KR20100115043A - Fibrinolytic composition having culture of bacillus subtilis natto and mushroom fungi - Google Patents

Abstract

PURPOSE: A composition containing Bacillus subtilis natto culture and mushroom bacteria culture for thrombolysis is provided to enhance thrombolytic ability. CONSTITUTION: A composition for thrombolysis contains Bacillus subtilis natto culture and mushroom bacteria culture in a weight ratio of 100:1-2. The mushroom culture is mycelium culture of Agaricus blazei, Pleurotus ostreatus, Ganoderma lucidum, Lentinus edodes or Coriolus versicolor.

Description

Fibrinolytic Composition Having Culture Of Bacillus Subtilis Natto And Mushroom Fungi}

The present invention relates to a composition having fibrinolytic activity, in particular to a composition for dissolving a blood clot comprising a Bacillus subtilis natto culture and a mushroom culture, in particular by a mushroom culture The present invention relates to a composition capable of significantly increasing the thrombolytic ability of natto bacteria.

The production of blood clots (small blood clots in the blood vessels) in the blood vessels leads to poor blood flow and symptoms of cardiovascular diseases such as increased blood pressure and stroke, which are very common for modern people who eat a lot of fat. It is a serious problem. Thrombosis and artherosclerosis are caused by the interaction of excessive platelet aggregation, blood coagulation and blood lipids, and hypertension and smoking are known risk factors for atherosclerosis.

In vivo, blood has always been balanced in coagulation and lysis, and clots generated during normal circulation are readily degraded (Arbige and Pitcher, 1989). A thrombus is produced when a blood component, fibrinogen, is converted to fibrin by thrombin to form an insoluble polymer when the body is injured. When blood clots are formed in veins, blood circulation disorders occur, causing swelling and inflammation, but when they occur in arteries, they cause ischemia or infarction, leading to cardiovascular diseases such as arteriosclerosis, myocardial infarction, stroke and pulmonary embolism [Grande et al. 1990; Ruggeri and Zimmerman 1985; Sherman and Hart 1986; Shin et al. 2007].

In order to prevent thrombus formation, anti-coagulants are required, and in the case of the formed thrombi, fibrin clot disintegrator is required to dissolve the thrombus and treat the thrombus [Sherman and Hart, 1986]. Thrombolytics are classified into thrombolytic enzymes and plasminogen activator (PA). PA acts on plasminogen in the blood to produce plasmin, and plasmin includes urokinase (UK) [Toki et al., 1985], streptokinase [Fletcher and Johnson, 1957; Lijnen et al., 1992], staphylokinase [Lijnen et al., 1992], nattokinase (NK) [Sumi et al., 1990], tissue type plasminogen activator (tPA) [Astrup and Sterndorff, 1956], and the like.

Nattokinase is a type of microbial serine protease that is absorbed into the body during oral administration and acts as a plasminogen activator to convert plasminogen into plasmin, which has fibrinolytic activity by dissolving fibrin in the coagulation blood [Kim, 2002; Sumi 1990; Yang 2000; Yong et al. 2005]. Natokinase not only directly degrades closs-linked fibrin in vitro , but also promotes tPA production, regulates total thrombolytic activity by the relative ratio of tPA, and is the primary inhibitor of plasminogen activator inhibitor, fibrin clot lysis. Enhances thrombolysis by inactivation and degradation of -1 (PAI-1) [Fujita et al. 1995; Urano et al. 2001].

Anticoagulants, antiplatelets, thrombolytics such as heparin, qumarin, asprin UK are used for the prevention and treatment of thrombotic diseases, but they are not only expensive but also due to problems such as hemorrhagic side effects, gastrointestinal disorders, irritability and parenteral administration. Are limited [Gwak and Chun, 2000; Lee et al., 2002; Sohn et al., 2005; Sumi et al., 1989; Weitz and Crowther, 2002]. To compensate for these drawbacks, Bacillus sp . Research on the characteristics of the derived thrombolytic enzymes has been actively conducted recently.

