KR101391284B1 - Novel lactobacillus plantarum to-2100, fermentation method of mulberry leaves using thereby, fermented materials thereby and food composition comprising the fermented materials - Google Patents

Abstract

The present invention is Lactobacillus Room Planta TO-2100 (Lactobacillus plantarum TO-2100, accession number KFCC 11537P) and fermentation of the mulberry leaf method using the same, and thus of mulberry leaves fermentation and relates to a food composition comprising the same, and more particularly, to a novel fiber-degradable lactic acid bacteria of Lactobacillus Planta room TO- 2100 ( Lactobacillus plantarum TO-2100, accession no. KFCC 11537P), which has an inhibitory activity on α-glycosidase, which is a carbohydrate metabolizing enzyme, and piperazine alkaloids A Lactobacillus planta TO-2100, a fermentation method of mulberry leaf using the same, a fermented product of mulberry leaf and a food composition containing the same.

Description

TECHNICAL FIELD [0001] The present invention relates to a novel lactobacillus planta TO-2100 and a fermentation method of mulberry leaf using the same, and a fermented product of mulberry leaf and a food composition containing the same. TECHNICAL FIELD [0001] The present invention relates to novel lactobacillus plantarum TO- COMPOSITION COMPRISING THE FERMENTED MATERIALS}

The present invention is Lactobacillus Room Planta TO-2100 (Lactobacillus plantarum TO- 2100, Accession No. KFCC 11537P), a fermented mulberry leaf and a food composition containing the fermented mulberry leaf. More particularly, the present invention relates to a novel fiber-degrading lactic acid bacterium Lactobacillus plantarum TO-2100 ( Lactobacillus plantarum TO-2100, accession number KFCC 11537P), the content of nitrogen-containing compounds and piperidine alkaloids having inhibitory activity of α-glycosidase, which is a carbohydrate metabolizing enzyme, To a fermented mulberry leaf using the same, and a food composition containing the fermented mulberry leaf.

According to Mulberry leaves, Maternal Maternity Mushroom, Primrose Mushroom, Japanese Mushroom, Mushroom, and Dongbu Health, it is said that Mulberry leaves are caused by diseases such as swine fever, edema, diabetes, Etc. have been reported as medicinal plants. Mulberry leaves contain more minerals than green tea and contain flavonoids and steroids such as rutin, quercetin, quercitrin and isoquercitrin, which are functional ingredients, It contains amino acids and vitamins. In addition, the inhibitory neurotransmitter of the central nervous system is relatively abundant in γ-aminobutyric acid (GABA), which plays an important role in suppressing the increase of blood pressure, controlling appetite and satiety, (1-deoxynojirimycin, 1-DNJ) and nitrogen-containing saccharide (N-glycosidase), which inhibit the activity of glucosidase, containing sugar in a large amount. It is a natural material that is most notable in the treatment and prevention of diabetes, which is expected to occur anywhere in the world and to increase to 4.4% of the world's population in 2030.

Medicinal plants have been used in the form of biological and dry powders for therapeutic purposes. Water and organic solvent extraction methods have been developed to solve problems such as overage and short shelf life. However, such a method has problems such as a change in the extracting substance depending on the polarity of the extraction solvent and a decrease in the yield due to the density of the plant cell wall.

To overcome these disadvantages, fermentation extraction using microorganisms has attracted attention. The microorganisms used in the fermentation process are bacteria and fungi capable of decomposing fibrous cells of plant cells and yeasts producing alcohol. This method of fermentation has the advantage of increasing extraction yield by gradually magnetizing plant cell walls, gradually increasing the alcohol content, facilitating the extraction of useful components, and converting unnecessary sugars present in plants into useful substances have.

Commonly used submerged fermentation has several disadvantages, including complicated equipment and waste after fermentation, while solid-state fermentation uses low-cost raw materials such as inexpensive equipment and agricultural wastes It is an environmentally friendly way to produce high-efficiency products and not to produce waste.

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made keeping in mind the above problems occurring in the prior art, and it is an object of the present invention to provide a pharmaceutical composition for preventing or ameliorating a disease caused by 1-deoxynojirimycin (1-DNJ) having an alpha-glucosidase- Ferritin alkaloids are easily extracted under low acidic conditions, it is possible to produce a novel fiber-degradable lactic acid bacterium Lactobacillus planta TO-2100 ( Lactobacillus < RTI ID = 0.0 > plantarum TO- 2100, accession number KFCC 11537P).

