JP2001309751A - Additive for feed - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an additive for a feed consisting essentially of an amino acid good in fluidity by carrying out an amino acid fermentation using a crude saccharified liquid raw material available at a low cost and directly drying and granulating the resultant fermentation broth. SOLUTION: This additive for the feed is characterized by suspending a cassava tuber peeled, dried and pulverized to <=150 μm in water saccharifying the resultant suspension of the obtained powder, providing a saccharified liquid containing solid residues such as fibrous substances, carrying out an amino acid fermentation using the saccharide liquid as a fermentation raw material and drying and granulating the resultant amino acid fermentation broth.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a method for producing a sugar solution containing solid residues such as fibers by directly saccharifying tapioca starch from cassava potato tissue without separating and purifying the same, and using the sugar solution as a raw material for fermentation. The present invention relates to an additive for feed containing an amino acid as a main component, which is obtained by subjecting an amino acid fermented solution obtained by subjecting an obtained amino acid fermentation solution to dry granulation.

[0002]

2. Description of the Related Art When amino acids are used as feed additives, they do not necessarily need to be of high purity.
Many attempts have been made to produce a feed additive containing an amino acid as a main component by drying directly from a fermentation broth containing an amino acid obtained by fermentation. However, the powder thus obtained has remarkable hygroscopicity due to the influence of many impurities such as saccharides and organic acids contained in the broth, and the powder is often condensed. It often becomes a large lump, and its handling is often poor.

[0003] In order to prevent the caking due to moisture absorption and to produce a powder having good handleability, a method is used in which a specific additive is mixed with a fermentation broth of an amino acid and then dried. For example, in Japanese Patent Publication No. 43-22235, L
-A method of concentrating a fermentation broth containing lysine hydrochloride, mixing it with bran, which is a feed material, and drying it. Further, in French Patent 2,217,347, a dehydrating agent such as powdered silica, expanded perlite, bone powder, bran, calcium carbonate or fluorine-free phosphate is mixed with the fermentation broth and dried. Furthermore, Japanese Patent Application Laid-Open No. 5-192089 discloses that granules having a low protein content and excellent handleability as compared with conventional products are obtained by removing biomass such as cells from the amino acid fermentation broth, concentrating and drying. It is described that can be obtained.

Originally, feed additives had to be inexpensive, and thus were inexpensive without adding a bran or a dehydrating agent to the amino acid fermentation broth or without passing through a step of removing cells from the amino acid fermentation broth. It is extremely significant if an amino acid can be fermentatively produced from various fermentation raw materials, and the fermentation broth can be directly or partially concentrated and then subjected to dry granulation to produce granules having good fluidity.

[0005] In the fermentation method of amino acids, if an insoluble solid is present in the culture solution after fermentation, the operation of isolating and purifying the target amino acid is hindered. It has been.

[0006] Tapioca is a starch produced from cassava potato, converted into glucose by enzymatic saccharification, and used as a raw material for amino acid fermentation.

[0007] Conventionally, tapioca has been prepared by grinding cassava potato as a raw material, extracting starch fine particles stored in the potato with water, and straining it with a fine mesh or centrifuging to obtain a fiber other than starch. It is manufactured by separating and collecting from scum and drying. Usually, the raw cassava potato (raw potato) contains about 50% water,
At present, tapioca is obtained at a yield of 0 to 25%, and almost the same amount of scum is discharged.

[0008]

However, when the starch content of cassava potato was analyzed, 40-45% of the raw potato was starch, and therefore, a considerable amount of starch was not extracted, and the discharged fiber Etc. will remain in the residue.

Cassava potatoes are usually crushed by a sawtooth tooth implantable grinder (raspar) or the like. However, since the cell membrane is thicker and the starch particles are smaller than potatoes, the crushing efficiency is slightly lower. It is not sufficient for separation and no remarkable improvement in starch yield is observed. In fact, when the scum is observed with a scanning electron microscope, it can be clearly seen that almost homogeneous and spherical starch granules are present and many starch particles are covered with a film-like material.

[0010] In order to reproduce the same state as the crushing of cassava potato in a laboratory, cassava potatoes were grated with a grater according to a flow sheet shown in FIG. 1 and a starch production experiment was carried out. Each weight of starch and scum was measured to determine the starch yield (extraction rate). Table 1 shows the results of three experiments performed under the same conditions.

[0011]

[Table 1]

[0012] As shown in Table 1, the weight of the starch and the weight of the scum obtained in each case were almost the same, and almost the same amount of scum as the product tapioca was discharged at the actual tapioca starch factory. Well matched with the situation.