In general, when the soybeans are fermented naturally with rice straw (30-40C), fermented bacteria that produce a transparent viscous substance are inhabited, which is Bacillus subtilis natto. Bacillus subtilis natto is listed in the ATCC is registered seven species, there are a variety of strains reported separately from natural fermentation. In Korea, it has been used as a food for long period of time as a Cheonggukjang and as a NATO in Japan.In recent years, it has been used as a food or food additive not only for Asians but also for the purpose of health promotion. have.

The thrombolytic action of the various physiologically active functions of these natto bacteria has already been revealed from various studies.

However, when preparing a functional food using the above-described natto bacteria, the thrombolytic ability by the natto bacteria was insufficient, and thus, in the preparation of the functional food using the natto bacteria, the natto bacteria were treated. There is also an urgent need for various methods of processing and other methods that can further increase the thrombolytic ability by the natto bacteria.

The present invention for solving the above problems is to provide a new composition that can increase the thrombolytic ability of natto bacteria.

In addition, the present invention is to include the mushroom bacteria together with the NATO bacteria, it is an object to significantly increase the thrombolytic ability of the NATO bacteria by the mushroom bacteria.

In addition, the present invention is to provide a powder-like composition that can be used simply and easily in the process of manufacturing a functional food by a specific method of processing and processing the natto bacteria in order to use the natto bacteria in the production of functional food. do.

The present invention for achieving the above object is a composition for dissolving a thrombus containing a natto bacteria culture and mushroom culture.

Here, the natto bacteria culture may be a culture in which Bacillus subtilis natto bacteria are cultured, and the mushroom culture may be a culture in which mushroom mycelium is cultured. Particularly, among the various mushrooms, cultures of mycelium using Agaricus blazei , Pleurotus ostreatus , Ganoderma lucidum , Shiitake ( Lentinus edodes ), or shiitake mushroom ( Coriolus versicolor ) are most preferred. .

In order to further enhance the fibrinolytic activity of natto or nattokinase, the present inventors have mixed mushrooms with natto bacteria, and the mixed mixture has remarkably superior thrombolytic activity than that containing only natto bacteria. After confirming that, this invention was completed. In particular, natto bacteria and mushrooms were prepared by culturing these cultures, respectively, and then mixed, thereby easily and easily preparing a mixed composition and simultaneously synergistic effects between the two components.

In addition, the natto bacteria culture and the mushroom culture is preferably included in a weight ratio of 100: 1 to 2, which is insufficient to enhance the thrombolytic activity of the natto bacteria culture when outside the above range, or further included. Even if the thrombolytic activity does not increase significantly.

Furthermore, the above-described mixed composition according to the present invention may be prepared in powder form by preparing each of the natto bacteria cultures and mushroom mycelium cultures in a concentrated state, and then spray-drying a mixture thereof. By concentrating each of the cultures and mushroom cultures and preparing powdered powdered powder, it can be conveniently used in the production of functional foods, and the effect can be quickly exhibited even when the functional foods are ingested.

Specific details of other embodiments are included in the detailed description and the drawings.

The composition for thrombolytic dissolution according to the present invention includes mushrooms together with natto bacteria, thereby increasing the blood clot dissolving ability of natto bacteria by the mushrooms.

In particular, by using a culture in which the natto and mushroom bacteria were cultured, and in addition, by powdering the concentrated solution concentrated in the culture, a powder-like composition that can be used simply and easily in the process of producing a functional food There is an effect that can be provided.

In addition, the present invention, when using a mycelium of Agaricus blazei , Pleurotus ostreatus , Ganoderma lucidum , Lentinus edodes or Shiitake mushroom ( Coriolus versicolor ), among many mushrooms, There is an effect that can significantly increase the thrombolytic ability of natto bacteria more significantly.

Hereinafter, one preferred embodiment of the present invention will be described in detail with reference to the accompanying drawings. The invention may be better understood by the following examples, which are intended for purposes of illustration of the invention and are not intended to limit the scope of protection defined by the appended claims.