The present invention is a Lactobacillus Planta room TO-2100 (Lactobacillus of the new fiber-degradable lactic acid bacteria plantarum TO- 2100, Accession No. KFCC 11537P), it is possible to increase the content of nitrogen-containing compounds and piperidine alkaloids having an inhibitory activity of α-glycosidase, which is a carbohydrate metabolizing enzyme It is another object of the present invention to provide a fermentation method of mulberry leaves using a novel Lactobacillus plantarum TO-2100.

In addition, the present invention relates to novel Lactobacillus Planta room TO-2100 (Lactobacillus plantarum TO -2100 , accession number KFCC 11537P) is fermented by a fermentation method of the mulberry leaf in the alpha-glucosidase (α-glycosidase) carbohydrate-metabolizing enzymes with And a fermented mulberry leaf having an increased content of piperidine alkaloids and the like.

In addition, the present invention relates to novel Lactobacillus Planta room TO-2100 (Lactobacillus plantarum TO-2100 , Accession No. KFCC 11537P), and the content of nitrogen-containing compounds and piperidine alkaloids having an inhibitory activity of α-glycosidase, which is a saccharide metabolizing enzyme, It is another object of the present invention to provide a food composition comprising an increased fermented mulberry leaf.

The problems to be solved by the present invention are not limited to those mentioned above, and other problems to be solved can be clearly understood by those skilled in the art from the following description.

In order to achieve the above object, the present invention provides a novel Lactobacillus plantarum TO-2100 TO- 2100 , accession number KFCC 11537P).

The present invention also relates to a Lactobacillus plantarum TO-2100 ( Lactobacillus < RTI ID = 0.0 > plantarum TO- 2100 , accession number KFCC 11537P).

In addition, the fermentation process of the mulberry leaf in accordance with the invention is characterized in using the novel Lactobacillus Planta room TO-2100 (Lactobacillus plantarum TO -2100 , accession number KFCC 11537P) fermenting the mulberry leaves.

The method of fermenting mulberry leaves according to the present invention is characterized by solid fermentation.

Also, the fermentation method of mulberry leaves according to the present invention is characterized in that the content of 1-Deoxynojirimycin and Piperidine Alkaloid contained in the mulberry leaves is increased.

The mulberry leaf fermentation method according to the present invention is characterized in that the initial moisture content of fermentation is 20%, the fermentation temperature is 30 to 35 ° C, and the fermentation relative humidity is 60 to 70%.

Meanwhile, the mulberry leaf fermented according to the present invention is characterized in that it is fermented by a fermentation method of mulberry leaf using a novel Lactobacillus plantarum TO-2100 (Accession No. KFCC 11537P).

Meanwhile, the food composition according to the present invention is characterized by including the fermented mulberry leaf.

According to the present invention having the above-described constitution, 1-deoxynojirimycin (1-DNJ) having an alpha-glucosidase inhibitory activity as a saccharide metabolizing enzyme is reacted with piperidine alkaloid In view of the fact that they are easily extracted in an acidic condition, a new fiber-decomposing lactic acid bacterium Lactobacillus plantata TO-2100 ( Lactobacillus plantarum TO- 2100, Accession No. KFCC 11537P), a fermentation method of mulberry leaves using the same, a fermented mulberry leaf thereof, and a food composition containing the same, thereby developing new microorganism resources and manufacturing and developing functional foods, Treatment, and bioprocess engineering.

The effects attained by the present invention are not limited to those mentioned above, and other effects not mentioned can be clearly understood by those skilled in the art from the following description.

Figure 1 of the present invention Lactobacillus bacteria Planta room TO-2100 (Lactobacillus plantarum TO-2100) by 16S rDNA sequencing analysis.
Figure 2 is a photograph of the Lactobacillus planta TO-2100 ( Lactobacillus < RTI ID = 0.0 > Plantarum TO-2100) phylogenetic tree.
Figure 3 is a photograph of the Lactobacillus plantarum TO-2100 ( Lactobacillus < RTI ID = 0.0 > plantarum TO-2100) TLC pattern of piperidine alkaloids extracted from the fermented mulberry leaf powder.
Fig. 4 shows the results of comparing the inhibitory activity of alpha-glucosidase.

Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the following description. However, the present invention is not limited to the embodiments described herein but may be embodied in other forms. Rather, the embodiments disclosed herein are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art. Like numbers refer to like elements throughout.

Figure 1 of the present invention Lactobacillus bacteria Planta room TO-2100 (Lactobacillus plantarum TO-2100) . Fig. 2 shows the results of 16s rDNA sequencing analysis of the Lactobacillus planta TO-2100 ( Lactobacillus plantarum TO -2100). Fig. 3 is a phylogenetic tree of Lactobacillus planta TO-2100 ( Lactobacillus plantarum TO-2100), and FIG. 4 is a TLC pattern of the alpha-glucosidase inhibitory activity of the piperidine alkaloid extracted from the mulberry leaf powder fermented by solid fermentation.

Example  1. Isolation of Lactic Acid Bacteria Having Fiber-degrading Activity

In order to isolate lactic acid bacteria, the present invention was collected from traditional Korean leeks in Taebaek, Kangwon, Iksan, Jeonbuk, and Geumjeong, Busan. The collected nuruk were suspended in physiological saline, filtered, and diluted to an appropriate magnification. The diluted solution was cultured on MRS (Lactobacilli MRS) medium for lactic acid bacteria and cultured at 37 ° C for 24 hours. The colonies formed after cultivation were first inoculated into MRS medium supplemented with 2% CaCO ₃ to select 15 strains whose size of transparent ring was 5 mm or more from the cells. Lactobacilli MRS agar medium for lactic acid bacteria supplemented with 10 g / L carboxymethyl cellulose except for dextrose and ammonium citrate and 15 kinds of selected primary strains And cultured at 37 DEG C for 48 hours. After culturing, staining with 2% I ₂ , 3% KI in Gram's Iodine solution dissolved in 70% ethanol and finally forming a yellow transparent ring were finally selected.

To isolate the lactic acid bacteria having the ability to decompose the fiber for lactic acid fermentation of mulberry leaves, 15 strains having a transparent ring size of 5 mm or more in the MRS medium supplemented with 2% CaCO ₃ were added to MRS medium containing carboxymethyl cellulose (CMC) When Gram's Iodine was stained, the strain with the largest size of yellow transparent ring was finally selected and named TO-2100.

Example  2. CTR -2100 Identification

(1) morphology and culture characteristics

The size and shape of the strains were observed by Gram staining according to the method of Gerhardt et al. (1994) and observed with an optical microscope (Nicon, FK-IIA, Japan). The exact appearance of the isolates was 2.0% PTA (phosphotungstinic acid ) With negative staining and observed with an electron microscope (Hitachi S-4800, Japan).

The selected TO-2100 strain had a colony shape in the acidic plate medium (pH 3.0), a round color, a bright milky white color, and a convex and smooth state on the surface, indicating the lactic acid-specific characteristic of lactic acid bacteria.

(2) Physiological characteristics

The physiological characteristics of the strain include casein resolving ability, starch resolving ability, gelatin liquifying power, sugar fermentation ability, indole producing ability, nitrate reducing power, catalase and oxidase oxidase production was investigated according to the microbiology laboratory manual.

The results of examining physiological characteristics of TO-2100 are shown in Table 1. Growth pH ranged from 2.0 to 9.0 and growth temperature ranged from 15 to 45 ℃. In addition, it was possible to grow up to 15% salt concentration, catalase positive, oxidase positive, starch, casein hydrolyzed, and cellulose hydrolysis was positive. It also had indole production ability and produced NH ₃ from arginine and peptone. No growth was observed in neutral nutrient medium.

(3) Cellular fatty acid analysis

The whole cell fatty acid composition of the strain was analyzed using gas chromatography (GC, HP 6890). The standard fatty acid was obtained from Hewlett-Packard (HP), and the culture was carried out using Sabouraud's agar (Difco, USA) medium at 30 ° C for 24 hours After culturing, the cultured cells were taken in a test tube and 1 ml of a saponification reagent (45 g sodium hydroxide (NaOH), 150 methanol (CH ₃ OH, distilled water, distilled water) RTI ID = 0.0 > C < / RTI >

The saponified cells were methylated by adding 2 ml of a 2.6: 1 mixture of 6 N HCl and methanol, followed by methylation at 80 ° C for 10 minutes, and extraction of hexane and methyl tert-butyl ether 1.25 ml of a 1: 1 (v / v) mixture was added, and the mixture was slowly shaken for 10 minutes at room temperature. After washing with 5.4% NaCl, the sample was used as an analytical sample. Analytical samples were analyzed by GC and analyzed by Sherlock program, a MIDI strain identification program.