[0013] As described above, many starch particles remain in the scum and are covered with a film-like material, so that the usual crushing process for collecting starch similarly imposes a great restriction on the yield of starch. As a result, the starch in the cassava potato cannot be sufficiently extracted. This is not only because the starch resources in cassava potatoes are not being used effectively, but also because there is still a large amount of starch in waste such as scum discharged from starch manufacturing plants. May be connected.

Therefore, a high-concentration saccharified solution is produced by directly saccharifying a cassava potato without passing through a starch extraction step,
Using this as a fermentation raw material to produce amino acids by fermentation, and by directly drying and granulating the fermentation broth to produce a feed additive containing amino acids as a main component that has substantially no tackiness, it can be produced from cassava potatoes. Not only is it possible to significantly reduce scum and the like generated during the production of starch, but also to make effective use of it.

The present invention has been made in view of such a viewpoint, and tapioca starch is directly saccharified from cassava potato without separation / purification to produce a high-concentration saccharified solution, and this high-concentration saccharified solution is used. The purpose of the present invention is to produce an amino acid-based feed additive by fermenting and producing an amino acid and then drying and granulating the fermentation broth.

[0016]

Means for Solving the Problems The present inventors have made intensive studies to achieve the above object, and as a result, peeled, dried and finely ground cassava potatoes destroyed the film covering the starch particles by this finely pulverizing process. It is possible to saccharify the starch in the saccharified solution, and this saccharified solution is suitable as a raw material for fermentation of amino acids, the amino acids can be obtained in high yield, and the utilization rate of the starch of cassava potato can be significantly increased. Concentration of fermentation broth of amino acids containing solid residues such as fibrous and bacterial cells derived from cassava potatoes, followed by dry granulation, yielding feed additives mainly composed of amino acids with good fluidity. Was found.

The present invention has been made based on this finding. Cassava potato, which has been peeled, dried, and crushed to 150 μm or less, is suspended in water to saccharify the sugar solution containing a solid residue such as fiber. The present invention relates to a feed additive containing an amino acid as a main component, characterized in that an amino acid is fermented and produced by using this as a fermentation raw material, and the obtained amino acid fermented liquid is dried and granulated.

[0018]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail.

The raw cassava potato used as a raw material is
Since the quality is greatly reduced after harvesting, it can be preserved by immediately peeling and drying, and the peeled and dried potato can be made into a fine powder similar to tapioca starch by pulverizing with a crusher. The starch content per dry matter is about 90%, and thus it is a starch-based crude starch powder.

The cassava potato having a water content of 16% by weight or less is dried for 1 to 10 mm using a crusher or the like according to a conventional method.
Crushed into particles. Subsequently, the material is pulverized to a size of 150 μm or less that passes through a 100 mesh. In the case of mass processing on an industrial scale, examples of a device for performing pulverization include a pin mill and a ball mill.

The smaller the water content of the powder, the higher the weight ratio of the powder having a size of 150 μm or less, and the easier it is to liquefy and saccharify. Therefore, the water content is 16% by weight or less, preferably 5 to 10% by weight. Dry powder is used.

The pulverized powder having a particle size of 150 μm or less has good suspension properties, maintains good fluidity even at a slurry concentration of 45%, and has a low viscosity of less than 500 centipoise and good operability. By making the particles finer, advantages such as a shorter liquefaction time and easier handling of the liquid can be enjoyed.

The powdered cassava potato can be directly liquefied and saccharified by using a starch liquefaction enzyme and a saccharification enzyme like tapioca starch. When the powder of the dried potato is liquefied and saccharified, the starch becomes glucose and is solubilized, but components other than starch, such as fibrin, remain as solids. After the completion of saccharification, the saccharified solution containing solid matter such as cellulose is used as it is as a raw material for amino acid fermentation in the next step.

When the dried cassava potato powder is suspended in water, if it is subjected to an enzymatic treatment with cellulase before liquefaction, it can be suspended at a high concentration, promoting liquefaction and saccharification, and reducing the saccharification rate. improves.

When the whole amount of the dried cassava potato powder to be liquefied in the cellulase-containing liquid is not added all at once but is added in small portions, the suspension can be suspended at a high concentration of 45 to 55 g / dl. Can be produced.

By the action of cellulase, the membrane covering the starch particles is enzymatically degraded, resulting in liquefaction of the starch.
It is considered that saccharification is effectively performed. As the cellulase, a commercially available cellulase such as “CELLULASE YC” (trade name, manufactured by Kikkoman Corporation) can be used, and a cellulase produced by a fungus of the genus Trichoderma is preferably used. The amount to be used is 10 with filter paper disintegration activity (FPA) (see Jinjinshokan “Enzyme Utilization Handbook”, pages 297 to 298).
What is necessary is just about a unit or more. 150 per gram of powder
The unit is at least a unit, preferably 300 to 1500 units. The reaction is carried out in a temperature range of 40-50 ° C, pH 5-7. The reaction time may be generally about 2 to 4 hours.

