WO2013073722A1 - Novel gluconacetobacter sp. gel_sea623-2 isolated from tangerine juice, and preparation method for cellulose gel using same - Google Patents

Abstract

The present invention relates to a Gluconacetobacter sp. gel_SEA623-2 which is isolated from a tangerine juice and is used for the preparation of a cellulose gel, and a preparation method for a cellulose gel using the same. The cellulose gel prepared by the present invention is soft and contains a large amount of moisture, thereby facilitating formulation into a novel form, and is nontoxic and has similar morphological and structural characteristics as an epithelial cell, thereby enabling application as various forms of materials such as a mask sheet, functional cosmetics and the like.

Description

Novel gel_SEA6233-2 strain of Gluconacetobacter isolated from citrus juice and method for preparing cellulose gel using same

The present invention relates to a novel Gluconacetobacter sp. Gel_SEA623-2 strain isolated from citrus juice and used for the preparation of cellulose gel, and a method for producing cellulose gel using the same.

Cellulose is one of the most abundant biological materials in nature. Especially, cellulose produced by acetic acid bacteria has attracted much attention not only as an additive of food but also as a new value-added industrial new material.

Since it has been reported that acetic acid bacteria produce cellulose (Brown. AJ, J. Chem. Soc., Vol. 49, pp. 432-439, 1986), microbial cellulose (Bacterial Cellulose) is a new material. Has not been studied. Plant-derived cellulose and bacterial cellulose show a great structural difference. That is, the microbial-derived bacterial cellulose is composed of ribbon-like bundles, while the plant-derived cellulose is composed of bundles of microfibrils.

In particular, cellulose produced by acetic acid bacteria is produced in a pure state that does not contain lignin or hemicellulose, unlike plant-derived cellulose composed of bundles of microfibrils. As a result of research on productivity improvement, genetic manipulation, establishment of culture conditions by improving the strain, it can be used as advanced materials such as industrial materials for high strength, composite fibers, medical materials and enzyme immobilization.

Current strains producing cellulose include Acetobacter sp., Agrobacterium sp., Rhizobium sp., Pseudomonas sp., And Sarcina sp. ) are hollow, especially acetonitrile bakteo Here Come been a lot num (Acetobacter xylium), acetonitrile bakteo wave Uriah Stephen Augustine (A. pasteurinanus) and acetonitrile bakteo century you (A. hansenii) in acetonitrile in bakteo (Acetobacter sp.). Recently, 16S rRNA sequencing has been used to reclassify the molecular system, and the strain producing high-purity cellulose is morphologically enterococci and strong acid resistance bacteria, which is reported to secrete cellulose from cells. It is.

The most common carbon source used in the production of cellulose is glucose (Masaoka S, et al., J. Fermen. Bioeng., 75, 18-22, 1993), and other materials such as citrus and persimmon vinegar (Kim SY). , et al., Appl Biochem Biotechnol., 129-132, 705-15, 2006), apple juice (Lee OS, et al., J. Kor. Soc. Food Sci. Nutr., 31 (4), 572-577 , 2002), grape juice (Son CJ, et al., J. Appl. Microbiol. Biotechnol., 12 (5), 722-728, 2002), beer waste (Park JK, et al., Kor Chem Eng Res., 44 (1), 52-57, 2006) and coconut byproducts (Jonas R, et al., Polym Degrad Stab., 59, 101-106, 1998).

Citrus fruits, on the other hand, are a fruit that contains a lot of multi-functional medicinal ingredients, and have been used as herbal medicines for a long time in Korea. At the same time, it has been used as a health food material for nutritional sources such as vitamins, dietary fiber, organic acids and free sugars. In addition, citrus fruits contain various biologically active substances such as flavonoids, carotenoids, coumarins, phenylpropanoids, limonoids, pectin, celluloses, hemicelluloses, and the like. About 60 flavonoids are known and most exist in glycosides associated with sugars. The flavonoids are known for homeostasis, prevention of circulatory diseases, anti-inflammatory, anti-allergic, antibacterial, antiviral, hypolipidemic, immune enhancing action.

Although Japanese Patent Nos. 3,341,017 and 4,061,661, Korean Patent No. 405,776 and the like have disclosed a method for producing cellulose using microorganisms of the genus Acetobacter , a film having a thickness that is commercially available from fruit juices such as citrus fruits. A method for preparing the cellulose gel in form has not been developed yet.