Example 1 Preparation of Natobacterial Culture Concentrate Powders

(1) Preparation of nattokinase concentrate (NKC)

5 L of Bacillus subtilis natto obtained from Korea Microorganism Conservation Center (KCCM) using sterilized liquid medium (121C, 1 hour) containing skim soybean powder (5%) and maltose (1%) Incubator for 48 hours (30 ℃) in a dragon incubator (Kobiotech, Co., Inchen, Korea), filtered and concentrated to more than 5,000U / ml Fibrinolytic activity. The specific process is as shown in Table 1 below.

Table 1: Preparation of Nato bacteria culture concentrate

Manufacture process fair Remarks Feeding medium into the incubator (5KL) Medium component: Defatted soy flour 5%, maltose 1%   ↓ Sterilization Sterilization temperature and time: 121C, 1 hour   ↓ Medium cooling Cooling temperature: 25 ℃  ↓ Seed culture inoculation Strains: Bacillus subtilis natto KCCM12027 ↓ culture Incubation temperature: 30 ℃, Incubation time: 48 hours ↓ percolation Filter pore size: 0.8um ↓ concentration Fibrinolytic activity: more than 5,000U / ml

(2) Powdering of Natobacterial Culture Concentrate

 By adjusting the indigestible maltodextrin to the concentrated Nato bacteria culture concentrate was adjusted to a solid content of 35%, and then spray-dryed (first pore, J-004) to prepare a powder.

Example 2 Preparation of Powder of Mushroom Mycelia Culture Extract Concentrate (Brix 30)

1) Medium Preparation

Degreased soy flour (20 g), 20 g per sulfur white, 0.5 g MgSO _4, 0.5 g KH ₂ PO ₄ was mixed with 1 L of water, and sterilized at 121 ° C. for 30 minutes to use as a liquid medium.

2) Culture

After inoculating each of the mushroom bacteria of the following [Table 2] in the liquid medium (cultivation conditions: 25C, 1v / v / m airation).

Table 2: Types of Mushroom Strains

Abbreviation Common name Scientific name AB   Agaricus Mushroom Agaricus blazei PO  Oyster mushroom Pleurotus ostreatus GL  Ganoderma lucidum mushroom  Ganoderma lucidum LE Shiitake mushrooms Lentinus edodes PJ  Cordyceps sinensis Paecilomyces japonicus PL   Situation Mushroom Phellinus linteus GF  Leaf mushroom Grifola frondosa CV   Fingering mushroom Coriolus versicolor

3) Preparation of mushroom mycelium culture extract concentrate (Brix 30)

The culture was incubated at 121C for 1 hour under high pressure, and then filtered (membrane filter pore size 0.8um) was concentrated to Brix 30.

4) Powdered Mushroom Mycelia Culture Extract Concentrate (Brix 30)

The refractory maltodextrin was mixed with the concentrated mushroom mycelium culture extract concentrate (Brix 30), adjusted to a solid content of 35%, and then spray dried (first pore, J-004) to prepare a powder.

Example 3 Preparation of Mixed Powder of NATO Culture and Mushroom Mycelia Culture

1 kg of the mushroom mycelium culture concentrate (Brix 30) prepared in [Example 2] was mixed with the natto bacteria culture (100 kg) prepared in Example 1, followed by mixing an indigestible maltodextrin in these mixtures. The solid content was adjusted to 35%, followed by spray drying (first pore, J-004) to prepare a powder.

Experimental Example 1 Measurement of Thrombolytic Activity

1) Experiment

In the present experiment, the sample prepared in [Example 1], that is, the powdered sample of NATO culture concentrate (NKCP), and the sample prepared in [Example 2], that is, mushroom mycelium liquid culture extract concentrate (Brix 30) ), A mixture of a powdered sample (AB, PO, GL, LE, PJ, PL, GF, CV) and the sample prepared in Example 3, that is, a NATO culture concentrate and a mushroom mycelium culture extract concentrate. (NKF; NKCP-AB, NKCP-PO, NKCP-GL, NKCP-LE, NKCP-PJ, NKCP-PL, NKCP-GF, NKCP-CV) was tested for the effect of enhancing the thrombolytic activity of the powdered samples.

2) Method of measuring thrombolytic activity

The thrombolytic activity of each sample prepared from the powder was measured by the following method by modifying the method of Astrup and Mullertz (1952).

