industries, and has important commercial value and wide application prospect.

Disclosure of Invention

Aiming at the defects of the prior art and filling the blank of products in the market, the invention provides an organic trace element with high food calling property and palatability and a preparation method thereof.

In order to achieve the purpose, the invention adopts the following technical scheme:

the first aspect of the invention provides a preparation method of organic trace elements with high food calling property and palatability, which comprises the following steps:

step one, after primary seed culture, secondary seed expansion culture and oxygen consumption expansion culture, culturing the saccharomyces cerevisiae strain for 3-4 hours in an anaerobic environment;

continuously dripping the prepared high-concentration inorganic salt metal ion solution into the fermentation liquor after anaerobic culture in the step one, and then chelating for 2-3 hours under an anaerobic condition;

step three, after chelation, heating, and fully autolyzing the aroma-producing yeast cells under the conditions of pH6.5-7.5 and the rotating speed of 100-150 rpm;

and step four, drying the autolyzed aroma-producing yeast liquid by adopting a film drying method, and then crushing, subpackaging and packaging to prepare the organic trace element product.

Further, the culture medium adopted by the primary seed culture and the secondary seed expansion culture comprises the following components: 10-15g/L of glucose, 8-10g/L of beef extract powder and 8-10g/L of yeast extract.

Further, in the primary seed culture, the inoculation amount of the aroma-producing yeast strain is 1-2%, the culture temperature is 37-40 ℃, the rotation speed is 50-100rpm, and the culture time is 16-20 hours.

Further, in the secondary seed culture, the inoculation amount of the aroma-producing yeast strain is 5-10%, the culture temperature is 37-40 ℃, the rotation speed is 100-150rpm, and the culture time is 10-12 hours.

Furthermore, in the oxygen-consuming expanding culture, the inoculation amount of the aroma-producing yeast strain is 5-20%, the culture temperature is 37-40 ℃, the rotation speed is 100-150rpm, the ventilation amount is 0.2-0.4vvm/h, and the culture time is 6-8 hours.

Further, the culture medium adopted by the oxygen-consuming culture extension comprises the following components: 2-5g/L of glucose, 7-12g/L of corn flour, 100 g/L of soybean meal and 150g/L of beef extract powder, and 3-8g/L of beef extract powder.

Further, the preparation method of the inorganic salt metal ion solution in the second step is as follows: preparing a 2% sodium carboxymethylcellulose solution, and dissolving metal inorganic salt in the 2% sodium carboxymethylcellulose solution to ensure that the concentration of metal ions reaches 20% -30%.

Further, in the second step, the dripping amount of the inorganic salt metal ion solution is 10-20% of the volume of the fermentation liquid, and the dripping speed is 10-15L/h.

Further, the temperature of chelation in the second step is 50-55 ℃, the rotating speed is 0-50rpm, the pH is 5.5-6.5, and the chelation time is 2-3 hours.

Further, in the third step, the temperature is raised to 60-65 ℃, and the autolysis time of the aroma-producing yeast is 1-2 hours.

Furthermore, the temperature adopted in the drying in the fourth step is 90-95 ℃, and the drying time is 3-5 minutes.

The second aspect of the invention is to provide the organic trace element prepared by the preparation method.

By adopting the technical scheme, compared with the prior art, the invention has the following technical effects:

the method provided by the invention can produce organic trace element products with high palatability and palatability, which have the advantages of hundreds of aromatic substances, chelating rate of 99%, crude protein content of 32% and small peptide content of 15%, and the production process does not need the processes of centrifugation, concentration, extraction, washing and the like, and does not discharge organic solvents. In addition, the prepared organic trace elements solve the problem of low palatability of the existing organic trace elements, and have a wide application prospect and a wide market space.

