CN112094747B - Preparation method of kluyveromyces marxianus active dry yeast - Google Patents
Preparation method of kluyveromyces marxianus active dry yeast Download PDFInfo
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Abstract
The invention belongs to the technical field of microorganism live bacteria preparations, and particularly relates to a preparation method of kluyveromyces marxianus active dry yeast, which comprises the following steps: firstly, filtering Kluyveromyces marxianus fermentation liquor to obtain fresh yeast paste; re-dissolving the fresh yeast paste in a pretreatment agent solution to obtain a re-dissolved yeast suspension, and filtering to obtain pretreated yeast paste I; then, breaking up the pretreated yeast paste I, adding a protective agent, and continuously stirring for 20 to 30min to obtain pretreated yeast paste II; and finally extruding the pretreated yeast paste II into yeast wet particles, putting the yeast wet particles into a fluidized bed dryer for drying in two sections, drying the yeast wet particles for 5 to 15min at 50 to 65 ℃ in the first section, controlling the water content of the material to be 25 to 35 percent when the drying is finished, reducing the drying temperature to 35 to 42 ℃ in the second section until the water content of the material is 4 to 6 percent, and finishing the drying to obtain the kluyveromyces marxianus active dry yeast product. The invention not only can improve the viable count and the storage stability of the Kluyveromyces kluyveromyces lactis active dry yeast product, but also can reduce the production cost.
Description
Technical Field
The invention belongs to the technical field of microorganism live bacteria preparations, and particularly relates to a preparation method of kluyveromyces marxianus active dry yeast.
Background
With the rapid development of livestock breeding industry in China, microbial feed additives are paid much attention, and saccharomycetes is one of the common saccharomycetes. The microzyme belongs to facultative anaerobe, can survive and reproduce in aerobic and anaerobic states, has more nutrient substances, and is safe, rich and balanced in nutrition. Since the 40 s of the 20 th century, yeast was used in feed additives.
Kluyveromyces marxianus is food safety grade yeast, has been approved by the food safety certification of European Union and the FDA in the United states, and is also approved as a new food raw material in China. The cell wall components of mannan and glucomannan of Kluyveromyces marxianus, and enzyme system, protein, amino acid, nucleic acid, vitamins, trace elements, etc. produced by synthesis and metabolism have high bioactivity and nutrition. In addition, the active kluyveromyces marxianus can inhibit harmful bacteria in animal intestinal tracts, enhance the overall immunity of animals, promote the digestion and absorption of the animals on plants and lipid, and improve the utilization rate of feed. Therefore, kluyveromyces marxianus can be used as a novel feed additive microorganism for improving animal immunity and feed utilization rate.
Although kluyveromyces marxianus has a good application prospect as a beneficial feed additive, the kluyveromyces marxianus is not common in practical application, and the reason is that the kluyveromyces marxianus active dry yeast is difficult to industrially produce, particularly, on one hand, the production process cost is high, so that a feed factory and a culture terminal cannot accept the kluyveromyces marxianus freeze-dried powder, such as a preparation method of kluyveromyces marxianus freeze-dried powder for reducing serum cholesterol (CN 102524794A) applied by Liuhui et al of Beijing academy of agriculture and the preparation method of kluyveromyces marxianus freeze-dried powder (CN 1106159A) applied by Wang et al of Jiangsu university, the freeze-drying process is adopted to prepare live bacteria preparations, and the cost of the process cannot be accepted by the feed industry; another aspect is the prepared active dry fermentationThe viable count of the mother bacteria is low, and the viable count of the bacteria powder prepared in CN 102524794A is 2 × 10 9 CFU/g is about, and the viable count in the bacterial powder prepared in CN 11061616159A is 5.81 multiplied by 10 8 CFU/g, the number of live bacteria of common feeding yeast products on the market is generally 1 to 2 multiplied by 10 10 CFU/g, and the condition of synthesizing the production cost and the viable count of the product, the kluyveromyces marxianus freeze-dried powder prepared by the existing method has no feeding application value. Therefore, there is a need to design a new method for preparing kluyveromyces marxianus active dry yeast to overcome the above problems.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention aims to provide the preparation method of the kluyveromyces marxianus active dry yeast capable of realizing industrial production, which can prepare the kluyveromyces marxianus active dry yeast product with high viable count, has greatly reduced production cost compared with a freeze drying process, and is suitable for the additive of animal feed.
