KR101541007B1 - Process for the preparation of fermentation broth for degrading proteins - Google Patents

Abstract

The present invention relates to a fermentation broth for degrading proteins, obtained by fermenting pineapples, kiwis, and kidney beans. The fermentation broth manufactured according to the present invention acts as a protease, has excellent temperature stability, and has a final product degraded by the fermentation broth containing a large amount of useful amino acids, thereby being useful in recycling a large amount of blood of livestock produced in a slaughter house.

Description

TECHNICAL FIELD [0001] The present invention relates to a fermentation broth for protein degradation,

The present invention relates to a fermentation broth for protein degradation and a method for producing the same, and more particularly, to a fermentation broth for proteolysis that can be useful for decomposition of proteins present in the blood of livestock and a method for producing the same.

The blood of livestock (pigs, cows, etc.) is one of the livestock by-products that are produced in large quantities in slaughterhouses. However, such livestock blood is not utilized as a resource and is mostly disposed of, resulting in waste of valuable protein resources contained in the livestock blood as well as economic loss and environmental destruction due to waste treatment.

In order to reuse the blood of livestock, it is essential to decompose the proteins in the blood of livestock into amino acids, and it is necessary to develop effective proteolytic enzymes.

In this regard, Korean Patent Laid-Open Publication No. 2004-0065201 discloses a method for hydrolyzing a protein by adding a strong acid such as nitric acid or sulfuric acid to an animal's body or body. However, when a strong acid such as nitric acid or sulfuric acid is used, most of the amino acid in the form of a ring is destroyed and only the free amino acid form is present in the decomposition process of the protein. Therefore, the amino acid content of the final product is as low as 2-5% There is a problem that is not continuous and ineffective. Korean Patent Publication No. 1100329 discloses a method for decomposing livestock blood using papain which is a protein hydrolyzing enzyme obtained from papaya fruit milk. However, the enzyme used in the above method requires much time for decomposition, Is not suitable as an enzyme for decomposing proteins in the blood of livestock.

Accordingly, the inventors of the present invention have conducted extensive research to solve the problems of the prior art, and as a result, they have completed the present invention by developing a fermentation broth for protein degradation, which is excellent in protein degradation efficiency and temperature stability.

Korean Patent Publication No. 2004-0065201 Korean Patent Publication No. 1100329

Accordingly, an object of the present invention is to provide a method for producing a fermentation broth for proteolysis, which is excellent in protein degradation efficiency and temperature stability and can be used for treating blood of livestock.

Another object of the present invention is to provide a fermentation broth for proteolysis produced by the above method.

According to an aspect of the present invention, there is provided a method for producing a pineapple comprising the steps of: crushing or cutting pineapple, kiwi and kidney beans; Mixing the crushed or cut pineapple, kiwi and kidney beans with a saccharide; Fermenting the mixture in a fermenter; And filtering the fermented mixture to obtain a fermentation broth.

In order to accomplish the above other objects, the present invention provides a fermentation broth for proteolysis produced by the above production method.

The fermentation broth according to the present invention has excellent protein decomposition efficiency, excellent temperature stability, and is safe and environmentally friendly, obtained from agricultural products. Therefore, the fermentation broth for proteolysis of the present invention can be usefully used in the process for recycling blood of livestock treated with waste.

Hereinafter, the present invention will be described more specifically.

The present invention relates to a method for producing a pineapple comprising the steps of grinding or cutting pineapple, kiwi and kidney beans; Mixing the crushed or cut pineapple, kiwi and kidney beans with a saccharide; Fermenting the mixture in a fermenter; And filtering the fermented mixture to obtain a fermentation broth.

The method for producing the fermentation broth for proteolysis according to the present invention will be described in detail.

<Step 1> Grinding or cutting step

In the grinding or cutting step according to the present invention, pineapples, kiwi and kidney beans are pulverized or cut.