Thus, the present inventors conducted a study on the various uses of citrus fruits to isolate the new strain from the tangerine, using the strain from a specific medium to confirm that the biocompatibility and physical properties of the cellulose gel can be produced by using the strain To complete.

Accordingly, it is an object of the present invention to provide a novel strain characterized in that it is used to prepare a cellulose gel.

Another object of the present invention is to provide a method for preparing a cellulose gel having excellent biocompatibility and physical properties by inoculating the novel strain in a juice medium.

The present invention is characterized by the novel Gluconacetobacter sp. Gel_SEA623-2 strain, which is used to prepare cellulose gels.

In another aspect, the present invention is characterized by a method for producing a cellulose gel by inoculating the strain in the juice medium.

In another aspect, the present invention is characterized by a cellulose gel that can be manufactured by the above production method can be applied for various uses.

Cellulose gel prepared according to the present invention is soft in strength and contains a large amount of moisture, so it is easy to formulate in a new form, there is no toxicity, and similar to epithelial cells in morphological and structural features, mask packs, functional cosmetics It can be used in various forms of materials.

Figure 1 shows the Gluconacetobacter sp. Gel_SEA623-2 strain isolated from the natural fermentation of citrus juice.

Figure 2 is a scanning electron micrograph (SEM) of the gel_SEA623-2 strain of gluconacetobacter genus.

Figure 3 is a photograph of the citrus cellulose gel prepared by Example 2.

Figure 4 is a SEM photograph of the citrus cellulose gel prepared by Example 2.

5 is a photograph confirming whether the reference strain can form a citrus cellulose gel.

6 is a SEM photograph of a coconut cellulose gel that is commercially available.

Figure 7 is a photograph confirming the production of cellulose gel for each juice medium.

8 is a graph comparing the thickness of the prepared cellulose gel for each juice medium.

Figure 9 is a photograph comparing the citrus cellulose gel prepared by varying the pH of the citrus juice.

Figure 10 is a graph comparing the thickness of the citrus cellulose gel prepared by varying the pH of the citrus juice.

Figure 11 is a photograph comparing the citrus cellulose gel prepared by varying the sugar content of the citrus juice.

12 is a graph comparing the thicknesses of citrus cellulose gels prepared by varying the sugar content of citrus juices.

Figure 13 is a photograph comparing the citrus cellulose gel prepared by varying the incubation temperature.

14 is a graph comparing the thickness of the citrus cellulose gel prepared by varying the incubation temperature.

15 is a photograph confirming the citrus cellulose gel prepared by using the citrus fruit fermentation broth as a medium.

Figure 16 is a graph comparing the thickness of the citrus cellulose gel prepared by varying the glycolysis enzyme of the citrus fruit fermentation broth.

Referring to the present invention in more detail as follows.

Gluconacetobacter sp. Gel_SEA623-2 strain, which is used to prepare the cellulose gel of the present invention, is a new strain isolated from citrus juice, and it is dated February 4, 2010, at the Microbial Bank of Agricultural Genetic Resource Information Center. In addition, the new strain Gluconacetobacter sp. Gel_SEA623-2 has been deposited with the Korea Biotechnology Research Center (KCTC), an international depository institution, on July 4, 2011. Deposited (Accession Number: KCTC 11977BP).

In another aspect, the present invention includes a method for producing a cellulose gel comprising the step of inoculating the strain Gluconacetobacter sp. Gel_SEA623-2 in the juice medium. It is preferable to inoculate the gel_SEA623-2 strain of the gluconacetobacter in the juice medium and incubate while maintaining the temperature at 20 ~ 30 ℃ in aerobic conditions of aseptic acid, and culture for 7 to 21 days.

The juice medium is not particularly limited as long as it is a liquid juice that can be obtained from a general fruit, preferably citrus fruits, pears, apples, grapefruit, etc. may be used, and more preferably citrus fruits are used.

In the present specification, "citrus fruits" refers to citron grown in the south of Europe, as well as citrus grown in the south of Europe, lemon (lemon) grown in the US and the like, paragraphs or grapefruits grown in the tropics, etc. Cultivars grown in the world, such as grape fruit, sour oranges grown in India, Italy, India, Italy, etc. It is meant to include all kumquat or citron (한 子), Hallabong, jinji flavor, tangerine, wild mandarin, blue mandarin orange, tangerine, sugar citrus, red mandarin orange, tangerine, soybean persimmon, persimmon, tangerine. Preferably, it means Hallabong, kumquat and true flavor that are easy to ferment by the yeast because of the high wet or sweet sugar used in the following examples.