After performing the cellulase reaction, a liquefying enzyme, for example, “T-5” (trade name, manufactured by Daiwa Kasei Co., Ltd.) is added to powder 1
85 to 95 units
Stir at 1 ° C. for 1 hour. After allowing to cool to room temperature and adjusting the pH to 4.2 with dilute sulfuric acid, the saccharifying enzyme, for example, "NC-4.2" [trade name, Amano Pharmaceutical Co., Ltd.]
Was added so as to be 2 to 5 units per 1 g of powder, heated to 55 to 65 ° C, and kept at 42 to
Heat and stir for 48 hours to complete the enzyme reaction.

When suspending the dried cassava potato powder in water, liquefaction proceeds more quickly if 20 units / g of powdered liquefaction enzyme are added in advance. Also, without heating to 90 ° C., where liquefaction enzyme activity is high, liquefaction / saccharification efficiency is increased by gradually adding powder to a liquefaction enzyme-containing solution at 40 to 60 ° C., and high concentration (45 to 55 g / dl) is obtained. The powder can be saccharified to produce a sugar solution having a glucose concentration of 40 to 45 g / dl.

The relationship between the particle size of the dried peeled potato powder, the saccharification ratio and the residue ratio is shown in Experimental Example 1 below.

Experimental Example 1 Immediately after harvesting a potato from Lampung, Indonesia, the skin was peeled off, dried in the sun until the weight was reduced to 2/3, and further dried for 40 days.
The mixture was dried under reduced pressure at about 24 ° C. for about 24 hours until no change in weight was observed. The water content of this powder was about 5%. The powder was acid-decomposed (7.6N HCl was added and heated in boiling water for 2 hours), and the starch content as glucose was determined by HPLC. The starch content in the powder was 89.4%.
Met.

A dried potato of about 10 to 20 cm in length is 1 cm.
Crushed to ² and the desktop blender "Osterizer"
16-speed "[SUN BEAM OSTER
Manufactured by the company).

The pulverized powder is mixed with 9, 16, 40, 100
The powder was classified into four types that passed through the mesh, and 5 g of powder of each particle size was stirred in advance with a liquid to which liquefaction enzyme “T-5” (manufactured by Daiwa Kasei Co., Ltd.) (20 units / g of powder) was added. While adding in 5 portions. After completion of the addition, the mixture was further reacted for 1 hour in boiling water while shaking and stirring. The mixture was allowed to cool to room temperature, and the pH was adjusted to 4.2 with dilute sulfuric acid. The saccharification enzyme "NC-4.2" is added in an amount of 3
The unit was added, heated to 60 ° C., and stirred at the same temperature for 48 hours. The liquid after completion of saccharification was centrifuged, and the saccharification rate and residue ratio of the saccharified liquid were analyzed.

As shown in Table 1, the relationship between the particle size and the saccharification ratio and residue ratio is shown in Table 1. As the particle size becomes smaller, the saccharification is smoother. 82% glucose was obtained per powder, indicating that 94% of the starch contained was saccharified.

[0034]

[Table 2]

Next, the moisture content of the dried peeled potatoes and a 100 mesh pass (%) when crushed for a certain time using a crusher.
Is shown in Experimental Example 2.

Experimental Example 2 Desktop blender "Osterizer 16-spe"
45 g of peeled potatoes having a size of about 3 to 5 cm each adjusted to a water content of 8 to 22% in a constant humidity container using "ed" were crushed for 4 minutes.

The pulverized sample was dried at 105 ° C. for 4 hours, sieved with 100 mesh, and the weight% of the sample that passed through 100 mesh was calculated. The relationship between the water content (%) of the peeled and dried potatoes and the 100 mesh pass (%) is as shown in FIG. 2, when the sample water content exceeds 16%.
The ratio that can be pulverized to the target particle size of 150 μm or less greatly decreases, and if it exceeds 17%, the ratio that does not reach the target particle size is 1
The sample moisture content is increased by 16% because it increases by 0% or more.
When the pulverization is performed in the following manner, the pulverizable ratio reaches 78% (about 35 g / batch) up to 150 μm or less, and the pulverization efficiency is good, and a high-concentration slurry can be charged.

Experimental Example 3 shows the enzymatic treatment effect when cellulase treatment was performed before liquefaction when the dried cassava potato powder was suspended in water.