Prior to the above step, the agricultural products may be washed to completely remove pesticides and impurities on the pineapple, kiwi and kidney beans. In the washing, a known detergent may be used to wipe the detergent or an automatic washing machine. Further, in order to completely clean the agricultural products, about 2% of vinegar can be used for the second washing.

The pulverizing or cutting step may be performed by a mechanical pulverizing method, a cutter standard pulverizing method, and a manual pulverizing method. The mechanical pulverization method is a method of pulverizing powder into an apparatus having a rotary knife and powdering it. The above method is effective in shortening the fermentation time and fine filtration after fermentation by pulverizing the fruit or vegetable material as powder. The cutter standard grinding method is a method of setting the standard of each fruit and cutting. Unlike the mechanical grinding method, which does not take into account the characteristics and structure of the fruit, this method can cut to a certain size, preventing excessive crushing of carbohydrates, vitamins, fibers and the like. Therefore, there is an advantage that a large amount of carbohydrates and minerals can be obtained after fermentation. The manual grinding method is a method of directly cutting by hand using a general transient or knife. This method is useful for cutting fruit and seeds containing seeds as compared to other methods, because it can extract seeds of fruits and preserve the chlorophyll of the vegetables.

The pulverized or cut pineapple, kiwi and kidney bean preferably have a size of 0.1 to 30 mm, but are not limited thereto.

&Lt; Second step >

In the mixing step according to the present invention, saccharides are mixed in crushed or cut pineapples, kiwi and kidney beans. Here, the term "saccharide" refers to a carbohydrate that is well soluble in water and has a sweet taste. The saccharides include monosaccharides, disaccharides, and polysaccharides, and include glucose, fructose, maltose, and starch. Specifically, in the present invention, sugar, starch syrup or honey may be used.

The mixture ratio of pineapple: kiwi: kidney bean may be 100 parts by weight of pineapple: 80 to 120 parts by weight of kiwi, 30 to 70 parts by weight of kidney bean, preferably 100 parts by weight of pineapple: 100 parts by weight of kiwi and 50 parts by weight of kidney bean. The saccharide may be added at a ratio of 120 to 140 parts by weight based on 100 parts by weight of the total of pineapple, kiwi and kidney bean. By adding the saccharide at the above ratio, it is possible to prevent the generation of a bad smell caused by the fermentation of acetic acid due to the insufficient sugar content, and the active ingredient contained in the plant can be extracted smoothly.

<Step 3> The fermentation step

In the fermentation step according to the present invention, a mixture of saccharides and pineapple, kiwi and kidney beans mixed in a fermenter is fermented. The fermentation step may be carried out at a temperature of 10 to 40 DEG C for 60 to 365 days after adding the mixture to a stainless steel tank having a magnetic pot or corrosion resistance. By fermenting in the temperature range described above, the activity of the microorganism can be smoothly performed, and the fermentation efficiency can be increased.

In the above process, yeast may be further added to increase the fermentation speed. The yeast may be selected from the group consisting of Saccharomyces cerevisiae , Saccharomyces ellipsoideus , Saccharomyces formosensis , Saccharomyces cerevisiae, and Saccharomyces cerevisiae , which secrete chymotrypsin or trypsin. Karl's Bergen sheath (Saccharomyces carlsbergensis), saccharose in my process mandrel Shurian carcass (Saccharomyces mandshuricus), saccharose in my process Correa Taunus (Saccharomyces coreanus) and may be selected from the group consisting of a combination thereof, to 200, based on the mixture 1L 600 mg. &Lt; / RTI &gt;

<Step 4> Filtration step

In the filtration step according to the present invention, the fermented mixture is filtered to obtain a fermentation broth. The above step may be performed using a centrifugal separator rotating at 3,000 to 10,000 rpm. Finally, the remaining filtrate may be subjected to microfiltration with a filter cloth of 0.5 to 5 μm to remove fine precipitates. As a result, a high purity pineapple, a kiwi and a kidney bean fermentation broth can be obtained.