The strain is preferably inoculated in the juice medium by pre-cultivation at 25 ~ 35 ℃, pH 2.5 ~ 5.0 conditions, more preferably 26 ~ 30 ℃, pH 3.0 ~ 4.0 range.

The juice medium preferably has an acidity in the pH range of 2.5 to 5.0, more preferably in the range of pH 3.0 to 4.0. In addition, the juice medium is preferably used in the range of 1 to 30 brix sugar, more preferably in the range of 5 to 20 brix.

In addition, as the juice medium, not only citrus juice, but also a fermentation broth obtained by treating and fermenting a glycolysis enzyme on the gourd produced after the juice of the citrus can be used, the fermentation broth also preferably has an acidity in the pH range of 2.5 to 5.0, More preferably, those in the range of pH 3.0 to 4.0 are used. In addition, the fermentation broth uses a sugar in the range of 1 to 30 brix, preferably in the range of 5 to 20 brix. The glycolysis can be used enzymes well known in the art, commercially known Pectinex 100L, Viscozyme L, AMG 300L, Celluclast 1.5L and the like can be used. Table 1 shows the characteristics of the commercial synthase.

	Pectinex 100L	Viscozyme L	AMG 300L	Celluclast 1.5L
origin	Aspergillus Niger ( Aspergillus niger )	Aspergillus Aculitus ( Aspergillus aculeatus )	Aspergillus Niger ( Aspergillus niger )	Tricoderma Reise ( Trichoderma reesei )
Specialization	pectolytic enzyme (pectintranseliminase, polygalacturonase, pectinesterase)	multi-enzyme complex (arabanase, cellulase, β-glucanase, hemicellulase, xylanase)	1,4-α-D-glucosidase (glucoamylase)	cellulase (the breakdown of cellulose into glucose, cellobiose and higher glucose polymers)

Citrus gourd is preferably fermented for about 3 to 12 hours after inoculation of the glycolytic enzyme, and it is preferable to ferment each of the enzymes to an optimal temperature, and to maintain about 40 to 70 ° C.

Cellulose gel prepared by the manufacturing method shows a commercially available thickness range of 5 ~ 20 mm, and has a tensile strength of 5 ~ 30 N / cm ^{2 It} is possible to obtain a relatively soft gel, 70 ~ 95% by weight High water content. In particular, the cellulose gel is excellent in antibacterial and economically very advantageous using a natural material, there is no toxicity at all, it can be usefully used as a functional cosmetics.

The cellulose gel can be used as a component of the cosmetic powder itself or by powdering it. The cellulose gel-containing cosmetic may be prepared in liquid or solid form using bases, adjuvants and additives commonly used in the cosmetic field. Cosmetics in liquid or solid form may include, but are not limited to, for example, cosmetics, creams, lotions, baths and the like. Bases, auxiliaries and additives commonly used in the cosmetic field are not particularly limited and may include, for example, water, alcohols, propylene glycol, stearic acid, glycerol, cetyl alcohol and liquid paraffin.

Hereinafter, the present invention will be described in detail based on the following examples, but the present invention is not limited thereto.

Example 1 Selection of Gel Production Strains from Citrus Fruits

In order to isolate the gel-producing strains, citrus juice was used to culture at 15 ° C. for 15 days, followed by spontaneous fermentation for 14 days. At this time, the composition of the Glunobacter oxydans plate medium was 10% glucose, 1% yeast extract, 2% CaCO ₃ , 1.5% agar, 1% EtOH (pH 6.8).

The entire nucleotide sequence (958 bp, SEQ ID NO: 1) of the isolated single strain 16S rRNA was determined (Juke TH, Cantor CR: Evolution of protein molecules.In mammalian protein metabolism, Edited by HN, Munro p21, Academic Press. New York, 1969) As a result of analyzing the percent similarity and phylogenetic tree, as shown in Table 2 it showed more than 99% homology with the strains of the genus Gluconacetobacter. Therefore, the strain was named Gluconacetobacter sp. Gel_SEA623-2, and was deposited on February 4, 2010 in the Microbial Bank of Agricultural Genetic Resources Information Center (Accession Number: KACC 91526P). Gluconacetobacter sp. Gel_SEA623-2 strain was deposited on July 4, 2011 to the Korea Institute of Bioscience and Biotechnology (KCTC), an international depository institution (accession number: KCTC 11977BP).