As a result of examining the cellular fatty acid component of the TO-2100, the contents of the main components were 18: 1 and 16: 0 normal, respectively, and the contents were 51.63% and 16.17%, respectively (Table 2). Lactobacillus and Bifidobacterium The 18: 1 normal, the major fatty acid component of Lactobacillus and Bifidobacterium bacteria, varies from 25 to 55% and 17 to 40%, 16: 0 normal to 21 to 35% and 15 to 38%, respectively, Which is consistent with the report of Veerkamp (1971). According to this report, the content of 18: 0 was relatively high in Bifidobacterium spp. (4 ~ 22%), while in Lactobacillus spp. (0.5 ~ 4.0%). Therefore, the TO-2100 strain was estimated to be lactic acid bacteria of the genus Lactobacillus .

(4) 16s rDNA sequencing

16S rDNA gene analysis was performed by PCR (polymerase chain reaction, polymerase chain reaction) using Wizard Genomic DNA Prep Kit (Promega). At this time, the reaction conditions were denaturation at 94 ° C for 1 minute, annealing at 60 ° C for 1 minute and 30 seconds, extension at 72 ° C for 1 minute, and this operation was repeated for 30 cycles in total. The primers used were 27F (5'AGAGTTTGATCMTGGCTCAG) and 1492r (5'TACGGYTACCTTGTTACGACTT).

The amplified fragment (16S rDNA region) was inserted into pGEM T Easy vector (Promega) and transformed by a general molecular cloning method using E. coli DH5 as a host cell (small scale preparation) of plasmid using Wizard Plus SV miniprep DNA purification system and automated DNA sequencer (ABI 3700, Applied Biosystems Inc.) was used. Figure 1 shows the results of 16s rDNA sequencing analysis of the TO-2100 strain.

Alignment was obtained by aligning the sequence with reference to the rRNA secondary structure using Clustal X (ver. 1.8) software, and then calculating the simmilarity value. Also, we compared the nucleotide sequence of the genebank with the blast search (advanced blast search). As a result of 16s rDNA gene alignment from the DNA of the sample, the species of the sample was Lactobacillus plantarum The National Institutes of Biotechnology Information (NCBI) National Center for Biotechnology Information (NCBI), the most similar species in the genbank, has been identified as Lactobacillus plantarum > gi | 157907500 | dbj | AB362768.1 | , And 99.67% (1526/1531), respectively. The Lactobacillus planta TO-2100 ( Lactobacillus plantarum TO- 2100 ). A phylogenetic tree was created by the Neighbor-joining (NJ) method and the results are shown in FIG.

The novel Lactobacillus plantarum TO-2100 of the present invention was deposited at the Korea Culture Center for Microorganisms (KCCM) on Jul. 07, 2012, under the name 'KFCC 11537P '.

Example  3. Lactobacillus Flora Room CTR -2100 Lactic Acid Generation  And fibrotic enzyme activity measurement

(1) Lactic acid production ability

The lactic acid production ability of the isolates was measured by pH and titratable acidity. The pH of the isolate was measured by pH meter (sp-701, Suntex Inc. Korea) after shaking at 25 rpm and 150 rpm for 48 h. The lactic acid production rate was measured by Food analysis (Nielsen, 2003) After the titration with 0.1 N NaOH solution until the pH reached 8.3, the amount of 0.1 N NaOH was converted into the lactic acid content (%) by the following equation.

(2) Measurement of Carboxymethyl Cellulose (CMCase) activity

To determine Cx or β-1.4 glucanase activity, 100 ml of 1% carboxymethyl cellulose (CMC), 10 ml of 0.5 mol citrate buffer (pH 4.8) and 1 ml of 1% merthiolate were added according to the method of Mandel and Weber Was diluted with 1 L of distilled water and used as a substrate. After culturing, the cells were removed by centrifugation, and 0.5 ml of the supernatant was added to 0.5 ml of the substrate solution, followed by reaction for 50 and 30 minutes. After the reaction, the reaction was stopped by adding 3 ml of a DNS reagent (Miller, 1959). The reaction was stopped for 5 minutes in boiling water and then colorimetrically measured at 450 nm. The amount of reducing sugar (D-glucose) was calculated from a standard curve prepared beforehand. 1 unit was the amount of enzyme producing 1 mol D-glucose for 1 min.