In order to achieve the aim, the technical scheme of the invention is a preparation method of kluyveromyces marxianus active dry yeast, which comprises the following steps:
1) Filtering kluyveromyces marxianus fermentation liquor obtained by high-density fermentation to obtain fresh yeast paste;
2) Preparing a pretreating agent into a pretreating agent solution with the concentration of 5-20% by using water; the main functions of the pretreatment agent are mainly two aspects, on one hand, yeast is protected from inactivation in the drying process, drying loss is reduced, on the other hand, the storage stability of dry yeast is improved, and the shelf life of products is prolonged;
3) Redissolving the fresh yeast paste in the step 1) in the pretreating agent solution in the step 2), and stirring for 0.5 to 2h at the temperature of 20 to 25 ℃ to obtain a redissolved yeast suspension;
4) Filtering the re-dissolved yeast suspension in the step 3) to obtain pretreated yeast paste I;
5) Scattering the pretreated yeast paste I by using a stirrer, adding a protective agent into the pretreated yeast paste I, and then continuously stirring for 20-30min to obtain pretreated yeast paste II; the protective agent has the main functions of reducing the death of yeast in the drying process, improving the viable bacteria yield of the product and simultaneously improving the storage stability of the product;
6) Extruding the pretreated yeast paste II into noodle-shaped yeast wet particles by using an extruder, then putting the yeast wet particles into a fluidized bed dryer for drying in two sections, drying at 50-65 ℃ for 5-15min in the first stage, controlling the water content of the material to be 25-35% after the drying is finished, reducing the drying temperature to 35-42 ℃ in the second stage until the water content of the material is 4-6%, and finishing the drying to obtain the Mackeuyveromyces marxianus active dry yeast product. The rapid drying of the moisture can be realized by adopting higher drying temperature in the first-stage drying process, the drying time is shortened, and meanwhile, the death of viable bacteria is less ensured by controlling the moisture at the end of drying to be not less than 25%; and in the second stage, the lower drying temperature is adopted, so that the moisture of the product is further reduced to a condition suitable for storage, the product is ensured to be suitable for storage, and the death of thalli in the drying process can be reduced by the lower drying temperature.
Further, in the step 5), the protective agent comprises an emulsifier and a colloidal substance which are mixed according to the weight ratio of 1 to 5; wherein the emulsifier comprises at least one of span 60, span 80, tween 85, sucrose fatty acid ester and lecithin, and the colloid substance comprises at least one of gelatin, gum arabic, guar gum and xanthan gum.
Further, the particle size of the protective agent is less than 80 meshes.
Further, in the step 5), the adding proportion of the protective agent is 0.5% -2% of that of the pretreated yeast paste I.
Further, in the step 3), the mixing weight ratio of the fresh yeast paste to the pretreatment agent solution is 1 to 3:1.
further, in the step 2), the pretreating agent comprises at least one of small molecular saccharides, lower polyols, amino acids and inorganic salts.
Still further, the small molecule saccharide comprises at least one of lactose, galactose, trehalose and isomaltose; the lower polyhydric alcohol comprises at least one of propylene glycol, glycerol and sorbitol; the amino acid comprises at least one of cysteine, alanine and sodium glutamate; the inorganic salt comprises at least one of magnesium sulfate, magnesium chloride, sodium acetate, sodium chloride and dipotassium hydrogen phosphate.
Further, the pretreating agent comprises the following components in a weight ratio of 0.5 to 3:1:1 to 0.5 of mixed small molecule saccharides, amino acids and lower polyols.
Further, filtering in the step 1) and the step 4) by adopting a plate-and-frame filter press.