Preparation of Test Solution

* Sodium borate buffer solution preparation: 50mM sodium borate solution [sodium tetraborate decahydride 19.07g / L] and 0.2M Boric acid [Boric acid 12.366g / L] were prepared, respectively, 50mM Sodium borate solution and 0.2M Boric acid 1: Mix in 4 ratios.

* 0.2M Trichloroacetic acid (TCA) solution preparation: 32.678g Trichloroacetic acid (TCA) is applied to 1L distilled water.

* Preparation of Thrombin solution: Dissolve 51.95 ml of Sodium borate buffer solution in Thrombin (1,039 NIH units; Sigma-Aldrich, T1063-1KU), dissolve (20 units / ml), and dispense 0.4 ml each into E-tube.

* 0.5% Fibrinogen solution preparation: Add 1.6ml of Sodium borate buffer solution to Fibrinogen 12.5mg (Fluka no. 46313; Purity 64%) and mix thoroughly.

* Preparation of Fibrin Mixture: Add 25μl of Thrombin solution, 100μl of 0.5% Fibrinogen solution, and 150μl Sodium borate buffer solution to E-tube (2ml volume), close the lid and leave at 37 ℃ for 10 minutes. In this case, four E-tubes are prepared, including three repetitions and a blank to analyze one sample.

Thrombolytic activity measurement

① Add 200 ㎕ of sodium borate buffer solution to the fibrin admixture and leave at room temperature for 5-15 minutes after stirring.

② Add 25 g of the diluted solution of 100 g of sodium borate buffer solution to the fibrin diluent and stir.

③ Leave at 37 ° C. for 30 minutes and then stir for 5 seconds.

④ Leave at 37 ° C. for 30 minutes.

⑤ Add 0.2M TCA 0.5ml to each E-tube and mix lightly to stop the reaction.

⑥ Leave at room temperature for 20 minutes.

⑦ Centrifuge (15,000 rpm, 10 minutes).

⑧ Measure the absorbance of the supernatant (275nm).

⑨ Calculate the enzyme activity by the following formula.

Thrombolytic activity (U / mL) =

  At: Absorbance r value of the sample (275nm)

  Ab: Absorption value of blank test sample (blank) prepared without putting sample

  0.01: activity of enzyme with absorbance increased by 0.01 for 1 minute

  60: enzyme reaction time (min)

  1 / 0.1 to 1: total reaction solution, 0.1: fibrinogen content in reaction solution

  D: dilution factor

3) Experiment result

The results measured as described above are as shown in the following [Table 3] to [Table 10]. That is, the thrombolytic effect of natto bacteria cultures was increased by mushroom variety.

In particular, agaricus mushroom, oyster mushroom, ganoderma lucidum mushroom, shiitake mushroom, and fingerling mushroom among the strains used in this experiment showed a significant enhancement effect compared to the NKCP powdered sample (NKCP).

Table 3: Effect of Agaricus Mushroom Mycelia Extract on Enhancement of Thrombolytic Effect of Nato Bacteria Culture

Category ¹⁾ Thrombolytic activity ²⁾
(Fibrinolytic activity) NKCP 17,022 ± 385 ^{b 3)} AB 4,722 ± 51 ^c NKCP-AB 23,132 ± 415 ^a

¹⁾ NKCP: Mix the natto bacteria culture concentrate prepared in Example 1 with an indigestible maltodextrin to adjust the solid content to 35%, and then spray dry the sample, AB: Agaricus mushroom mycelium cultured extract concentrate (Brix 30 ), Mixed with indigestible maltodextrin to adjust the solid content to 35%, and then spray-dried sample, NKCP-AB: 1kg of Agaricus mushroom mycelium culture culture extract extract (Brix 30) in Nato bacteria culture concentrate (100kg) Mix with maltodextrin to adjust the solids to 35% and then spray dry the sample

²⁾ Average of 3 replicate analysis of 5 lots samples

³⁾ Statistical processing: 0.05% significant difference by Duncan's multiple test (other lowercase letters are significant)