EXPERIMENTAL EXAMPLE 3 A solution prepared by dissolving 0.3 g of cellulase "CELLULASE YC" in 7.2 ml of water was used in Example 1.
5 g of cassava potato powder (water content: about 5%) of a 00 mesh pass was added little by little while stirring. After the total amount was added, the mixture was reacted at 50 ° C. for 2 hours while further shaking and stirring. After adding 7 ml of water, the liquefaction enzyme “T-
5 "(0.02 ml, 20 units / g of powder) was added, and the mixture was shaken and stirred at 95 ° C for 1 hour. The mixture was allowed to cool to room temperature, the pH was adjusted to 4.2 with dilute sulfuric acid, 0.004 ml (3 units / g of powder) of saccharifying enzyme “NC-4.2” was added, and the mixture was heated to 60 ° C. The mixture was stirred at the same temperature for 48 hours. The liquid after completion of saccharification was centrifuged, and the saccharification rate and residue ratio of the saccharified liquid were analyzed. For comparison, Table 3 also shows the results obtained when the same operation was performed without using cellulase.

[0040]

[Table 3]

From the results shown in Table 3, it can be seen that the cellulase treatment before the liquefaction has a higher saccharification rate and a smaller residue rate than the cellulase untreated section. Therefore, it can be said that the cellulase treatment of the powdered cassava is effective in improving the efficiency and the quality of the sugar solution in the saccharification performed thereafter.

Even if the cellulase has a concentration of about 10 FPA (unit / ml), by adding the powder little by little, the fluidity can be maintained up to a high concentration (50 g / dl). Liquefaction / saccharification is smooth and gives a high concentration of sugar solution.

Experimental Example 4 shows the effect of the liquefied enzyme when the liquefied enzyme was added in advance when the dried cassava potato powder was suspended in water.

Experimental Example 4 A liquid obtained by adding a predetermined amount of liquefying enzyme "T-5" to 4 ml of water was kept at 40 ° C., and 6 g of powdered cassava potato (with a water content of about 5%) of 100 mesh pass was gradually added thereto. The total volume was 10 ml, and the slurry was liquefied and saccharified under the same conditions as in Experimental Example 1. The amount of liquefied enzyme to be added is set to 100 units / g, which is a standard amount with respect to the amount of starch, a double amount thereof, and no addition. The enzyme was added.

At the time of powder suspension, there was no difference in the ease of suspension depending on the amount of the enzyme. However, when the temperature was increased and then liquefaction was carried out, the liquefaction proceeded more quickly if the enzyme had been added in advance. The liquid became completely smooth in 30 minutes, which is half of the liquefaction reaction time. At that time, the sample to which the enzyme had not been previously added was not liquefied. Table 4 shows the residue ratio and saccharification ratio when each enzyme amount was added.

[0046]

[Table 4]

From the results shown in Table 4, it can be seen that there is a significant difference in the residue ratio and the saccharification ratio in the group where the enzyme was added in advance compared to the group where no enzyme was added, indicating that liquefaction and saccharification can be performed efficiently.

Also, 55 g of powdered cassava potato is gradually added to make the total volume 100 ml. The temperature set at the time of powder suspension is 40 ° C. before liquefaction, 60 ° C. immediately before gelatinization, and 80 ° C. having high liquefying enzyme activity. Liquefaction and saccharification were performed under the same conditions as in Experimental Example 1 except that the temperature was changed to ° C. Table 5 shows the results.

[0049]

[Table 5]

There is no great difference in the concentration of sugar solution. In comparison with the temperature at the time of suspension under the same conditions, the result of the best saccharification rate was 60 ° C, but the smoothness of the suspension was the best at 40 ° C. At 80 ° C., an increase in viscosity was observed, and there was much adhesion to the wall surface of the reaction vessel.

In the present invention, the sugar solution of the dried cassava potato thus obtained is used as a raw material for amino acid fermentation.

Examples of the type of amino acid fermentation include fermentation of amino acids such as lysine, tryptophan, and threonine.

The microorganisms used for these fermentations may be those known for each fermentation. An example is as follows. L-threonine fermenting bacteria: Brevibacterium flavum ATCC 21269 L-tryptophan fermenting bacteria: Bacillus subtilis FERM BP-208 L-lysine fermenting bacteria: Brevibacterium lactofermentum ATCC 21800

The medium used for fermentation may be the same as a known medium except that the sugar solution of the present invention is used as a carbon source. The above sugar solution may be used for the entire amount of the carbon source, or may be used for a part thereof. The remaining carbon source when used for one part may be the same as that used conventionally.

The fermentation method may be the same as the conventional method. However, when the sugar solution of cassava potato is used, the growth rate of the bacteria is faster than in the conventional method using glucose as a fermentation raw material, and the amount of cells increases by about 20% after the fermentation is completed. I do. The fermentation time can be reduced by about 1 to 5 hours.