Compared with the prior art, the invention has the following beneficial effects:
1) The invention prepares the Kluyveromyces marxianus active dry yeast by adopting a two-section fluidized bed drying mode, and combines high-low temperature drying, thereby shortening the drying time, reducing the death of thalli in the drying process, establishing a process which can realize large-scale industrial production, and greatly reducing the production cost compared with a freeze drying process;
2) The heat resistance of the yeast in the drying process is improved by adding the protective agent, and the death of thalli in the drying process is further reduced by combining a two-section drying process, so that the living bacteria retention rate of the Kluyveromyces marxianus in the drying process is improved, the Kluyveromyces marxianus active dry yeast product with high living bacteria number can be prepared, and the product is suitable for the feed industry;
3) According to the invention, the yeast is treated by the pretreatment agent, so that the intracellular environment of the yeast is changed, and the storage resistance of the yeast is improved; meanwhile, the added protective agent has the function of coating and can provide a protective effect on cells; in addition, the lower drying temperature can also reduce the damage of the heat effect on the thallus cells, and the death of the thallus in the storage process is avoided; the combined action of the three aspects improves the storage stability of the Kluyveromyces marxianus active dry yeast product and prolongs the shelf life of the product.
Detailed Description
The technical solutions in the embodiments of the present invention are clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
The embodiment provides a preparation method of kluyveromyces marxianus active dry yeast, which comprises the following steps:
1) Taking 60kg of high-density fermented kluyveromyces marxianus fermentation liquor, filtering by using a plate-and-frame filter press, and collecting fresh yeast paste;
2) Preparing a pretreatment agent into a 5% pretreatment agent solution by using water; the pretreating agent is trehalose;
3) Redissolving the fresh yeast paste obtained in the step 1) in the pretreating agent solution obtained in the step 2), and stirring for 2 hours at 20 ℃ to obtain redissolved yeast suspension; the weight ratio of the fresh yeast paste to the pretreatment agent solution is 1;
4) Filtering the re-dissolved yeast suspension in the step 3) by using a plate-and-frame filter press to obtain pretreated yeast paste I;
5) Scattering the pretreated yeast paste I by using a stirrer, adding a protective agent into the pretreated yeast paste I, wherein the adding proportion of the protective agent is 1% of the weight of the pretreated yeast paste I, and continuously stirring for 20min to obtain pretreated yeast paste II; the protective agent comprises span 60 and gelatin, the weight ratio is 1;
6) Extruding the pretreated yeast paste II into noodle-shaped yeast wet particles by using an extruder, putting the yeast wet particles into a fluidized bed dryer for drying, drying for 15min at 50 ℃, reducing the drying temperature to 37 ℃, and continuously drying for 40min to obtain the Kluyveromyces marxianus active dry yeast product.
Example 2
The embodiment provides a preparation method of kluyveromyces marxianus active dry yeast, which comprises the following steps:
1) 60kg of high-density fermented kluyveromyces marxianus fermentation liquor is taken, filtered by a plate-and-frame filter press, and fresh yeast paste is collected;
2) Preparing a pretreatment agent into a 10% pretreatment agent solution by using water; the pretreatment agent comprises trehalose and cysteine, and the weight ratio is 4;
3) Redissolving the fresh yeast paste obtained in the step 1) in the pretreating agent solution obtained in the step 2), and stirring for 1h at 25 ℃ to obtain redissolved yeast suspension; the weight ratio of the fresh yeast paste to the pretreatment agent solution is 1:1;
4) Filtering the re-dissolved yeast suspension in the step 3) by using a plate-and-frame filter press to obtain pretreated yeast paste I;
5) Scattering the pretreated yeast paste I by using a stirrer, adding a protective agent into the pretreated yeast paste I, wherein the adding proportion of the protective agent is 1.5 percent of the weight of the pretreated yeast paste I, and continuously stirring for 20min to obtain pretreated yeast paste II; the protective agent comprises span 60 and guar gum, the weight ratio is 1;
6) Extruding the pretreated yeast paste II into noodle-shaped yeast wet particles by using an extruder, putting the yeast wet particles into a fluidized bed dryer for drying, drying at 55 ℃ for 10min, reducing the drying temperature to 40 ℃, and continuing to dry for 30min to obtain a Kluyveromyces marxianus active dry yeast product.