Drawings

FIG. 1 is a process flow diagram of a method for preparing organic trace elements with high palatability and palatability according to an embodiment of the invention;

FIG. 2 is a GC-MS analysis spectrum of an organic trace element according to an embodiment of the present invention;

FIG. 3 is an HPLC analysis chart of organic trace elements according to an embodiment of the present invention;

FIG. 4 shows the results of experimental verification of animal husbandry in accordance with an embodiment of the present invention; wherein, the graphs A to C respectively show the measurement results of the daily average feed intake, the daily average weight gain and the feed-meat ratio;

fig. 5 is a graph showing the measurement result of the ion content in the serum of the growing piglets after being fed according to the embodiment of the invention.

Detailed Description

The invention provides an organic trace element with high food calling property and palatability and a preparation method thereof, comprising a high-activity fermentation strain, a strain culture process method and a trace element chelation production process method. The production fermentation strain selects a high-performance aroma-enhancing yeast strain, and fermentation strains with high adsorbability, high chelation and high palatability, optimal fermentation medium components, optimal feeding fermentation process, optimal fermentation culture conditions, optimal adsorption and chelation process, optimal product secretion process and large-scale amplification fermentation process parameters are obtained through single factor test, Plackett-Burman test, steepest climbing test and response surface optimization.

In a preferred embodiment of the present invention, the method for preparing organic trace elements with high palatability and palatability comprises the following steps:

step one, after primary seed culture, secondary seed expansion culture and oxygen consumption expansion culture, culturing the saccharomyces cerevisiae strain for 3-4 hours in an anaerobic environment;

continuously dripping the prepared high-concentration inorganic salt metal ion solution into the fermentation liquor after anaerobic culture in the step one, and then chelating for 2-3 hours under an anaerobic condition;

step three, after chelation, heating, and fully autolyzing the aroma-producing yeast cells under the conditions of pH6.5-7.5 and the rotating speed of 100-150 rpm;

and step four, drying the autolyzed aroma-producing yeast liquid by adopting a film drying method, and then crushing, subpackaging and packaging to prepare the organic trace element product.

In a preferred embodiment of the present invention, the culture medium used for the primary seed culture and the secondary seed expansion culture comprises the following components: 10-15g/L of glucose, 8-10g/L of beef extract powder and 8-10g/L of yeast extract.

In a preferred embodiment of the present invention, in the primary seed culture, the inoculation amount of the strain of the aroma-producing yeast is 1-2%, the culture temperature is 37-40 ℃, the rotation speed is 50-100rpm, and the culture time is 16-20 hours.

In a preferred embodiment of the present invention, in the secondary seed culture, the inoculation amount of the strain of the Saccharomyces cerevisiae is 5-10%, the culture temperature is 37-40 ℃, the rotation speed is 100-150rpm, and the culture time is 10-12 hours.

In a preferred embodiment of the present invention, in the oxygen-consuming expanding culture, the inoculation amount of the strain of the Saccharomyces cerevisiae is 5-20%, the culture temperature is 37-40 ℃, the rotation speed is 100-150rpm, the ventilation amount is 0.2-0.4vvm/h, and the culture time is 6-8 hours.

In a preferred embodiment of the present invention, the culture medium used in the oxygen-consuming culture includes the following components: 2-5g/L of glucose, 7-12g/L of corn flour, 100 g/L of soybean meal and 150g/L of beef extract powder, and 3-8g/L of beef extract powder.

In a preferred embodiment of the present invention, the preparation method of the inorganic salt metal ion solution in the second step is as follows: preparing a 2% sodium carboxymethylcellulose solution, and dissolving metal inorganic salt in the 2% sodium carboxymethylcellulose solution to ensure that the concentration of metal ions reaches 20% -30%.

In a preferred embodiment of the present invention, in the second step, the dropping amount of the inorganic salt metal ion solution is 10-20% of the volume of the fermentation liquid, and the dropping speed is 10-15L/h.

In a preferred embodiment of the present invention, the temperature at the time of chelation in the second step is 50 to 55 ℃, the rotation speed is 0 to 50rpm, the pH is 5.5 to 6.5, and the chelation time is 2 to 3 hours.