The concentration of each amino acid obtained by fermentation and the yield to sugar are also almost the same as those in the past.

The fermented broth contains 10 to 30% by weight of cassava potato-derived scum and bacteria derived from cassava potato as it is saccharified and used. The total solids content in the fermentation liquor before concentration is generally between 15 and 25% by weight.
The fermented liquor is dried and granulated as it is or concentrated until the total solid content is about 20 to 50% by weight, and the concentrated liquid is dried and granulated to have an amino acid content of 30 to 80 in terms of dry matter.
% By weight, preferably 60 to 75% by weight of the feed additive.

As a method for granulating and drying the fermented solution of amino acids, there are a method of first solidifying the fermented solution and granulating the solidified product, and a method of directly granulating and drying the amino acid-containing solution using seed crystals. be able to. First, in the former method, as a method for solidifying a fermentation solution of an amino acid, a drying and solidifying method using a spray drier, a drum drier, a spin flash drier or the like can be applied. When the solidified product of the obtained amino acid fermentation solution is fine particles, a method of granulating the solidified product using various granulators such as a granulation mixer, a fluidized bed granulator, and a press granulator is possible. If a fermentation broth itself or an amino acid-containing liquid such as a liquid from which impurities such as bacterial cells in the fermentation broth are separated and removed is used as a binder for the obtained fine particles, granulation can be easily performed. In the case of a solidified gab flake-like dried product of the obtained amino acid-containing liquid, the above granulation method can be applied by pulverizing with a pin mill or the like. In the fluidized-bed granulation, heavy granules can be easily obtained by applying the rolling-type fluidized-bed granulation, and it is easy to obtain a granulated product suitable for the present invention. In the pressure granulation, the solidified product of the amino acid-containing liquid can be applied as it is without pulverization, even if it is in the form of flakes. In addition, depending on the composition of the amino acid-containing liquid such as fermentation broth, the dried product may have tackiness, but this property has an effective effect on granulation, particularly when using pressure granulation.

On the other hand, as a method of directly granulating and drying the amino acid fermentation liquid using the seed crystal, a solidified material of the amino acid fermentation liquid or the above-described amino acid-containing powder is used as a seed crystal, and the fluidized bed granulation is performed. The method of spraying the amino acid-containing liquid into the layer by applying the above method and directly drying and granulating the fermented liquid of the amino acid can be applied. As the seed crystal, the amino acid-containing powder described above can be used. For example, a part of a product obtained by fluidized bed granulation can be pulverized and used as a seed crystal.
In the fluidized-bed granulation, especially by applying the rolling-type fluidized-bed granulation, it is easy to obtain a product having a large bulk density, and it is easy to obtain a granulated product suitable for application to the present invention.

The feed additive thus obtained is
Because of its good fluidity and low consolidation and cohesion, it is a granule with good mixing properties and easy handling.

[0061]

EXAMPLES Sugar solution production example 1 Cassava potato powder (moisture content 5%) of 100 mesh pass
About 450 g of water was suspended in water and adjusted to a total volume of 1 L. This was heated to 85 ° C. with stirring, 1.5 ml of liquefied enzyme “T-5” was added, and a liquefaction reaction was performed for 1 hour. After the completion of liquefaction, when the temperature was cooled to 60 ° C., the pH was adjusted to 4.5 with dilute sulfuric acid, and 0.5 ml of saccharifying enzyme “NC-4.2” was added. The saccharification reaction was performed at 60 ° C. with shaking and stirring for 48 hours. The glucose concentration in the sugar solution is 3
It was 7.5 g / dl (saccharification rate 90%).

Comparative Example of Sugar Solution Production Liquefaction / saccharification was performed in the same manner as in Sugar Solution Production Example 1, except that dried cassava potato powder having a water content of 20% (100 mesh pass ratio: 52%) was used. Where
The glucose concentration in the sugar solution was 20.8 g / dl (saccharification rate 5
0%).

Sugar Liquid Production Example 2 40 g of cellulase "CELLULASE YC" was added to water 1
The solution, which was stirred and dissolved in 0 L, was heated to 40 ° C., and stirred with a 100-mesh pass cassava potato powder (starch content 89%).
%) Was added little by little over 1 hour. After adding the whole amount of cassava potato powder, the mixture was reacted at 50 ° C. for 2 hours while stirring.

33.3 ml of the liquefying enzyme "T-5" was added, and the mixture was shaken and stirred at 90 ° C. for 1 hour. The mixture was allowed to cool to 60 ° C., the pH was adjusted to 4.2 with dilute sulfuric acid, 11.3 ml of saccharifying enzyme “NC-4.2” was added, heated to 60 ° C., and stirred at the same temperature for 48 hours. did. After cooling to room temperature, 18.5 L of a sugar solution was obtained. The glucose concentration in this sugar solution is 44
g / dl (saccharification rate 87%).