Example 3
The embodiment provides a preparation method of kluyveromyces marxianus active dry yeast, which comprises the following steps:
1) 60kg of high-density fermented kluyveromyces marxianus fermentation liquor is taken, filtered by a plate-and-frame filter press, and fresh yeast paste is collected;
2) Preparing a pretreatment agent into a 15% pretreatment agent solution by using water; the pretreatment agent comprises trehalose, alanine and magnesium chloride, and the weight ratio is 1;
3) Redissolving the fresh yeast paste obtained in the step 1) in the pretreating agent solution obtained in the step 2), and stirring for 1.5 hours at 25 ℃ to obtain redissolved yeast suspension; the weight ratio of the fresh yeast paste to the pretreatment agent solution is 2;
4) Filtering the re-dissolved yeast suspension in the step 3) by using a plate-and-frame filter press to obtain pretreated yeast paste I;
5) Scattering the pretreated yeast paste I by using a stirrer, adding a protective agent into the pretreated yeast paste I, wherein the adding proportion of the protective agent is 2% of the weight of the pretreated yeast paste I, and continuously stirring for 20min to obtain pretreated yeast paste II; the protective agent comprises span 60, tween 85, gelatin and gum arabic, the weight ratio of span 60, gelatin and gum arabic is 1;
6) Extruding the pretreated yeast paste II into noodle-shaped yeast wet particles by using an extruder, putting the yeast wet particles into a fluidized bed dryer for drying, drying at 55 ℃ for 15min, reducing the drying temperature to 42 ℃, and continuously drying for 25min to obtain the kluyveromyces marxianus active dry yeast product.
Example 4
The embodiment provides a preparation method of kluyveromyces marxianus active dry yeast, which comprises the following steps:
1) 60kg of high-density fermented Kluyveromyces marxianus fermentation liquor is taken, filtered by a plate-and-frame filter press, and fresh yeast paste kg is collected;
2) Preparing a pretreating agent into a 20% pretreating agent solution by using water; the pretreatment agent comprises trehalose, galactose, alanine, sodium glutamate, glycerol and sorbitol, and the weight ratio is (2);
3) Redissolving the fresh yeast paste obtained in the step 1) in the pretreating agent solution obtained in the step 2), and stirring for 1.5h at 20 ℃ to obtain redissolved yeast suspension; the weight ratio of the fresh yeast paste to the pretreatment agent solution is 3;
4) Filtering the re-dissolved yeast suspension in the step 3) by using a plate-and-frame filter press to obtain pretreated yeast paste I;
5) Scattering the pretreated yeast paste I by using a stirrer, adding a protective agent into the pretreated yeast paste I, wherein the adding proportion of the protective agent is 2% of the weight of the pretreated yeast paste I, and continuously stirring for 30min to obtain pretreated yeast paste II; the protective agent comprises span 60, sucrose fatty acid ester, gelatin and guar gum, wherein the weight ratio of the span 60 to the sucrose fatty acid ester to the gelatin is 2;
6) Extruding the pretreated yeast paste II into noodle-shaped yeast wet particles by using an extruder, putting the yeast wet particles into a fluidized bed dryer for drying, drying for 5min at 65 ℃, reducing the drying temperature to 42 ℃, and continuing to dry for 20min to obtain a kluyveromyces marxianus active dry yeast product.
Example 5
This example provides a method for preparing kluyveromyces marxianus active dry yeast, which is the same as the step of example 4, except that the pretreatment agent in step 2) of this example comprises trehalose and galactose in a weight ratio of 1.
Example 6
This example provides a preparation method of kluyveromyces marxianus active dry yeast, which is the same as the steps of example 4, except that the pretreatment agent in step 2) of this example comprises alanine and sodium glutamate in a weight ratio of 1.
Example 7
The present example provides a preparation method of kluyveromyces marxianus active dry yeast, which is the same as the steps of example 4, except that the pretreatment agent in step 2) of the present example comprises glycerol and sorbitol, and the weight ratio is 1.
Example 8
This example provides a preparation method of kluyveromyces marxianus active dry yeast, which is the same as the steps in example 4, except that the pretreatment agent in step 2) of this example includes trehalose, galactose, glycerol and sorbitol, and the weight ratio is 2.
Example 9
This example provides a method for preparing kluyveromyces marxianus active dry yeast, which is the same as the steps in example 4, except that the pretreatment agent in step 2) of this example comprises alanine, sodium glutamate, glycerol and sorbitol, in a weight ratio of 2.