In a preferred embodiment of the present invention, in step three, the temperature is raised to 60-65 ℃ and the autolysis time of the aroma-producing yeast cells is 1-2 hours.

In a preferred embodiment of the present invention, the drying in the fourth step is carried out at a temperature of 90 to 95 ℃ for a drying time of 3 to 5 minutes.

The organic trace element product with good palatability and high chelating rate is obtained by seed culture, fermentation culture, feeding culture, trace element adsorption and chelation, product secretion and release, drying and crushing. The trace element product has high chelating rate up to 99% through the adsorption and chelating of high performance aroma-enhancing yeast to trace elements and through unique secretion and release process, and contains a great amount of yeast nucleotide, phenol, aldehyde, ketone and other aroma-producing substances in hundreds, so that the trace element product has good palatability and palatability.

The organic trace element product of the invention uses high-performance aroma-enhancing yeast fermentation thalli to adsorb and enrich chelated trace elements, and is rich in various feeding-inducing metabolites. The palatability of the product is greatly improved through screening of fermentation strains and a fermentation medium, optimization of fermentation conditions, optimization of a flavor-enhancing and aroma-increasing process, optimization of an element adsorption and chelation process, a product secretion and release process and optimization of large-scale fermentation process parameters. The aroma-increasing yeast strains screened by the product can produce various aromatic metabolites, have extremely strong metal adsorption and enrichment capacity, and have the advantages that after secondary seed amplification culture, the thalli are greatly increased in an oxygen-consuming fermentation stage, and after the secondary seed amplification culture, the yeast metabolism is performed to produce nearly hundreds of aroma substances such as alcohols, aldehydes, phenols, ketones and the like, and the palatability of the aroma-increasing yeast strains is obviously higher than that of the existing organic trace products. The invention continuously instills metal ion solution at a certain speed, makes yeast thalli adsorb metal ions under the anaerobic condition, and the metal ions and intracellular small peptide form chelate and are stored in cells, the stability of the chelate product is high, and the chelate rate reaches 99%. The yeast nucleotide has good delicate flavor effect and can obviously attract food. In the product secretion and release stage, the temperature is raised to 65 ℃ for full autolysis of yeast thalli, so that intracellular yeast nucleic acid is also fully released while releasing chelate compounds such as yeast metalloprotein, yeast metallopolysaccharide and the like, and the food calling property and the palatability of the product are further improved.

The present invention will be described in detail and specifically with reference to the following examples and drawings so as to provide a better understanding of the invention, but the following examples do not limit the scope of the invention.

In the examples, the conventional methods were used unless otherwise specified, and reagents used were those conventionally commercially available or formulated according to the conventional methods without specifically specified.

Example 1

Referring to fig. 1, this example provides a method for preparing organic trace elements with high palatability and palatability, using aroma-producing yeast, comprising the following steps:

step one, primary seed culture:

primary seed culture medium: 600g of glucose, 480g of beef extract powder, 480g of yeast extract and 60L of deionized water;

the culture conditions are as follows: the inoculation amount is 1-2%, the temperature is 37-40 ℃, the rotating speed is 50-100rpm, and the culture time is 16-20 h.

Step two, expanding and culturing secondary seeds:

secondary seed culture medium: 6000g of glucose, 4800g of beef extract powder, 4800g of yeast extract and 600L of deionized water;

the culture conditions are as follows: the inoculation amount is 5-10%, the temperature is 37-40 ℃, the rotation speed is 100 and 150rpm, and the culture time is 10-12 h.

Step three, oxygen-consuming and expanding culture of thalli:

aerobic expanding culture medium: 12.6kg of glucose, 42kg of corn flour, 504kg of soybean meal, 21kg of beef extract powder and 4200L of deionized water;

the culture conditions are as follows: the inoculum size is 15-20%, the temperature is 37-40 ℃, the rotation speed is 100-.