Sugar Liquid Production Example 3 A solution obtained by stirring and dissolving 133 ml of the liquefying enzyme "T-5" in 20 L of water was heated to 40 ° C., and 17 kg of cassava potato powder (starch content: 74%) with a 100 mesh pass was stirred under stirring. 1
It was added in small portions over time. Stirring was performed at 90 ° C. for 1 hour. Cool to room temperature and adjust the pH to 4.2 with dilute sulfuric acid.
, And 20 ml of saccharifying enzyme "NC-4.2" was added, heated to 60 ° C, and stirred at the same temperature for 48 hours. After cooling to room temperature, 37.5 L of a sugar solution was obtained. The glucose concentration in this sugar solution was 35 g / dl (saccharification rate 97%).

Fermentation Example 1 Glucose 30 g / L, ammonium chloride 10 g / L,
Urea 3 g / L, KH ₂ PO ₄ 1 g / L, MgSO _{4.
7H 2 O 100mg / L, FeSO} 4 · 7H 2 O 10
mg / L, 8 mg / L of MnSO ₄ .4H ₂ O, 1 g / L of soybean protein hydrolyzate (as nitrogen), 0.1 mg / L of thiamine hydrochloride and 0.3 mg / L of biotin (pH 7.0) ) Was dispensed into three 500 ml shake flasks, each 20 mL. After heat sterilization at 115 ° C. for 10 minutes, the medium was preliminarily added to
31. A platinum loop of Brevibacterium lactofermentum ATCC 21800 grown for 8 hours was inoculated.
Shaking culture was performed at 5 ° C. for 24 hours. The above is a seed culture.
80 g / L as molasses, 50 g / ammonium sulfate
_{L, KH 2 PO 4 1g /} L, MgSO 4 · 7H 2 O 1
g / L, soy protein hydrolyzate (as nitrogen) 100m
g / L, 0.1 mg / L of thiamine hydrochloride, and 0.3 mg / L of biotin (pH 7.0)
Each 300 mL was dispensed into a small 1 L glass fermenter and sterilized by heating at 120 ° C. for 15 minutes. After cooling to 31.5 ° C., the above-mentioned flask culture ending solution was added to the fermenter for 15 times.
Add 3 mL at a time, temperature 31.5 ° C, aeration rate 1 / 2vv
The culture was performed under the conditions of m and the number of agitation 700 rpm. Feeding of the feed medium was started when the sugar concentration in the culture solution became 5 g / L or less. The medium was glucose (40 g /
dL) or a sugar solution obtained by adding glucose to the sugar solution obtained in the sugar solution production example 3 to adjust the glucose concentration to 40 g / dL, a soybean protein hydrolyzate (100 m2 as nitrogen).
g / L), thiamine hydrochloride (0.1 mg / L) and biotin (0.3 mg / L). In each case, the culture was continued while adjusting the feed rate of the feed medium so that the sugar concentration in the culture solution was 5 g / L or less.
After the feed of 00 mL of the feed medium was completed, the culture was terminated when the sugar in the culture was consumed, and the concentration of L-lysine accumulated in the culture was quantified. As a result of the culture of the two plants, the yield in the case of the glucose feed was 35%, while the yield in the case of the cassava feed was 38%.

Fermentation Example 2 The sugar solution obtained in the sugar solution production example 3 was used to prepare L in a 50 L jar.
-A lysine fermentation test was performed. For fermentation, the seed culture liquid volume was 2.75 l, the liquid volume at the start of fermentation was 18 l, and stirring was 250
The fermentation was carried out under the same conditions as in Fermentation Example 1 except that the rpm and the aeration were ｖ vvm. As a result of the fermentation, the yield in the case of the glucose feed was 34%, while the yield in the case of the cassava feed was 37%.

Fermentation Example 3 13.0 g / dL glucose, 4 g / ammonium sulfate
_{dL, KH 2 PO 4 0.7g /} dL, MgSO 4 · 7H
_{2 O 0.04mg / dL, FeSO 4} · 7H 2 O 1m
g / dL, MnSO ₄ .4H ₂ O 1 mg / dL, thiamine hydrochloride 200 μg / dL, casamino acid 0.1 g / dL
dL, soy protein acid hydrolyzate (as total nitrogen) 30 mg
/ DL, 20 mg / dL of L-phenylalanine, 300 μg / dL of biotin, and 100 mg / dL of waste effluent (as total nitrogen) at 115 ° C. for 10 hours.
Heat sterilization for minutes. To this was added potassium carbonate pre-sterilized by dry heat at a rate of 5 g / dL. In this medium, Bacillus subtilis FERM BP-20 cultured in advance was added.
One loopful of 8 was inoculated and shake-cultured at 30 ° C. until glucose disappeared.