Example 10
This example provides a method for preparing kluyveromyces marxianus active dry yeast, which is the same as the steps in example 4, except that the pretreatment agent in step 2) of this example comprises trehalose, galactose, alanine, and sodium glutamate in a weight ratio of 1.
Example 11
This example provides a preparation method of kluyveromyces marxianus active dry yeast, which is the same as the steps in example 4, except that the protective agent in step 5) of this example comprises span 60 and sucrose fatty acid ester in a weight ratio of 2.
Example 12
This example provides a preparation method of kluyveromyces marxianus active dry yeast, which is the same as the step of example 4, except that the protective agent in step 5) of this example includes gelatin and guar gum in a weight ratio of 2.
Control test 1
The control test 1 is based on the example 2 and eliminates the step of treating with the pretreatment agent, and the specific test method is as follows:
1) Taking 60kg of high-density fermented kluyveromyces marxianus fermentation liquor, filtering by using a plate-and-frame filter press, and collecting fresh yeast paste;
2) Scattering the fresh yeast paste by using a stirrer, adding a protective agent into the yeast paste, wherein the adding proportion of the protective agent is 1.5 percent of the weight of the fresh yeast paste, and continuously stirring for 20min to obtain pretreated yeast paste II; the protective agent comprises span 60 and guar gum, the weight ratio is 1;
3) Extruding the pretreated yeast paste II into noodle-shaped yeast wet particles by using an extruder, putting the yeast wet particles into a fluidized bed dryer for drying, drying at 55 ℃ for 10min, reducing the drying temperature to 40 ℃, and continuously drying for 30min to obtain the Kluyveromyces marxianus active dry yeast product.
Control experiment 2
The control experiment 2 is a test carried out by changing the drying mode on the basis of the example 2 and adopting the traditional constant-temperature drying mode, and the specific test method is as follows:
1) Taking 60kg of high-density fermented kluyveromyces marxianus fermentation liquor, filtering by using a plate-and-frame filter press, and collecting fresh yeast paste;
2) Preparing a pretreatment agent into a 10% pretreatment agent solution by using water; the pretreatment agent comprises trehalose and cysteine in a weight ratio of 4;
3) Redissolving the fresh yeast paste obtained in the step 1) in the pretreating agent solution obtained in the step 2), and stirring for 1h at 25 ℃ to obtain redissolved yeast suspension; the weight ratio of the fresh yeast paste to the pretreatment agent solution is 1;
4) Filtering the re-dissolved yeast suspension in the step 3) by using a plate-and-frame filter press to obtain pretreated yeast paste I;
5) Scattering the pretreated yeast paste I by using a stirrer, adding a protective agent into the pretreated yeast paste I, wherein the adding proportion of the protective agent is 1.5 percent of the weight of the pretreated yeast paste I, and continuously stirring for 20min to obtain pretreated yeast paste II; the protective agent comprises span 60 and guar gum, the weight ratio is 1;
6) Extruding the pretreated yeast paste II into noodle-shaped yeast wet particles by using an extruder, putting the yeast wet particles into a fluidized bed dryer for drying, and drying at 42 ℃ for 60min to obtain a kluyveromyces marxianus active dry yeast product.
Blank control test
The blank control test is prepared by adopting a common bread yeast preparation process, and the specific method comprises the following steps:
1) 60kg of high-density fermented kluyveromyces marxianus fermentation liquor is taken, filtered by a plate-and-frame filter press, and fresh yeast paste is collected;
2) Scattering the fresh yeast paste by using a stirrer, adding a protective agent into the fresh yeast paste, wherein the adding proportion of the protective agent is 1 percent of the weight of the fresh yeast paste, and continuously stirring for 30min to obtain pretreated yeast paste; the protective agent is span 60, and the grain size of the span 60 is less than 80 meshes;
3) Extruding the pretreated yeast paste into noodle-shaped yeast wet particles by using an extruder, putting the yeast wet particles into a fluidized bed dryer for drying, and drying at 42 ℃ for 60min to obtain a kluyveromyces marxianus active dry yeast product.