Step four, flavor improvement and aroma enhancement: after the thalli are subjected to aerobic expanding culture for 6-8h, the ventilation is closed, and the thalli are cultured in an anaerobic environment for 3-4 h.

Step five, dripping, absorbing and enriching elements: preparing 20-30% of high-concentration inorganic salt metal ion solution, and then continuously dripping the metal ion solution at a certain speed, wherein the dripping amount is 10-20% of the volume of the fermentation liquid, and the dripping speed is 10-15L/h;

element absorption and enrichment chelation: anaerobic reaction at 50-55 deg.c, rotation speed of 0-50rpm, pH5.5-6.5 and chelating time of 2-3 hr.

Step six, autolysis: the temperature is 60-65 ℃, the pH6.5-7.5, the rotating speed is 100-.

Step seven, drying the film: the drying temperature is 90-95 deg.C, and the drying time is 3-5 min.

Step eight, crushing, subpackaging and packaging: and crushing the dried product, subpackaging and packaging to prepare the organic trace element product.

Verification examples

1. Feeding habit analysis

The prepared organic trace element product was analyzed by GC-MS, and as shown in fig. 2 and table 1 below, the product contained 28 substances that were able to make animals enjoy and pleasure in eating.

Table 1 substances contained in the organic trace element products that make animals enjoy and pleasure eating them

The prepared organic trace element product is analyzed by HPLC, as shown in figure 3, the product contains a large amount of umami nucleotide, the contents of CMP, UMP, GMP, AMP and TMP are more than 600ppm, and the total yeast nucleotide is more than 3000 ppm.

2. Animal breeding experiment verification

The experimental scheme is as follows: 120 piglets of 110 days old weighed 50.0 + -0.5 kg initially, and were divided into a test group and a control group by a completely random block, each group had 3 replicates, and each replicate 20 piglets.

Control group: the product of the invention is not added; test groups: 0.1% of the product of the invention was added, with a test period of 30 days.

As can be seen from FIG. 4, compared with the control group, the feed intake of the patented product added with 0.1% of the feed additive is improved by 7.8%, the daily gain is increased by 16%, and the feed conversion ratio is reduced by 6.9%.

3. Absorbance analysis

The experimental scheme is as follows: 360 piglets growing up and growing up with 62 plus or minus 2kg of initial weight are selected and divided into 6 groups according to a complete random block, namely a group A (30ppm of inorganic copper), a group B (100ppm of inorganic copper), a group C (150ppm of inorganic copper), a group D (10ppm of the product of the invention), a group E (30ppm of the product of the invention) and a group F (60ppm of the product of the invention).

Each group had 3 replicates, each replicate 20 pigs.

The culture period is as follows: for 40 days.

Adopting an anterior vena cava to take blood, and using a disposable syringe to take 3mL blood sample of each test pig; centrifuging the blood sample, separating to obtain serum, and storing in a 1.5mL centrifuge tube; measuring the ion content in the serum by using an atomic absorption method; serum immunoglobulin content was determined using immunotransmission turbidimetry. The results are shown in FIG. 5 and Table 2 below.

TABLE 2 serum immunological index measurement results of raised pigs

As can be seen in FIG. 5, the higher level of ions in serum using the low dose of the product of the invention indicates a better absorption of the product of the invention.

As can be seen from Table 2, the serum immune indexes GLB (serum globulin), IgA (serum immunoglobulin A) and IgG (serum immunoglobulin G) of the experimental pig are higher by using the low-dose product of the invention, and the immune function of the body is reflected to be better, so that the product of the invention has high absorption rate in the livestock and poultry body.

The embodiments of the present invention have been described in detail, but the embodiments are merely examples, and the present invention is not limited to the embodiments described above. Any equivalent modifications and substitutions to those skilled in the art are also within the scope of the present invention. Accordingly, equivalent changes and modifications made without departing from the spirit and scope of the present invention should be covered by the present invention.