KR101806130B1 - Antimicrobial and antioxidant composition comprising active components from fermented powder of lactic acid bacteria and herb extract for meat products - Google Patents

Abstract

The present invention relates to a complex antibacterial and antioxidant composition using a herbal extract containing lactic acid fermented powder, rosemary extract and green tea extract. In the present invention, in order to inhibit slime causative bacteria causing malodor, swelling and decay in meat products, a complex composition comprising a fermented lactic acid bacterium produced by Lactobacillus casei subsp. Lambda sp. And a herbal extract having antimicrobial and antioxidant activities at the same time , Which is characterized by long-term antimicrobial activity even at a low concentration and by preventing oxidation and maintaining the leading position of meat products.
The composition according to the present invention can be used for other foods, fermented products, feeds and the like in addition to meat processing for antibacterial and antioxidant use.

Description

TECHNICAL FIELD The present invention relates to an antimicrobial and antioxidant composition comprising a fermented powder of lactic acid bacteria and an extract of herbs as active ingredients,

The present invention relates to an antibacterial composition using fermented lactic acid bacteria and herbal extracts (rosemary, green tea). More specifically, the present invention relates to a composition for inhibiting harmful bacteria causing slime in meat products ≪ / RTI >

With the development of science and civilization and the tendency to pursue convenience, the use of instant foods such as frozen and chilled foods, which emphasize simplicity, has increased, and especially the proportion of meat products such as ham and sausage has increased.

Causes of deterioration during the distribution process of meat products include slime due to microbial growth, color change due to oxidation, and odor. In the case of slime phenomenon, it is the primary or secondary metabolite that occurs in the form of viscous substance outside the cell wall of the microorganism, which is the biggest deterioration factor in the distribution process of meat products. Lactic acid bacteria such as Leuconostoc, Weisella and Lactobacillus have been reported as the main causes of such slime phenomenon. It has been reported that the number of microbial cells tends to increase largely when the air is well processed and packaged during the distribution process. In the case of color change and odor generation, quality deterioration occurs due to oxidation of fat and is a main cause of deterioration of the leading of meat products. Therefore, a safe natural preservative that can simultaneously inhibit the growth and oxidation of microorganisms is indispensable for the safe distribution of increasingly increasing meat products.

A variety of synthetic preservatives are used in meat products to preserve these meat products freshly.

It is known that potassium sorbate, which is the most representative synthetic antimicrobial agent added for the preservation of meat processing products, can cause fake allergy and it can cause urticaria, asthma, rhinitis and hepatotoxicity when overdosed. It is one.

As a synthetic antioxidant, butylated hydroxyl anisole (BHA) and butylated hydroxyl toluene (BHT) are frequently used in meat products. However, the above compounds are a phenolic synthetic antioxidant, and there is a tendency that consumers are increasingly refusing and avoiding them because there are carcinogenic substances.

For the above reasons, it is necessary to develop a natural preservative which can replace the synthetic preservative. Accordingly, many natural materials have been studied.

As a natural antimicrobial material, lactic acid bacteria fermented powder has high antimicrobial activity against lactic acid bacteria such as Leuconostoc, Weisella, and Lactobacillus, which are known to be the main cause of slime, which is a major deterioration phenomenon of meat processing products , It is known. The antimicrobial activity of lactic acid bacteria is already known from existing patents such as antimicrobial peptide materials (registered patent No. 10-0413335), broiler-derived probiotics (registered patent No. 10-1086239), and antimicrobial agents for pharmaceuticals (registered patent No. 10-0483369). In addition, commercially available nisin produced by Lactococcus lactis has now been shown to be safe and used as a preservative in over 50 countries.

As a natural antioxidant, herb extract has been widely used as an antioxidant in the past by using leaves, flowers, roots and stems. The antioxidative activity of the herb was evaluated by the following criteria: antioxidant surface food (registered patent No. 10-0414016); food line maintenance (registered patent No. 10-1205680); antioxidant herbal cosmetic composition (registered patent No. 10-0820010) 10-1222677), and the like. Currently, herbal extracts such as green tea and rosemary are widely used as antioxidants for commercial purposes. It also has the ability to inhibit the growth of microorganisms. When applied to meat processing products, it can improve quality and shelf stability by improving pigment fixation and antibacterial properties.

The above-mentioned conventional techniques are used as a substitute for synthetic preservatives, but their use is limited compared to synthetic preservatives due to their insufficient effectiveness. In addition, there is no report on natural preservatives that can satisfy both antimicrobial activity and antioxidant activity. In particular, there has been no report on antimicrobial and antioxidant inventions for safe preservation of meat products for the purpose of the present invention.