In addition, the present invention relates to a method for producing a pineapple comprising the steps of: crushing or cutting pineapple, kiwi, kidney bean and strawberry; Mixing the crushed or cut pineapple, kiwi, kidney bean and strawberry with a saccharide; Fermenting the mixture in a fermenter; And filtering the fermented mixture to obtain a fermentation broth. According to the experimental results of the present invention, the fermentation broth in which strawberry is added in addition to pineapple, kiwi and kidney bean has excellent proteolytic ability and amino acid storage ability.

The present invention also provides a fermentation broth for proteolysis, which is produced by the above production method. The proteolytic fermentation broth has a small amount of amino acid breakdown during protein decomposition, is excellent in temperature stability, and is safe and environmentally friendly from agricultural products. Therefore, the fermentation broth for proteolysis of the present invention can be usefully used as a protease in the process of recycling the blood of livestock that is discarded as waste.

Hereinafter, the present invention will be described in more detail with reference to the following examples. The following examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

Example 1: Preparation of fermentation broth using pineapple, kiwi and kidney bean

4 kg of pineapple, 4 kg of kiwi and 2 kg of kidney beans were thoroughly washed in running water and ground to a size of 0.5 mm or less using a cutting machine (PC-14-T, Sung Chang Machinery Co., Ltd.). Then, the ground pineapple, kiwi and kidney beans were placed in a fermentation tank, and 13 kg of sugar was added and mixed. Then, 300 mg / L of Saccharomyces cerevisiae was inoculated into the fermenter and fermented at 35 DEG C for 250 days. The fermented mixture was centrifuged at 5000 rpm for 2 minutes and then filtered with 1 μm filter paper to prepare a high purity fermentation broth.

Comparative Example 1: Preparation of fermentation broth using pineapple and kiwi

A fermentation broth was prepared in the same manner as in Example 1, except that 5 kg of pineapple and 5 kg of kiwi were used instead of 4 kg of pineapple, 4 kg of kiwi and 2 kg of kidney beans.

Comparative Example 2: Preparation of fermentation broth using pineapple and kidney bean

A fermentation broth was prepared in the same manner as in Example 1, except that 4 kg of pineapple, 4 kg of kiwi, and 2 kg of kidney beans were used instead of pineapple (6 kg) and kidney bean (4 kg).

Comparative Example 3: Preparation of fermentation broth using kiwi and kidney beans

A fermentation broth was prepared in the same manner as in Example 1, except that 4 kg of pineapple, 4 kg of kiwi, and 2 kg of kidney beans, 6 kg of kiwi, and 4 kg of kidney beans were used.

Test Example 1: Decomposition of livestock blood using fermentation broth and analysis of components of broken blood

The blood of the pigs produced in the slaughterhouse was collected and completely pulverized for 20 minutes using a blood mill. Then, 30 g of the fermentation broth prepared in Example 1 and Comparative Examples 1 to 3 was added to 1 kg of the pulverized pig blood, and the mixture was allowed to stand at room temperature for 2 hours. The components of the amino acid in the blood were analyzed and are shown in Table 1 below.

                                                     (Unit: wt%) ingredient Example 1 Comparative Example 1 Comparative Example 2 Comparative Example 3 Aspartic acid 1.95 1.45 1.90 1.81 Threonine 0.96 0.52 0.61 0.51 Serine 0.82 0.52 0.79 0.77 Glutamic acid 1.57 0.64 1.31 1.19 Proline 0.94 0.57 0.93 0.82 Glycine 0.82 0.50 0.67 0.74 Alanine 1.12 0.67 1.02 0.91 Balin 1.28 0.87 1.17 1.14 Isoleucine 0.19 0.09 0.09 0.17 Leucine 2.17 1.46 1.91 1.72 Tyrosine 0.39 0.25 0.29 0.24 Phenylalanine 0.97 0.65 0.83 1.01 Histidine 0.93 0.56 0.88 0.83 Lysine 1.07 0.75 0.95 0.92 Arginine 0.60 0.27 0.41 0.44 Cystine 0.25 0.13 0.23 0.18 Methionine 0.14 0.04 0.12 0.10 Tryptophan 0.17 0.12 0.14 0.09 Sum 16.34 10.06 13.25 13.59