Table 4: Effect of Pleurotus eryngii Extract on Enhancement of Thrombolytic Effect of Nato Bacteria Culture]

Category ¹⁾ Thrombolytic activity ²⁾
(Fibrinolytic activity) NKCP 17,022 ± 385 ^{b 3)} PO 4,080 ± 43 ^c NKCP-PO 22,233 ± 348 ^a

¹⁾ NKCP: A mixture of the NATO culture concentrate prepared in [Example 1] with an indigestible maltodextrin was adjusted to a solid content of 35%, followed by spray dry sample, PO: Oyster mushroom mycelium cultured extract concentrate (Brix 30 ), Mixed with indigestible maltodextrin to adjust the solid content to 35%, and then spray-dried sample, NKCP-AB: 1 kg of oyster mushroom mycelium culture extract extract concentrate (Brix 30) was added to Nato bacteria culture concentrate (100 kg). Mix with maltodextrin to adjust the solids to 35% and then spray dry the sample

²⁾ Average of 3 replicate analysis of 5 lots samples

³⁾ Statistical processing: 0.05% significant difference by Duncan's multiple test (other lowercase letters are significant)

[Table 5: Addition of Ganoderma lucidum mycelium extract to increase the thrombolytic effect of natto bacteria culture]

Category ¹⁾ Thrombolytic activity ²⁾
(Fibrinolytic activity) NKCP 17,022 ± 385 ^{b 3)} GL 3,162 ± 74 ^c NKCP-GL 22,893 ± 521 ^a

¹⁾ NKCP: A mixture of the NATO culture concentrate prepared in [Example 1] with an indigestible maltodextrin was adjusted to a solid content of 35%, followed by spray dry sample, GL: Ganoderma lucidum mycelium cultured extract concentrate (Brix 30 ), Mixed with indigestible maltodextrin to adjust the solid content to 35%, and then spray-dried sample, NKCP-GL: 1kg of Ganoderma lucidum mycelium culture culture extract concentrate (Brix 30) 1kg into natto bacteria culture concentrate (100kg) Mix with maltodextrin, adjust the solids to 35%, and then spray dry the sample,

²⁾ Average of 3 replicate analysis of 5 lots samples

³⁾ Statistical processing: 0.05% significant difference by Duncan's multiple test (other lowercase letters are significant)

Table 6: Effect of Shiitake Mushroom Mycelia Extract on Enhancement of Thrombolytic Effect of Nato Bacteria Culture]

Category ¹⁾ Thrombolytic activity ²⁾
(Fibrinolytic activity) NKCP 17,022 ± 385 ^{b 3)} LE 3,583 ± 92 ^c NKCP-LE 21,552 ± 327 ^a

¹⁾ NKCP: A mixture of the NATO culture concentrate prepared in Example 1 was mixed with an indigestible maltodextrin to adjust the solid content to 35%, followed by spray dry sample, LE: Shiitake mycelium cultured extract concentrate (Brix 30 ), Mixed with indigestible maltodextrin to adjust the solid content to 35%, and then spray-dried sample, NKCP-LE: 1kg of shiitake mushroom mycelium culture culture extract concentrate (Brix 30) in indigestion of natto bacteria culture concentrate (100kg) Mix with maltodextrin to adjust the solids to 35% and then spray dry the sample

²⁾ Average of 3 replicate analysis of 5 lots samples

³⁾ Statistical processing: 0.05% significant difference by Duncan's multiple test (other lowercase letters are significant)

Table 7: Addition of Cordyceps Sinensis Mycelia Extracts to Enhance the Thrombolytic Effect of Nato Bacteria Culture]

Category ¹⁾ Thrombolytic activity ²⁾
(Fibrinolytic activity) NKCP 17,022 ± 385 ^{a 3)} PJ 1,984 ± 82 ^b NKCP-PJ 17,433 ± 593 ^a