In order to solve the problems of the prior art, if a natural antimicrobial and antioxidant are developed using the synergistic effect of the combined composition comprising the fermented lactic acid bacteria powder and the herbal extract, the composition can be effectively applied to foods.

KR 10-1086239 B1 (2011. 11. 17) KR 10-1205680 B1 (27 Nov. 2012)

Currently, synthetic additives used as antimicrobial and antioxidants in meat products tend to be rejected and avoided by consumers because of safety concerns. Existing natural materials used as a substitute for synthetic preservatives have proved safe, but their use is still limited compared to synthetic preservatives due to their lack of efficacy.

The present inventors intend to develop a composite composition for meat processing having both antimicrobial activity and antioxidative activity, which are natural ingredients of lactic acid bacteria fermented powder, rosemary extract and green tea extract, which are natural materials, and which are not present in the past.

The present invention relates to an antimicrobial composition using fermented lactic acid bacteria and herbal extracts. More particularly, the present invention relates to an antimicrobial composition using lactic acid bacterial fermentation powders and herbal extracts, A food preservative composition capable of preventing spoilage.

The invention is a natural fermented powder and herb-based natural fermented powder that can prevent ingestion of harmful components of synthetic additives commonly used in meat processing products. It is a safe natural preservative to prevent deterioration during the distribution period of meat products. Lt; / RTI >

In the present invention, the antimicrobial activity of a composite composition against a harmful strain producing slime in a meat product was analyzed, and the antimicrobial activity and antioxidative effect were confirmed by applying it to a meat product. The composite composition of the present invention is particularly suitable for meat products, but the application is not limited thereto. The antimicrobial composition of the present invention can also be used for other foods and feeds.

In the following, the present invention will be described in more detail with reference to specific examples. However, it should be understood that the scope of the present invention is not limited by the following embodiments, and that those skilled in the art will readily understand that the technical idea of the present invention and the equivalents of the claims It goes without saying that various modifications and variations are possible.

Experiment 1. Identification of slime causative bacteria in meat processing products

In order to isolate the strain in the present invention, slime generated in domestic meat products (Vienna, ham, bacon) was aseptically collected and the causative bacteria were isolated therefrom. The collected samples were homogenized with sterilized saline (0.85% NaCl) and homogenized. Lactobacilli MRS solid medium (Difco Cat.), Which was a culture medium supplemented with 0.002% of BPB (Bromophenol Blue) No. 288130) and cultured at 30 占 폚 for 24 to 48 hours. From the medium, eight colonies (A to H) having different colony colonies were isolated from colonies formed in blue. The results and physiological characteristics of the isolated strains are shown in Table 1 below. The isolated strains were suspended in 20% glycerol solution, stored at -80 ° C, and cultured at 30 ° C for 24-48 hours after each test.

Identification of slime causative microorganisms in meat processing products division A B C D E F G H Separation source Vienna Vienna ham ham Vienna bacon bacon bacon Gram stain positivity positivity positivity positivity positivity positivity positivity positivity Catalase voice voice voice voice voice voice voice voice shape coccus coccus coccus coccus Bacillus Bacillus Bacillus Bacillus Identification result Leuconostoc sp. Lactobacillus sp. Weissella sp.

Example 1 Preparation of Fermentation Powder of Lactic Acid Bacteria

The present inventors selected microorganisms having the best antimicrobial activity against the perishable microorganisms from the 48 week microorganisms obtained from infant feces, crude oil and kimchi, and analyzed the microbiological properties of the microorganisms. As a result, it was confirmed that it belongs to Lactobacillus genus. The strain Lactobacillus casei subspecies ramno Versus eseuenti blood 01: named (Lactobacillus casei subsp rhamnosus SNTP01, Accession No. KACC 91057), which was used in the present invention. The Lactobacillus casei subsp. Lambusus strains 01 were cultured in Lactobacillus MRS medium at 37 ° C for 24 hours and centrifuged at 13,000 rpm for 15 minutes to remove the cells. The cells were concentrated to a concentration of 0.01 to 0.1% by weight and dried. The dried powder was used as a final lactobacillus fermentation powder sample.

Example 2: Preparation of rosemary extract

To produce rosemary extract having excellent antioxidant activity, the present inventors extracted rosemary leaves by pulverizing 0.5 to 5 mm of leaves and adding 5 to 20 times of 50 to 100% of alcohol to the pulverized leaves. The alcohol-added extract was concentrated to dryness under reduced pressure of 0.2 to 0.5% based on the weight of the leaves, and the dried powder was used as a final rosemary extract sample.