Standard strain	Similarity
Ochrobactrum sp. BB-1	99% (956/958)
Gluconacetobacter kombuchae	99% (956/958)
Gluconacetobacter maltiaceti	99% (951/958)
Gluconacetobacter persimmonis	99% (951/958)
Gluconacetobacter entanii	99% (951/958)

On the other hand, Figure 1 is a photograph of the Gluconacetobacter sp. Gel_SEA623-2 strain isolated from the natural fermentation of citrus juice. In addition, Figure 2 is a photograph of the strain observed with a scanning electron microscope, the strain was confirmed to be 1.5 ~ 3 ㎛ size bacilli.

Experimental Example 1: (. Gluconacetobacter sp) gluconate acetonitrile bakteo in biochemical properties of the strain gel_SEA623-2

To compare the biochemical properties of isolated strains, Gluconacetobacter liquefaciens KACC 12360 (= ATCC 14835, IFO 12388), Gluconacetobacter hansenii KACC 12359 (= ATCC 35959, IFO 14820), Gluconacetobacter diazotrophicus KACC 12358 (= ATCC 49037), Gluconobacter oxydans KACC 11292 (= ATCC 19357, IFO 14819) and Acetobacter trophic ( Actobacter trophus) ) KACC 12236 (= IFO 16470) was purchased from the Korea Agricultural Microbial Resources Center (KACC), Gluconacetobacter xylinus subsp. Xylinus KCCM 40198 (= ATCC 23768, IFO 14814), Gluconacetobacter xylinus subsp. Xylinus KCCM 40231 (= ATCC 23767, IFO 15237), Gluconacetobacter xylinus subsp. Xylinus KCCM 40216 (= ATCC 10245, IFO 3173) and Acetobacter xylinum KCCM 40407 (= IFO 3288) were purchased from the Korean Breeders Association (KCCM) and compared with these reference strains. The results of comparing the biochemical properties of are shown in Table 3 below.

	KCTC 11977BP 1)	KACC 12360 2)	KACC 12359 3)	KACC 12358 4)	KACC 11292 5)	KACC 12236 6)	KCCM 40198 7)	KCCM 40231 8)	KCCM 40216 9)	KCCM 40407 10)
ONPG (β-galactosidase)	-	-	-	-	-	-	-	-	-	-
Arginine Dehydrolase	-	-	-	-	-	-	-	+	-	-
Lysine Decarboxylase	-	-	-	-	-	-	-	-	-	-
Ornithine Decarboxylase	-	-	-	-	-	-	-	-	-	-
Citric acid use	-	-	-	-	-	-	-	-	-	-
H2S production	-	-	-	-	-	-	-	-	-	-
Uraase	-	-	-	-	-	-	-	-	-	-
Tryptophan Deaminase	-	-	-	-	-	-	-	-	-	-
Indole production	-	-	-	-	-	-	-	+	-	-
VP test	-	-	+	+	-	+	+	-	+	+
Gelatin liquefaction	-	-	-	-	-	-	-	-	-	-
Production of Acids from Glucose	+	+	+	+	+	+	+	+	+	+
Generation of acid from mannitol	-	-	-	-	-	-	-	-	-	-
Generation of Acids from Inositol	-	-	-	-	-	-	-	-	+	-
Generation of Acids from Sorbitol	-	-	-	+	-	-	-	-	-	-
Production of Acids from Rhamnose	+	-	-	+	-	-	+	-	+	+
Production of acids from saccharose	-	+	-	+	+	-	+	+	+	+
Production of Acids from Melibiose	+	+	+	+	+	+	+	+	+	+
Production of Acids from Amigdaline	-	-	-	-	-	-	-	-	+	-
Production of acids from arabinose	+	+	+	+	+	+	+	-	+	+
NO2 production	-	-	-	-	-	+	-	-	-	-
N2 gas reduction	-	-	-	-	-	-	-	-	-	-
cellulose production	+	-	-	-	-	-	+	-	-	-
-, voice; +, Positive 1) Gluconacetobacter sp. gel_SEA623-2 2) Gluconacetobacter liquefaciens 3) Gluconacetobacter hansenii 4) Gluconacetobacter diazotrophicus 5) Gluconobacter oxydans 6) Acetobacter trophicus 7) Gluconacetobacter xylinus subsp. xylinus 8) Gluconacetobacter xylinus subsp. xylinus 9) Gluconacetobacter xylinus subsp. xylinus 10) Acetobacter xylinum

Gluconacetobacter genus gel_SEA623-2 strain isolated in Example 1 showed a positive reaction only in the fermentation using glucose, rhamnose, melibiose and arabinose, among the strains of the genus Gluconacetobacter and acetobacter Strains showing the same biochemical properties were not identified.