Further, instead of using glucose alone as a sugar raw material, the sugar solution obtained in Sugar Solution Production Example 3 was converted to glucose by 1%.
Culture was carried out as above using an amount corresponding to 3.0 g / dL. Sugar solution production example 3 when glucose alone was used
Also when the sugar solution obtained in was used, the yield of L-tryptophan relative to sugar was 5%.

Fermentation Example 4 Glucose 3 g / dL, ammonium sulfate 0.2 g / d
L, urea 0.2 g / dL, KH ₂ PO ₄ 0.15 g / d
L, MgSO ₄ .7H ₂ O 0.04 mg / dL, thiamine hydrochloride 100 μg / dL, biotin 300 μg / d
dL and soybean protein acid hydrolyzate, 140m as total nitrogen
The medium containing g / dL was adjusted to pH 7.0 and
1 at a temperature of 115 ° C. in a 500 ml shoulder flask.
Heat sterilization for 0 minutes. One platinum loop of Brevibacterium flavum ATCC 21269 was inoculated into this medium in advance, and cultured with shaking at 31.5 ° C. for 12 hours. The above is the seed culture. In the main culture, glucose 13 g / d
_{L, KH 2 PO 4 0.25g /} dL, MgSO 4 · 7
H ₂ O 40 mg / dL, ammonium sulfate 2.0 g /
_{dL, FeSO 4 · 7H 2 O} 1mg / dL, L- isoleucine 40 mg / dL, biotin 50 [mu] g / dL, thiamine hydrochloride 5 [mu] g / dL, soybean protein acid hydrolyzate (as total nitrogen) 32 mg / dL and Suteffen effluent (total 20 ml of medium adjusted to pH 6.5 with 200 mg / dL (as nitrogen) was dispensed into a 500 ml shoulder flask and steam sterilized at 110 ° C. for 10 minutes. To this was added 2 g of potassium carbonate which had been dry-heat sterilized in advance, and added to 1 ml of the above seed culture. Culture was performed at a temperature of 31.5 ° C. In the main culture, 13 g / d of glucose was used as a sugar material.
Culture was performed under the same conditions except that the sugar solution obtained in Sugar Solution Production Example 3 was used in place of L in an amount corresponding to 13 g / dL in terms of glucose. When the sugar in the culture solution was consumed, the culture was terminated, and the concentration of L-threonine accumulated in the culture solution was quantified. L- when only glucose is used as a sugar raw material
The yield with respect to sugar of threonine was 12.0%, whereas the yield using cassava sugar solution was 11.8%.
Met.

Production Example 1 of Feed Additives The fermented liquors obtained in Fermentation Examples 1 and 2 were concentrated as they were or until the solid content became 40%. MINI ”) under the conditions shown in Table 6. The physical properties of the obtained granules and the results of caking / moisture absorption tests are shown in the same table.

[0072]

[Table 6]

Consolidation / Hygroscopicity Test: About 5 g of a sample was placed in a styrene bottle and the consolidation was evaluated when stored for 8 days in a 20 ° C. constant humidity bath with a saturated solution of magnesium chloride (relative humidity: 33%). did. The relative humidity is 33%,
Moisture absorption (%) when stored at 3%, 65%, 76%
Is shown in FIG.

Criteria for evaluation of solidification: see [Table 7]

[0075]

[Table 7]

The granulated product had an L-lysine content of about 63% and a protein content of about 11%, and it was confirmed that the granulated product could be used as a good feed additive having a higher L-lysine content than conventional ones.

Preparation Example 2 for Feed Additives The fermented L-threonine solution obtained in Fermentation Example 4 was subjected to spray granulation in the same manner as in Preparation Example 1 for feed additives to obtain granules containing L-threonine.

[0078]

As described above, according to the present invention, powdered cassava potato is directly saccharified, amino acid fermentation is carried out using the raw material as a raw material, and the obtained amino acid fermented liquid is dried and granulated, so that the powder is less powdery and can be handled. It is possible to provide an inexpensive feed additive having a high amino acid content easily.

[Brief description of the drawings]

FIG. 1 is a flow sheet showing an example of producing starch and scum from cassava potatoes.

FIG. 2 is a graph showing the relationship between the water content (%) of peeled and dried cassava potatoes and the weight ratio (%) of powder having a particle size of 150 μm or less.