Example 3. Preparation of green tea extract

In order to prepare a green tea extract having excellent antioxidant activity, the present inventors extracted green tea leaves by pulverizing 0.5 to 5 mm of leaves and adding 5 to 20 times of 50 to 100% of alcohol to the pulverized leaves. The alcohol-added extract was concentrated to a concentration of 0.2 ~ 0.5% by weight of the leaves and dried. The dried powder was used as a final green tea extract sample.

Examples 4 to 6. Preparation of Composite Composition

Composite compositions were prepared using the fermented lactic acid bacteria powder, rosemary extract, and green tea extract prepared in Examples 1, 2, and 3, and the compounding ratios are shown in Table 2 below.

Composition ratio of composite composition division Example 1 (%) Example 2 (%) Example 3 (%) Example 4 20 40 40 Example 5 30 40 30 Example 6 50 30 20

Experiment 2. Identification of antibacterial range of composite composition

In order to confirm the antimicrobial activity against the slime causative microorganisms of the meat products of the extracts and the composite compositions prepared according to Examples 1 to 6, the antibacterial activity was tested on eight kinds of isolated lactic acid bacteria isolated in Experiment 1. [ The antimicrobial activity was measured by the broth dilution method according to CLSI (The Clinical and Laboratory Standards Institute). The culture broth containing 8 different lactic acid bacteria was cultivated in Lactobacillus MRS medium at 30 ° C for 24 hours with a certain concentration of the antimicrobial sample and cultured in a 96-well plate containing Lactobacillus MRS medium in a stepwise two- fold dilution method. The antimicrobial activity of the samples prepared by absorbance at 600 nm was investigated. The minimum inhibitory concentration (MIC), the concentration at which the growth of the test strain was inhibited by 90%, was indicated. Potassium sorbate was used as a control. All the experiments were repeated three times to prepare an average value, and the results are shown in Table 3 below.

The antimicrobial effect (MIC) of the composite composition on 8 species of slime causative bacteria Meat-harvesting bacteria The prepared composite composition (ppm) Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 Potassium sorbate A 78 625 625 312 250 125 780 B 126 1,250 1,250 630 312 156 1,260 C 156 1,000 1,250 625 500 156 1,260 D 126 625 500 500 250 156 1,260 E 126 625 625 625 250 126 1,260 F 126 625 1250 500 312 156 1,260 G 78 312 500 250 126 78 780 H 126 625 312 500 312 250 1,260

As a result, the fermented lactic acid bacteria showed the best antimicrobial activity alone, and the composition of Example 6 (fermented lactic acid bacteria powder 50, rosemary 30, green tea 20 weight ratio) showed the most antimicrobial activity.

Particularly, when the composite composition was prepared, the antimicrobial activity against weight was higher than that obtained by using alone. In Example 6, the antimicrobial activity was 1.5 to 2 times better than that of the lactic acid bacteria fermented powder, rosemary extract and green tea extract, respectively. This is a result of the synergistic effect between the lactic acid bacteria fermentation powder and the herbal extract which have not been conventionally known.

Experiment 3. Confirmation of Antioxidative Activity of Composite Composition

DPPH (1,1-diphenyl-2-picryl hydrazyl) anion scavenging activity was evaluated in order to evaluate the antioxidative activity of the extracts and the composite compositions prepared in Examples 1 to 6 above. For DPPH anion scavenging activity, 380 of 2 × 10 -4 M DPPH solution in 99.5% ethanol was added to sample 20 diluted to various concentrations, and the mixture was reacted at 37 ° C. for 30 minutes. Then, a microplate reader (Asys Hitech , Expert 96, Asys Co., Austria). Vitamin C (Vit. C, Sigma Co., USA) was used as a control. The DPPH free radical scavenging activity was expressed as the percentage of sample addition and non-addition, and the IC ₅₀ was calculated as the concentration exhibiting 50% scavenging ability. The final result was expressed as the average of three measurements.

ASA (%) = (C-S) / Cx100

C: Absorbance at the time of addition of solvent control DMSO

S: absorbance at the time of sample addition

Antioxidant activity value of the composite composition DPPH
Scatters The resulting composite composition (IC ₅₀ , 占 퐂 / ml) Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 Vit. C 73.33 10.29 13.75 7.21 9.98 16.24 13.29

As shown in the above results, the rosemary extract showed the best antioxidative activity alone, and the composition of Example 4 (fermented lactic acid bacteria powder 20, rosemary 40, green tea 40 weight ratio) showed the best antioxidative activity.