(3) Filter Paper (FPase) measurement

The filter paper hydrate activity was measured by the method of Whatman No. (1987) according to the method of Ghose (1987). 1 filter paper as a substrate. The filter paper was cut into 16 cm strips (50 mg), and 1 ml of 0.05 mol citrate (pH 4.8) was added and reacted at 50 for 1 hour. The reaction was stopped by addition of 3 ml of DNS reagent and allowed to stand for 5 minutes in boiling water The amount of reducing sugar produced was colorimetrically determined at 450 nm, and the amount of reducing sugar (D-glucose) was converted from a standard curve. 1 unit was the amount of enzyme producing 1 mol D-glucose for 1 min.

(4) Results

The lactic acid production ability and the proteolytic enzyme activity of the isolated strain Lactobacillus plantarum TO-2100 were measured, and the pH was 3.2 and the lactic acid production was 0.68%. The activities of FPase (filter paper cellulase) and CMCase (carboxymethyl cellulase) were 0.35 and 40.1 units, respectively (Table 3).

Example  4. Solid fermentation ( Solid - state fermentation ) Of piperidine of mulberry leaves Alkaloid  Content change

(1) Test method

1) Influence of initial water content

The initial moisture content was 20 ~ 40% (W / V) of sterilized distilled water in the dried mulberry leaf powder pasteurized at 70 ℃ for 24 hours. At this time, the moisture content was measured using an infrared moisture meter. To the mulberry leaf powder adjusted for each initial moisture concentration, the novel Lactobacillus planta TO-2100 ( Lactobacillus plantarum The content of nitrogen compounds and piperidine alkaloids was compared with that of non-fermented mulberry leaves by inoculating 3 ml (10 ⁸ / ml) of the culture broth of TO-2100) and culturing at 30 ° C for 72 hours.

2) Influence of fermentation temperature

To the mulberry leaf powder having a moisture content of 20% treated aseptically, the novel lactobacillus planta TO-2100 ( Lactobacillus plantarum TO-2100) culture broth (3 ml (10 ⁸ / ml)) was inoculated and cultured at 25 to 40 ° C for 72 hours to determine the contents of nitrogen compounds and piperidine alkaloids.

3) Influence of relative humidity

The influence of the relative humidity was such that the mulberry leaf powder having a moisture content of 20% was added to the novel Lactobacillus planta TO-2100 ( Lactobacillus plantarum TO-2100) was inoculated with 3 ml (10 ⁸ / ml) of the culture broth and incubated for 72 hours at 30 ° C with a relative humidity of 50 to 80% using a thermo-hygrostat. The content of nitrogen compounds and piperidine alkaloids Respectively.

4) Determination of nitrogen compound, piperidine alkaloid content and 1-deoxynojirimycin (1-DNJ)

Piperidine alkaloids were extracted by modifying the method of Ahmad et al. (2002). A 20-fold amount of 0.5 mol HCl solution was added to the fermented mulberry leaf powder and extracted at 80 ° C for 5 hours. The extract was filtered under reduced pressure to remove the residue. 4 mol of NH ₄ OH solution was added to the filtrate to adjust the pH to 12 and left at room temperature for 2 hours. The resulting precipitate was measured to obtain a basic nitrogen compound content. The precipitate was redissolved in distilled water and then fractionated with CHCl ₃ . CHCl ₃ The fractions were concentrated under reduced pressure, dried at 70 DEG C for 12 hours, and the content thereof was measured to determine the content of piperidine alkaloids. Dry fraction 1 g / ㎖ of the sample solution 10 ℓ TLC (thin-layer chromatography ) plate (silica gel 60 F254, Merck Co.) dropwise (spotting), and the _{CHCl 3: NH 4 OH: MeOH} (30: 1: 1 ) ≪ / RTI > solvent system. The spot development was sprayed with iodine vapor and compared with the standard materials of 1-deoxynojirimycin (1-DNJ). 1-deoxynojirimycin (1-DNJ) was separated and its content was measured by performing preparative TLC (PTLC) in the same solvent system.