As shown in Table 1, the blood decomposed using the fermentation broth of Example 1 contained a large amount of 18 amino acids compared with the blood decomposed using the fermentation broths of Comparative Examples 1 to 3. The fermentation broths of Example 1, Comparative Example 2 and Comparative Example 3 prepared by adding kidney bean showed better proteolytic ability and amino acid storage ability than the fermentation broth of Comparative Example 1 in which kidney beans were not added .

These results show that the addition of kidney beans to the fermentation broth improves the content of amino acids in the degraded blood. In particular, the use of the fermentation broth of pineapple, kiwi and kidney bean as degrading enzymes results in an increase in the amino acid content The most effective for heightening the

Test Example  2: Analysis of components of broken blood

Using the fermentation broth of Example 1 and a commercially available protease, the blood of the livestock was decomposed in the same manner as in Test Example 1, and then the amino acid components in the degraded blood were compared. The results are shown in Table 2 below.

                                                          (Unit: wt%) ingredient Example 1 Bromelain
(Bromelain 1000, JARROW) Actinidine
(Mega actinidin, TWINLAB) Papain
(Plant Enzyme of Papain, NOW) Quercetin
(Quercetin with Bromelain, NOW) Aspartic acid 1.95 1.45 1.12 1.11 1.90 Threonine 0.96 0.92 0.56 0.71 0.92 Serine 0.82 0.54 0.80 0.67 0.42 Glutamic acid 1.57 0.94 1.00 1.49 1.19 Proline 0.94 0.87 0.84 0.82 0.57 Glycine 0.82 0.50 0.63 0.24 0.49 Alanine 1.12 0.67 0.77 0.90 0.81 Balin 1.28 0.87 1.01 1.09 0.97 Isoleucine 0.19 0.29 0.09 0.10 0.19 Leucine 2.17 1.46 1.41 1.27 1.49 Tyrosine 0.39 0.45 0.33 0.33 0.45 Phenylalanine 0.97 0.61 0.74 1.01 0.85 Histidine 0.93 0.58 0.88 0.81 0.76 Lysine 1.07 0.75 0.94 0.90 0.95 Arginine 0.60 0.27 0.40 0.44 0.47 Cystine 0.25 0.14 0.27 0.17 0.12 Methionine 0.14 0.05 0.13 0.12 0.26 Tryptophan 0.17 0.17 0.17 0.09 0.23 Sum 16.34 11.53 12.09 12.27 13.04

As shown in Table 2, it can be seen that a large amount of amino acid is contained in the blood digested with the fermentation broth of Example 1 as compared with the commercially available proteases.

The above results show that the fermentation broth according to the present invention shows better proteolytic efficiency than the conventional proteolytic enzyme, and thus the fermentation broth of the present invention shows remarkably excellent effect as a proteinase of livestock blood to be discarded.

Test Example  3: Evaluation of temperature stability

The fermentation broth of the fermentation broth prepared in Example 1 and Comparative Examples 1 to 3 was examined for the enzyme activity at 5 ° C, 15 ° C, 25 ° C, 35 ° C, 45 ° C, 55 ° C and 65 ° C, After reacting with casein for 10 minutes each, enzymatic activity was measured by azo casein analysis. The higher the enzyme activity rate (%), the higher the activity at the temperature, and the optimal activity range was determined to be over 70% of the objective criterion. The results are shown in Table 3 below.