¹⁾ NKCP: Mix the natto bacteria culture concentrate prepared in Example 1 with an indigestible maltodextrin to adjust the solid content to 35% and then spray dry the sample, PJ: Cordyceps fungus mycelium cultured extract concentrate (Brix 30 ), Mixed with indigestible maltodextrin to adjust the solid content to 35%, and then spray-dried sample, NKCP-PJ: Nato bacteria culture concentrate (100 kg), 1 kg of Cordyceps fungus mycelium culture extract concentrate (Brix 30) Mix with maltodextrin to adjust the solids to 35% and then spray dry the sample

²⁾ Average of 3 replicate analysis of 5 lots samples

³⁾ Statistical processing: 0.05% significant difference by Duncan's multiple test (other lowercase letters are significant)

Table 8: Effect of Situation Mushroom Mycelia Extract on Enhancement of Thrombolytic Effect of Nato Bacteria Culture]

division Thrombolytic activity ²⁾
(Fibrinolytic activity) NKCP 17,022 ± 385 ^{a 3)} PL 991 ± 54 ^b NKCP-PL 17,367 ± 645 ^a

¹⁾ NKCP: A mixture of NATO culture concentrate prepared in [Example 1] with an indigestible maltodextrin was adjusted to a solid content of 35%, followed by spray dry sample, PL: Situary mushroom mycelium cultured extract concentrate (Brix 30 ), Mixed with indigestible maltodextrin to adjust the solid content to 35%, and then spray-dried sample, NKCP-PL: Nato bacteria culture concentrate (100 kg), 1 kg of the situation mushroom mycelia culture extract concentrate (Brix 30) Mix with maltodextrin to adjust the solids to 35% and then spray dry the sample

²⁾ Average of 3 replicate analysis of 5 lots samples

³⁾ Statistical processing: 0.05% significant difference by Duncan's multiple test (other lowercase letters are significant)

[Table 9: Addition of leaf fungus mycelium extract to increase the thrombolytic effect of natto bacteria culture]

Category ¹⁾ Thrombolytic activity ²⁾
(Fibrinolytic activity) NKCP 17,022 ± 385 ^{a 3)} GF 1,725 ± 62 ^b NKCP-GF 17,120 ± 425 ^a

¹⁾ NKCP: The NATO culture concentrate prepared in Example 1 was mixed with an indigestible maltodextrin to adjust the solid content to 35%, followed by spray dry sample, GF: leaf fungus mycelium cultured extract concentrate (Brix 30 ), Mixed with indigestible maltodextrin to adjust the solid content to 35%, and then spray-dried sample, NKCP-GF: Nato bacteria culture concentrate (100 kg), 1 kg of leaf fungus mycelium culture extract concentrate (Brix 30) Mix with maltodextrin to adjust the solids to 35% and then spray dry the sample

²⁾ Average of 3 replicate analysis of 5 lots samples

³⁾ Statistical processing: 0.05% significant difference by Duncan's multiple test (other lowercase letters are significant)

[Table 10: Addition of fingertip mushroom mycelium extract to enhance the thrombolytic effect of natto bacteria culture]

Category ¹⁾ Thrombolytic activity ²⁾
(Fibrinolytic activity) NKCP 17,022 ± 385 ^{b 3)} CV 3,678 ± 75 ^c NKCP-CV 21,334 ± 378 ^a

¹⁾ NKCP: A mixture of the NATO culture concentrate prepared in Example 1 was mixed with an indigestible maltodextrin and adjusted to a solid content of 35%, followed by spray dry sample, CV: fingerling mushroom mycelium cultured extract concentrate (Brix 30). ), Mixed with indigestible maltodextrin to adjust the solid content to 35%, and then spray-dried sample, NKCP-CV: Nato bacteria culture concentrate (100 kg), 1 kg of fingertip mushroom mycelium culture extract concentrate (Brix 30) Mix with maltodextrin to adjust the solids to 35% and then spray dry the sample

²⁾ Average of 3 replicate analysis of 5 lots samples

³⁾ Statistical processing: 0.05% significant difference by Duncan's multiple test (other lowercase letters are significant)

Experimental Example 2 Measurement of Thrombolytic Activity According to the Concentration of Mushroom Culture

1) Experiment

Experimental Example 1, in the mixed composition comprising the Ganoderma lucidum culture and Agaricus mushroom culture, respectively, confirmed to have excellent thrombolytic activity, the concentration of the Ganoderma lucidum culture and Agaricus mushroom culture, that is, according to the mixing ratio To measure the thrombolytic activity.