FIG. 3 is a graph showing the change over time of the moisture absorption of the L-lysine-containing feed additive according to the present invention at 20 ° C. and a relative humidity of 33%, 43%, 65%, and 76%.

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[Procedure amendment]

[Submission date] May 11, 2001 (2001.5.1
1)

[Procedure amendment 1]

[Document name to be amended] Statement

[Correction target item name] 0033

[Correction method] Change

[Correction contents]

As a result of examining the relationship between the particle size and the saccharification ratio and residue ratio, as shown in Table 2, the finer the particle size, the smoother the saccharification. By pulverizing to a particle size of 150 μm or less passing through 100 mesh. 82% glucose was obtained per powder, indicating that 94% of the starch contained was saccharified.

[Procedure amendment 2]

[Document name to be amended] Statement

[Correction target item name] 0058

[Correction method] Change

[Correction contents]

As a method of drying and granulating the fermented solution of amino acid, there are a method of first solidifying the fermented solution and granulating the solidified product, and a method of directly drying and granulating the amino acid-containing solution using seed crystals. be able to. First, in the former method, as a method for solidifying a fermentation solution of an amino acid, a drying and solidifying method using a spray drier, a drum drier, a spin flash drier or the like can be applied. When the solidified product of the obtained amino acid fermentation solution is fine particles, a method of granulating the solidified product using various granulators such as a granulation mixer, a fluidized bed granulator, and a press granulator is possible. If a fermentation broth itself or an amino acid-containing liquid such as a liquid from which impurities such as bacterial cells in the fermentation broth are separated and removed is used as a binder for the obtained fine particles, granulation can be easily performed. When the solidified product of the obtained amino acid-containing liquid is a flake-form dried product, the above granulation method can be applied by pulverizing with a pin mill or the like. In the fluidized-bed granulation, heavy granules can be easily obtained by applying the rolling-type fluidized-bed granulation, and it is easy to obtain a granulated product suitable for the present invention. In pressure granulation, even when the solidified product of the amino acid-containing liquid is in the form of flakes, it can be applied as it is without pulverization. In addition, depending on the composition of the amino acid-containing liquid such as fermentation broth, the dried product may have tackiness, but this property has an effective effect on granulation, particularly when using pressure granulation.

[Procedure amendment 3]

[Document name to be amended] Statement

[Correction target item name] 0059

[Correction method] Change

[Correction contents]

On the other hand, as a method of drying and granulating directly from an amino acid fermentation solution using a seed crystal, a solidified material of the amino acid fermentation solution or the above-mentioned amino acid-containing powder is used as a seed crystal, and a fluidized bed granulation is performed. The method of spraying the amino acid-containing liquid into the layer by applying the above method and directly drying and granulating the fermented liquid of the amino acid can be applied. As the seed crystal, the amino acid-containing powder described above can be used. For example, a part of a product obtained by fluidized bed granulation can be pulverized and used as a seed crystal.
In the fluidized-bed granulation, especially by applying the rolling-type fluidized-bed granulation, it is easy to obtain a product having a large bulk density, and it is easy to obtain a granulated product suitable for application to the present invention.

[Procedure amendment 4]

[Document name to be amended] Statement

[Correction target item name] 0072

[Correction method] Change

[Correction contents]

[0072]

[Table 6]

[Procedure amendment 5]

[Document name to be amended] Drawing

[Correction target item name] Fig. 1

[Correction method] Change

[Correction contents]

FIG.

 ──────────────────────────────────────────────────続 き Continuing on the front page (72) Inventor Toshihiko Ishiguri 1-1, Suzukicho, Kawasaki-ku, Kawasaki-shi, Kanagawa Prefecture Ajinomoto Co., Ltd. Production Technology Development Center (72) Inventor Goro Daimon Suzukicho, Kawasaki-ku, Kawasaki-shi, Kanagawa 1-1 Ajinomoto Co., Ltd. Production Technology Development Center F term (reference) 2B150 AC01 AC03 AD02 BB01 BB03 CE16 DA44 DA45 DA48

Claims (4)

[Claims] 1. Cassava potato, peeled, dried and ground to 150 μm or less, is saccharified by suspending it in water to obtain a sugar solution containing a solid residue such as fiber, and fermenting amino acids using this as a fermentation raw material. An additive for feed containing an amino acid as a main component which can be produced and dried and granulated from the obtained amino acid fermentation liquid. 2. An amino acid content of 30 to 80 in terms of dry matter.
The feed additive comprising the amino acid as a main component according to claim 1 in a weight%. 3. The feed additive according to claim 1, wherein the amino acid fermentation liquid is L-lysine, L-threonine or L-tryptophan fermentation liquid. 4. The feed additive according to claim 1, wherein the dry granulation is performed by spray drying granulation.