(2) Results

1) Influence of initial water content

Sterilized distilled water was added to pasteurized mulberry leaf powder to adjust the initial moisture concentration to 20 to 40% (w / v), and the novel Lactobacillus plantarum TO-2100 ( Lactobacillus plantarum TO-2100) was inoculated with 3 ml (10 ⁸ / ml) of the culture broth and cultured at 30 ° C for 72 hours. As a result, the content of nitrogen compounds and piperidine alkaloids was measured. The content of nitrogen compounds and piperidine alkaloids was 4.23 and 5.73 times higher than non-fermented extract (non-fermented in Table 4), which was 13.21 g and 2.29 g, respectively, per 100 g of mulberry leaf powder, 3.12 and 0.40 g Respectively. Also, the increase of the initial moisture content decreased the possibility of contamination of germs and fermentation rate.

2) Influence of fermentation temperature

To the mulberry leaf powder whose initial moisture content was adjusted to 20%, a novel Lactobacillus planta TO-2100 ( Lactobacillus plantarum The non-fermented extract (non-fermented in Table 5) was prepared by inoculating 3 ml (10 ⁸ / ml) of the culture broth of TO-2100) and culturing it at 25 to 40 ° C for 72 hours and measuring the contents of nitrogen compounds and piperidine alkaloids. '), The yield was the highest at 30 ~ 35 ℃ and the yield decreased at temperatures above 40 ℃ (Table 5).

3) Influence of relative humidity

To investigate the effect of relative humidity during the fermentation process, the mulberry leaf powder whose initial moisture content was adjusted to 20% was added to the novel lactobacillus planta TO-2100 ( Lactobacillus plantarum TO-2100) was inoculated with 3 ml (10 ⁸ / ml) of the culture broth, and the relative humidity was adjusted to 50 to 80% at 30 ° C using a thermo-hygrostat. After culturing for 72 hours, the nitrogen compound and piperidine alkaloid content Respectively.

As shown in Table 6, the contents of nitrogen compounds and piperidine alkaloids in the relative humidity of 60 to 70% were 16.54 to 16.41 g and 2.86 to 2.82 g per 100 g of mulberry leaf powder, respectively, and the contents of 3.12 and 0.40 g 5.30 ~ 5.26 and 7.15 ~ 7.05 times higher than that of the fermented extract ('Non-fermented' in Table 6), indicating that the solid fermentation method is effective for increasing the functional ingredient. In addition, the decrease in the amount of water produced when the relative humidity is increased is considered to inhibit the growth of microorganisms due to the absence of aeration due to powder aggregation.

4) Content of 1-deoxynojirimycin, 1-DNJ

The novel Lactobacillus plantarum TO-2100 ( Lactobacillus < RTI ID = 0.0 > plantarum TO-2100). The TLC pattern of the piperidine alkaloid extracted from the mulberry leaf powder fermented by solid fermentation is shown in FIG. In Fig. 3, A represents a non-fermented extract and B represents a fermented extract. Most of the mulberry leaf alkaloids were consistent with most reports of 1-deoxynojirimycin (1-DNJ) (Lou et al. , 2011, Kim et al . , 2003, Kojima et al . , 2010, Kimura et al 1995). 1-deoxynojirimycin (1-DNJ) was isolated and its content was measured by preparative TLC (PTLC). As a result, the fermentation product ('Solid-state fermentation' (1-deoxynojirimycin, 1-DNJ) was obtained from mulberry leaves by 2.08 g per 100 g of dried mulberry leaf powder and 8.08-fold higher than that of non-fermented product (non-fermentation in Table 7) It was judged that the most effective method to obtain a large amount of 1-deoxynojirimycin was about 70% in the content of piperidine alkaloids.