To this end, the concentration of the Ganoderma lucidum culture and Agaricus mushroom culture (Brix 30) of the Ganoderma lucidum culture prepared in [Example 2] was 0.5, 1, and 2 in the NATO culture (100 kg) prepared in [Example 1]. , 3, 4, and 5kg each was mixed, and then mixed with an indigestible maltodextrin to these mixtures to adjust the solid content to 35% and then spray dry (first pore, J-004) to prepare a powder.

The thrombolytic enzyme activity was measured in the same manner as in [Experimental Example 1], and the results are shown in the following [Table 11] and [Table 12].

2) Experiment result

As a result, as can be seen in Table 11 and Table 12, when 0.5 kg of mushroom culture was mixed (that is, a ratio of 100: 0.5), the increase in thrombolytic activity was insufficient, and 3 kg (that is, , The ratio of thrombolytic activity does not increase significantly even if the ratio is higher than 100: 3.

[Table 11: Effects of Enhancement of Thrombolytic Activity According to Addition of Agaricus Mushroom Mycelia Extract by Concentration]

Category ¹⁾ Thrombolytic activity ²⁾
(Fibrinolytic activity) NKCP 17,022 ± 385 ^{b 3)} AB 4,722 ± 51 ^c NKCP-AB 0.5 18,332 ± 195 ^b NKCP-AB 1 23,132 ± 415 ^a NKCP-AB 2 24,352 ± 432 ^a NKCP-AB 3 24,300 ± 359 ^a NKCP-AB 4 24,122 ± 324 ^a NKCP-AB 5 24,532 ± 532 ^a

¹⁾ NKCP: Mix the natto bacteria culture concentrate prepared in Example 1 with an indigestible maltodextrin to adjust the solid content to 35%, and then spray dry the sample, AB: Agaricus mushroom mycelium cultured extract concentrate (Brix 30 ), Mixed with indigestible maltodextrin to adjust the solid content to 35%, and then spray-dried sample, NKCP-AB 0.5: 0.5 kg of Agaricus mycelium culture culture extract concentrate (Brix 30) to NATO culture concentrate (100 kg) After mixing with indigestible maltodextrin and adjusting the solid content to 35%, spray dried sample, NKCP-AB 1: Agaricus mycelium mycelium culture culture extract concentrate (Brix 30) 1kg into natto bacteria culture concentrate (100kg) After mixing with dextrin and adjusting the solid content to 35%, spray dried sample, NKCP-AB 2: Nato bacteria culture concentrate (100kg), 2kg Agaricus mushroom mycelium culture extract concentrate (Brix 30) After mixing with maltodextrin to adjust the solid content to 35%, spray dried sample, NKCP-AB 3: Agaricus mushroom mycelium culture culture extract concentrate (Brix 30) 3kg to the indigestible maltodextrin After mixing to adjust the solid content to 35%, spray-dried sample, NKCP-AB 4: Nato bacteria culture concentrate (100 kg), 4 kg Agaricus mushroom mycelium culture extract extract (Brix 30) mixed with indigestible maltodextrin solids After adjusting to 35% and spray-drying the sample, NKCP-AB 5: Natobacterial culture concentrate (100kg), 5kg of Agaricus mushroom mycelium culture extraction extract (Brix 30) was mixed with indigestible maltodextrin and the solid content was 35%. Sample to be spray dried

²⁾ Average of 3 replicate analysis of 5 lots samples

³⁾ Statistical processing: 0.05% significant difference by Duncan's multiple test (other lowercase letters are significant)

[Table 12: Effects of Ganoderma lucidum Mycelium Extract on the Enhancement of Thrombolytic Effects of Nato Bacteria Culture]

Category ¹⁾ Thrombolytic activity ²⁾
(Fibrinolytic activity) NKCP 17,022 ± 385 ^{b 3)} GL 3,162 ± 74 ^c NKCP-GL 0.5 18,260 ± 152 ^b NKCP-GL 1 22,893 ± 521 ^a NKCP-GL 2 23,175 ± 253 ^a NKCP-GL 3 24,365 ± 152 ^a NKCP-GL 4 24,198 ± 160 ^a NKCP-GL 5 24,217 ± 231 ^a