Particularly, in Example 4, which is a composite composition, an excellent antioxidative activity was observed 1.4 to 2 times or more as compared with the result of using alone. As in Experiment 2, the antioxidant activity was enhanced by the synergistic effect between the fermented powder of lactic acid bacteria and the herb extract.

When the above results are summarized, the antimicrobial activity of the composite composition was excellent in the order of Examples 6, 5 and 4, and antioxidant activity was high in the order of Examples 4, 5 and 6.

Examples 7 to 11 and Comparative Examples 1 to 2: Manufacture of ham with a composite composition

Hams supplemented with the extracts and the composite compositions prepared in Examples 1 to 6 were prepared. The blending ratios used in the preparation are shown in Table 5. In order to confirm the synergistic effect of the composition upon application of the ham, hams supplemented with the extract prepared in Examples 1 and 2 were also prepared. In Comparative Example 1, potassium sorbate, vitamin C in Comparative Example 2 and preservative in Comparative Example 3 were not added Ham. Fat-removed pork was used, prepared according to the blend ratio, tumbled at 4 and placed in a cold place for 6 hours. After drying for 50 to 40 minutes, it was smoked for 60 to 30 minutes, then heat-treated, cooled, and vacuum-packed.

Mixing ratio used in the manufacture of meat products Name of raw material ratio (%) Example 7 Example 8 Example 9 Example 10 Example 11 Comparative Example 1 Comparative Example 2 Comparative Example 3 Pork 96 96 96 96 96 96 96.3 96.3 Purified salt 1.8 1.8 1.8 1.8 1.8 1.8 1.8 1.8 Sodium L-glutamate 0.3 0.3 0.3 0.3 0.3 0.3 0.3 0.3 Powder egg white 0.4 0.4 0.4 0.4 0.4 0.4 0.4 0.4 Carrageenan 0.6 0.6 0.6 0.6 0.6 0.6 0.6 0.6 Sodium nitrite 0.02 0.02 0.02 0.02 0.02 0.02 0.02 0.02 Pigment 0.58 0.58 0.58 0.58 0.58 0.58 0.58 0.58 Example 1 0.3 0 0 0 0 0 0 0 Example 2 0 0.3 0 0 0 0 0 0 Example 4 0 0 0.3 0 0 0 0 0 Example 5 0 0 0 0.3 0 0 0 0 Example 6 0 0 0 0 0.3 0 0 0 Potassium sorbate 0 0 0 0 0 0.3 0 0 vitamin C 0 0 0 0 0 0 0.3 0 Sum 100 100 100 100 100 100 100 100

Experiment 4. Confirmation of the preservation effect of the ham on the mixed composition treatment

The spread plate method was used to evaluate the inhibition of lactic acid bacterial growth of the ham prepared above. The samples were stored at 10 ° C and sampled at intervals of 3 to 4 days. The collected samples were homogenized with sterilized saline (0.85% NaCl), homogenized, and then inoculated 100 ml into Lactobacilli MRS solid medium, a culture medium for lactic acid bacteria containing 0.002% BPB by a 10-fold dilution method, And cultured at 30 DEG C for 48 hours. The number of lactic acid bacteria in the solid medium was examined, and the results are shown in Table 6.

Inhibition of Lactic Acid Bacteria Growth in Meat Products by Addition of Composite Composition Name of sample division Number of lactic acid bacteria (CFU / g) 3 day 7 day 11 day 16 day 21 day 26 day 30 day ham Example 7 8x10 ¹ 6x10 ¹ 3x10 ¹ <10 <10 <10 <10 Example 8 2x10 ² 8x10 ² 6x10 ³ 7x10 ⁵ 10 ⁶ < 10 ⁶ < 10 ⁶ < Example 9 1x10 ² 4x10 ¹ <10 <10 <10 <10 <10 Example 10 6x10 ¹ 2x10 ¹ <10 <10 <10 <10 <10 Example 11 2x10 ¹ <10 <10 <10 <10 <10 <10 Comparative Example 1 2x10 ¹ <10 <10 <10 <10 <10 <10 Comparative Example 3 4x10 ² 1x10 ⁵ 1x10 ⁶ 10 ⁶ < 10 ⁶ < 10 ⁶ < 10 ⁶ <

In all the results of the ham containing the composite composition, high proliferation inhibition results were observed compared to Comparative Example 3 which was not treated. In Comparative Example 3, 1 x 10 ⁶ CFU / g of lactic acid bacteria was observed on the 11th day, whereas in the meat products to which the composite composition was added, the lactic acid bacterial growth was inhibited to ¹ x 10 ¹ CFU / g or less by 30 days. This result is equivalent to that of Comparative Example 1 in which potassium sorbate is contained and it is confirmed that the preservation effect is excellent.