Example  5. Alpha Glucosidase (? glucosidase ) Inhibitory activity

Alpha glucosidase inhibitory activity was measured according to the method of Hsieh et al. (2010). Fermentation and non-fermentation 1 ~ 5% of piperidine alkaloid was dissolved in purified water and then filtered under reduced pressure to remove residues. 50 ml of the supernatant was dissolved in 50 ml of 0.15 U / ml α-glucosidase enzyme solution, 200 mmol potassium phosphate (pH 6.8), preincubated at 37 ° C for 10 min, and then added with 100 ml of 3 mmol p- NPG (p-nitrophenyl-glucopyranoside) dissolved in 0.1 molar phosphate buffer (pH 7.0) Lt; / RTI > The reaction was stopped with 750 ml of 0.1 mol Na ₂ CO ₃ and the absorbance was measured at 405 nm. In order to increase the utilization of mulberry leaf, the same inhibitory activity was measured by hot water extraction of fermented mulberry leaf and non - fermented product.

Α-glucosidase is an enzyme present in the brush-border membrane of small intestine epithelium. It acts as a monosaccharide which is easily digested and absorbed by carbohydrates. The inhibitory effect on α-glucosidase is the main target enzyme for the measurement of anti-diabetic activity because it can suppress the increase of blood glucose after the carbohydrate diets. The results of comparing the inhibitory activity of alpha-glucosidase of the pepperidin alkaloid extracted from the non-fermented product and the fermented product with that of the hot-water extract for comparison thereto are shown in FIG. 1% of the piperidine alkaloids extracted from the non-fermented product ('Non-fermented mulberry leaves' of FIG. 4) and the fermentation product ('Solid-state fermented mulberry leaves of A' 70.6% and 73.2%, respectively. The addition of 3%, 90.2%, 93.0% and 5% inhibition showed 95.0% and 97.0% inhibition activity, respectively, but there was no significant difference between nonfermented and fermented products. It was concluded that the composition of piperidine alkaloid contained in mulberry leaves did not show any significant difference between fermentation and non - fermentation.

On the other hand, the α-glucosidase inhibitory activity of the non-fermented product and the fermented product in the hot-water extract was 7.7 (%) when 1% of the non-fermented hot-water extract (non-fermented mulberry leaves in FIG. 4) (B) Solid-state fermented mulberry leaves of fermented hot-water extract (B) were added at 1%, 32.5% and 3%, respectively. 65.7%, and 84.7% when 5% was added. In the case of hydrothermal extraction, the enzyme inhibitory activity showed a large difference due to the difference in the content of piperidine alkaloid between the fermentation and the non-fermented product. The novel Lactobacillus plantarum TO-2100 ( Lactobacillus plantarum TO-2100) fermentation by the fermentation of the mulberry leaves.

The invention of the novel mulberry Such a Lactobacillus Planta room TO-2100 (Lactobacillus plantarum TO -2100 , accession number KFCC 11537P) alpha-glucosidase (α-glycosidase) a carbohydrate metabolizing enzyme obtained by fermentation by using a solid-state A nitrogen-containing compound having an inhibitory activity, and a fermented mulberry leaf having increased contents such as piperidine alkaloids.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the invention. Therefore, it is to be understood that the present invention is not limited to the above-described embodiments. Accordingly, the true scope of the present invention should be determined by the technical idea of the appended claims. It is also to be understood that the invention is to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the invention as defined by the appended claims.

Korea Microorganism Conservation Center (Domestic) KFCC11537P 20120717

Attach an electronic file to a sequence list

Claims (8)

2100 ( Lactobacillus plantarum TO-2100 , accession number: 1) having an ability to produce mulberry leaf fermented product having an increased content of 1-Deoxynojirimycin and Piperidine Alkaloid, KFCC 11537P). The method according to claim 1,
Lactobacillus Planta room TO-2100, characterized in that it contains the fiber-decomposing activity (Lactobacillus plantarum TO -2100, accession number KFCC 11537P). A method of fermenting mulberry leaves, which comprises fermenting mulberry leaves using the Lactobacillus planta TO-2100 ( Lactobacillus plantarum TO-2100 , Accession No. KFCC 11537P) of claim 1. The method of claim 3,
Wherein the fermentation is a solid fermentation. delete The method of claim 3,
Wherein the initial moisture content of the fermentation is 20%, the fermentation temperature is 30 to 35 ° C, and the fermentation relative humidity is 60 to 70%. Characterized in that it is fermented by a fermentation method of mulberry leaf using Lactobacillus plantarum TO-2100 ( Lactobacillus plantarum TO-2100, Accession No. KFCC 11537P) according to any one of claims 3, 4 and 6 Fermented mulberry leaves. A food composition comprising the fermented mulberry leaf of claim 7.

Images