Temperature 5 ℃ 15 ℃ 25 ℃ 35 ℃ 45 ° C 55 ° C 65 ℃ Example 1 14 63 84 98 89 71 24 Comparative Example 1 14 22 37 91 41 27 11 Comparative Example 2 14 35 61 98 72 41 16 Comparative Example 3 14 39 57 96 68 52 13

As shown in Table 3, the fermentation broth of Example 1 showed optimum activity at 25 to 55 ° C, Comparative Example 2 at 35 to 45 ° C, and Comparative Examples 1 and 3 at 35 ° C only. This shows that the fermentation broth of the present invention has a wide range of temperatures that can be used compared to the fermentation broths of Comparative Examples 1 to 3, and thus the fermentation broth of the present invention can be applied in various conditions when treating livestock blood .

Example 2: Preparation of fermentation broth using pineapple, kiwi, kidney bean and strawberry

A fermentation broth was prepared in the same manner as in Example 1 except that 3 kg of pineapple, 3 kg of kiwi, 2 kg of strawberry and 2 kg of kidney bean were used instead of 4 kg of pineapple, 4 kg of kiwi and 2 kg of kidney beans.

Test Example 4: Analysis of components of degraded blood

The blood components of the livestock degraded using the fermentation broths prepared in Examples 1 and 2 were analyzed and shown in Table 4 below.

                                                   (Unit: wt%) ingredient Example 1 Example 2 Aspartic acid 1.95 1.94 Threonine 0.96 0.92 Serine 0.82 0.89 Glutamic acid 1.57 1.71 Proline 0.94 0.94 Glycine 0.82 0.97 Alanine 1.12 1.27 Balin 1.28 1.42 Isoleucine 0.19 0.21 Leucine 2.17 2.26 Tyrosine 0.39 0.55 Phenylalanine 0.97 0.98 Histidine 0.93 1.04 Lysine 1.07 1.09 Arginine 0.60 0.63 Cystine 0.25 0.27 Methionine 0.14 0.14 Tryptophan 0.17 0.19 Sum 16.34 17.42

As shown in Table 4, it can be seen that the blood decomposed using the fermentation broth of Example 2 contained a large amount of amino acids compared with the blood decomposed using Example 1. [

Claims (7)

1) crushing or cutting pineapple, kiwi and kidney beans;
2) mixing the pulverized or cut pineapple, kiwi and kidney beans with a saccharide;
3) fermenting the mixture in a fermentation tank; And
4) filtering the fermented mixture to obtain a fermentation broth.
The method according to claim 1,
Wherein the mixing ratio of the pineapple: kiwi: kidney beans is 100 parts by weight of pineapple: 80 to 120 parts by weight of kiwi: 30 to 70 parts by weight of kidney beans.
The method according to claim 1,
Wherein the saccharide of step 2) is added in a proportion of 120 to 140 parts by weight based on 100 parts by weight of the total of pineapple, kiwi and kidney bean.
The method according to claim 1,
Three saccharide in my process to the step 3) Levy jiae (Saccharomyces cerevisiae), my process as Saccharomyces ellipsis Soy Ranges (Saccharomyces ellipsoideus), saccharose in my process formate Mohsen sheath (Saccharomyces formosensis), a saccharide My process car's Bergen sheath (Saccharomyces carlsbergensis), a saccharide My process mandrel Shurian carcass (Saccharomyces mandshuricus), a saccharide My process Correa Taunus (Saccharomyces coreanus ), and a combination thereof. &lt; Desc / Clms Page number 20 &gt;
The method according to claim 1,
Wherein the fermentation of step 3) is carried out at a temperature of 10 to 40 DEG C for 60 to 365 days.
1) grinding or cutting pineapple, kiwi, kidney bean and strawberry;
2) mixing the pulverized or cut pineapple, kiwi, kidney bean and strawberry with a saccharide;
3) fermenting the mixture in a fermentation tank; And
4) filtering the fermented mixture to obtain a fermentation broth.
A fermentation broth for proteolysis, which is produced by the method of any one of claims 1 to 6.