The stability evaluation method of the kluyveromyces marxianus active dry yeast comprises the following steps: and (3) taking 20g of dry yeast particles, carrying out vacuum packaging by using an aluminum foil bag, placing the vacuum-packaged sample in an incubator at 37 ℃ for 7 days, taking out the sample, detecting the viable count of the dry yeast particles, calculating the viable survival rate of the dry yeast particles, and evaluating the storage stability according to the viable survival rate.
The viable cell counts of the initial fermentation liquids, the initial viable cell counts of the active dry yeasts and the viable cell counts of the dry yeast samples after being stored for 7 days at 37 ℃ in the examples 1 to 12, the blank control test and the control test 1-2 were respectively measured, and the viable cell retention rate of the accelerated storage in the drying process corresponding to each example and the control test were calculated, and the results are shown in table 1.
TABLE 1 results of measurements relating to Kluyveromyces marxianus prepared in examples 1-12 and the control experiment
As can be seen from Table 1, the viable count of the Kluyveromyces marxianus active dry yeast product prepared by the preparation method can reach 2 to 4 multiplied by 10 10 CFU/g, suitable for addition to animal feed.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and should not be taken as limiting the scope of the present invention, which is intended to cover any modifications, equivalents, improvements, etc. within the spirit and scope of the present invention.
Claims (7)
1. A preparation method of kluyveromyces marxianus active dry yeast is characterized by comprising the following steps:
1) Filtering kluyveromyces marxianus fermentation liquor obtained by high-density fermentation to obtain fresh yeast paste;
2) Preparing a pretreating agent into a pretreating agent solution with the concentration of 5-20% by using water; the pretreating agent comprises the following components in a weight ratio of 0.5-3: 1: 1-0.5 mixed small molecular saccharides, amino acids and lower polyols;
3) Redissolving the fresh yeast paste obtained in the step 1) in the pretreating agent solution obtained in the step 2), and stirring for 0.5-2 h at the temperature of 20-25 ℃ to obtain redissolved yeast suspension;
4) Filtering the re-dissolved yeast suspension in the step 3) to obtain pretreated yeast paste I;
5) Scattering the pretreated yeast paste I by using a stirrer, adding a protective agent into the yeast paste I, and continuously stirring for 20-30 min to obtain pretreated yeast paste II; the protective agent comprises an emulsifier and a colloid substance which are mixed according to the weight ratio of 1;
6) Extruding the pretreated yeast paste II into noodle-shaped yeast wet particles by using an extruder, then putting the yeast wet particles into a fluidized bed dryer for drying in two sections, drying at 50-65 ℃ for 5-15 min in the first stage, controlling the moisture of the material to be 25% -35% when the drying is finished, reducing the drying temperature to 35-42 ℃ in the second stage until the moisture of the material is 4% -6%, and finishing the drying to obtain the Kluyveromyces marxianus active dry yeast product.
2. The method for producing kluyveromyces marxianus active dry yeast as claimed in claim 1, wherein: in the step 5), the emulsifier comprises at least one of span 60, span 80, tween 85, sucrose fatty acid ester and lecithin, and the colloidal substance comprises at least one of gelatin, gum arabic, guar gum and xanthan gum.
3. The method for producing kluyveromyces marxianus active dry yeast as claimed in claim 2, wherein: the particle size of the protective agent is less than 80 meshes.
4. The method for preparing kluyveromyces marxianus active dry yeast as claimed in claim 1, wherein: in the step 5), the adding proportion of the protective agent is 0.5-2% of that of the pretreated yeast paste I.
5. The method for preparing kluyveromyces marxianus active dry yeast as claimed in claim 1, wherein:
in the step 3), the mixing weight ratio of the fresh yeast paste to the pretreatment agent solution is 1-3: 1.
6. the method for preparing kluyveromyces marxianus active dry yeast as claimed in claim 1, wherein: the small molecular saccharides comprise at least one of lactose, galactose, trehalose and isomaltose; the lower polyhydric alcohol comprises at least one of propylene glycol, glycerol and sorbitol; the amino acid comprises at least one of cysteine, alanine and sodium glutamate.
7. The method for preparing kluyveromyces marxianus active dry yeast as claimed in claim 1, wherein:
filtering in the step 1) and the step 4) by adopting a plate-and-frame filter press.
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