Also, in Example 11, which is a composite composition, the bacteria were killed more rapidly than Example 7 in which the lactic acid bacteria fermented powder having the best antimicrobial activity was added alone in Experiment 2. This was more effective when the composite composition was applied to a meat product such as ham than the result of the simple antibacterial activity test as in Experiment 2, by exhibiting more effective antibacterial effect.

Experiment 5. Evaluation of Oxidation Stability in Meat Products by Composite Composition Treatment

To evaluate the oxidation stability of the meat products prepared in Examples 7 to 11, fatty acid pigments were measured by 2-thiobarbituric acid reactive substances (TBARS) method. To 20 g of the sample, 50 ml of 20% trichloroacetic acid (in 2 M phosphate) reagent was added to homogeneity. The mixture was adjusted to 100 ml with distilled water and filtered. To the filtrate was added the same amount of 2-thiobarbituric acid (TBA, in water) and shaken for 15 hours. The absorbance was measured at 530 nm. The samples were collected at intervals of 3 to 4 days, and TBARS was measured. The results are shown in Table 7.

Fatty Acids in Meat Products by Addition of Composite Composition Name of sample division Fatty acids according to the number of days of storage (mg MDA / kg) 3 day 7 day 10 day 13 day ham Example 7 0.421 0.442 0.469 0.531 Example 8 0.397 0.431 0.451 0.472 Example 9 0.385 0.416 0.435 0.457 Example 10 0.392 0.432 0.451 0.480 Example 11 0.413 0.442 0.463 0.501 Comparative Example 2 0.402 0.428 0.449 0.465 Comparative Example 3 0.425 0.457 0.482 0.552

As a result of the addition of the complex composition, the TBARS as a whole increased with the storage period, and the oxidation inhibition was significantly inhibited in the example in which the composite composition was added, as compared with Comparative Example 3 in which the composition was not treated. In Example 9, the TBARS value was 0.03 or lower from the 3rd day from the third comparative example, from the 10th day in the example 10, and from the thirteenth day in the example 11. This indicates that the highest antioxidative activity is shown in Example 9, which exhibits the fastest significant difference and inhibits oxidation, and that the antioxidative activity is high in the order of Example 10 and Example 11. [ Overall, it was confirmed that the addition of the combined extract effectively delayed oxidation of fat in the ham. The above results confirmed that the oxidation inhibiting effect was superior to that of Comparative Example 2 in which vitamin C was contained.

Also, in the case of Example 9 and Example 10 of the composite composition, it was observed that the fat oxidation was slower than in Example 8 in which the rosemary extract alone, in which the highest antioxidative activity was observed, was observed from Experiment 3 described above. This indicates that when the composite composition is applied to a meat product such as ham, the antioxidant activity is enhanced as compared with the individual extract.

According to the results of Experiment 4, bacteria were killed more rapidly than in Examples 7 and 8, which were added alone in all the Ham treatments to which the complex composition was added in Examples 9 to 11. Also, according to the results of Experiment 5, the antioxidative activity was superior to that of Example 7 and Example 8, in which the Ham treatment groups to which the composite composition was added were added singly.

Therefore, when the above results are comprehensively judged, it was confirmed that Examples 9 and 10 of the composite composition are the most excellent compositions capable of simultaneously securing high antimicrobial and antioxidative activities when applied to meat processing.

Claims (3)

A composition having antimicrobial and antioxidative activity for pork processing products,
Characterized in that which consists of the fermented grains of KACC 91057), b) rosemary extract, and c) green tea extract: The hybrid composition is a) Lactobacillus casei subspecies ramno Versus eseuenti blood 01 (Lactobacillus casei subsp rhamnosus SNTP01, Accession No. Of preserving ingredients for leading-edge pork processing products
The method according to claim 1,
Wherein the preservative composition is a) 20 to 30 parts by weight of a fermented powder of Lactobacillus casei subsp. &Lt; RTI ID = 0.0 &gt; lambosus &lt; / RTI &gt; Sustifie 01, b) 40 parts by weight of rosemary extract, and c) 30 to 40 parts by weight of green tea extract. Preservative composition
3. The method of claim 2,
Wherein said preservative composition is added to food or animal feed.