¹⁾ NKCP: A mixture of the NATO culture concentrate prepared in Example 1 was mixed with an indigestible maltodextrin to adjust the solid content to 35%, followed by spray dry sample, GL: Ganoderma lucidum mycelium cultured extract concentrate (Brix 30 ) Mixed with indigestible maltodextrin to adjust the solid content to 35%, and then spray-dried sample, NKCP-GL 0.5: Nato bacteria culture concentrate (100 kg) to 0.5 kg of Ganoderma lucidum mycelium culture extract concentrate (Brix 30) After mixing with indigestible maltodextrin and adjusting the solid content to 35%, spray dried sample, NKCP-GL 1: Nato bacteria culture concentrate (100 kg), 1 kg of Ganoderma lucidum mycelium culture extract concentrate (Brix 30) was added to indigestible maltose After mixing with dextrin to adjust the solids to 35% and spray-drying the sample, NKCP-GL 2: Nato bacteria culture concentrate (100 kg) and 2 kg of Ganoderma lucidum mycelium culture extract extract concentrate (Brix 30) with indigestible maltodextrin So After adjusting the solids to 35% and spray-drying the sample, NKCP-GL 3: 3 kg of Ganoderma lucidum mycelium culture culture extract concentrate (Brix 30) was mixed with the indigestible maltodextrin in a concentrated solution of NATO bacteria (100 kg). After adjusting to%, spray dried sample, NKCP-GL 4: Nato bacteria culture concentrate (100 kg), mixed with 4 kg of Ganoderma lucidum mycelium culture extract extraction concentrate (Brix 30) with indigestible maltodextrin to obtain a solid content of 35% After adjusting and spray-drying the sample, NKCP-GL 5: Nato bacteria culture concentrate (100 kg) was mixed with 5 kg of Ganoderma lucidum mycelium culture extract concentrate (Brix 30) with indigestible maltodextrin to adjust the solid content to 35%. spray dry sample

²⁾ Average of 3 replicate analysis of 5 lots samples

³⁾ Statistical processing: 0.05% significant difference by Duncan's multiple test (other lowercase letters are significant)

In addition, the present inventors conducted a clinical trial on the thrombolytic ability of the mixed composition according to Example 3, and as a result, both short-term (4,000 U once intake) and long-term (2,000 U once every four weeks) experiment In addition, the effect of remarkably enhanced thrombolysis was confirmed, and it was also verified that there was an anti-arteriosclerosis effect that increased the HDL-cholesterol level of plasma and decreased TG / HDL-cholesterol to improve the anti-arteriosclerosis index.

On the other hand, while the present invention has been shown and described with respect to certain preferred embodiments, the invention is variously modified and modified without departing from the technical features or fields of the invention provided by the claims below It will be apparent to those skilled in the art that such changes can be made.

The present invention is characterized in that it comprises a Bacillus subtilis natto culture and a mushroom culture, to provide a composition that can significantly increase the thrombolytic ability of NATO bacteria by the mushroom culture. Can be.

Claims (5)

Thrombolysis composition comprising a natto bacteria culture and a mushroom culture. [Claim 2] The composition for dissolving thrombus of claim 1, wherein the culture of Nato bacteria is a culture of Bacillus subtilis natto bacteria. According to claim 1, wherein the mushroom culture is a mycelium of Agaricus blazei , Pleurotus ostreatus , Ganoderma lucidum , Lentinus edodes or Lentinus edodes or Coriolus versicolor culture Thrombolysis dissolution composition, characterized in that the culture. The method according to claim 2 or 3, wherein the natto bacteria cultures and mushroom cultures, A composition for dissolving a blood clot, which is included in a weight ratio of 100: 1 to 2. The method according to claim 4, wherein the natto bacteria cultures and mushroom cultures, Thrombolysis dissolving composition, characterized in that the powdered powder powder after concentration.