Example 2 Preparation of Cellulose Gel from Citrus Juices

Inoculated with SEA623-2 strain of Gluconacetobacter isolated in Example 1 in a sugar juice of 10 brix (10% (v / v)) of pH 10 brix, pH 3.0, 1 × 10 ⁵ cell / ml, Incubated at 25 ° C. for 20 days under aerobic conditions. As a result, the initial gel formation rate of the strain was very good, and the gel membrane began to form from the second day of culture, and the photo of the citrus cellulose gel produced after 20 days was shown in FIG. 4 is shown.

Experimental Example 2 Preparation of Cellulose Gel Using a Reference Strain

The gel was induced under the same conditions as in Example 2 with respect to the reference strain compared in Experimental Example 1, but out of a total of nine reference strains, only Gluconacetobacter xylus subsp. Only Xylus KCCM 40198 strain formed cellulose gel, and the remaining strains did not form cellulose gel (FIG. 5).

However, the gluconacetobacter xylinus subsp. In the case of the Xylus KCCM 40198 strain, the gel membrane began to form only after 3 days of culture, and after 20 days, the thickness and physical properties of the gel were compared with those of the cellulose gel of Example 2, and are shown in Table 4 below. Tensile strength of cellulose gel was measured at 25 ° C. using XT-RA Stable Micro Systems according to the KS standard, and the prepared citrus cellulose gel was completely dried in an oven at 50 ° C. and then compared before and after drying. Moisture content was measured.

	Gluconacetobacter sp. gel_SEA623-2 (KCTC 11977BP)	Gluconacetobacter xylinus subsp. xylinus (KCCM 40198)
Thickness (mm)	8.0	2.5
Tensile Strength (N / cm 2 )	11.23 ± 0.35	18.78 ± 5.45
Moisture content (%)	90.20	88.35

As shown in Table 4, the glucoacetobacter xylinus subsp. In the case of Xylus KCCM 40198 strain, the thickness thereof was very thin, and the tensile strength of the gel was high.

Comparative Example: Preparation of Cellulose Gel from Coconut

Gluconacetobacter xylinus subsp., Widely known as a strain capable of producing cellulose gels. Xylus KCCM 40198 was inoculated at 1 × 10 ⁵ cell / ml in a medium composition (10% (v / v)) consisting of 89.5% by weight of coconut juice, 10% by weight of sugar and 0.5% by weight of vinegar, and at 25 ° C. for 20 days. Coconut cellulose gel was prepared by culturing in aerobic conditions, and the photograph of observing the prepared cellulose gel with a scanning electron microscope is shown in FIG. 6.

As shown in FIG. 6, the citrus cellulose gel of the present invention has a thin fiber and irregularly bundles of fibers, so that the space between the fibers is 250 to 500 compared to the coconut cellulose gel (100 to 250 nm) of FIG. 6. It was confirmed that the appearance was large as nm. Based on this, the cellulose gel of the present invention was relatively low in density, and thus easy to formulate. Therefore, the cellulose gel of the present invention was confirmed to exhibit characteristics that are easy to use as a cosmetic material such as a mask pack, lotion, and cream.

In addition, the coconut cellulose gel is attached to the fiber bundles in a large amount of components that appear to be agglomerate, but in the citrus cellulose gel, these components were hardly shown, and thus it was confirmed that a more pure cellulose gel could be prepared.

Example 3 Preparation of Cellulose Gel from Juice Medium

On the other hand, orange juice (10% (v / v)) of 10 brix, pH pH 3.2, grapefruit juice (10% (v / v)) of 10 brix, pH 2.7, sugar 10 brix, pH pH Inoculated with the gluconacetobacter genus gel_SEA623-2 strain in the juice of 3.0 pear juice, sugar 10 brix, acidity pH 3.0 to prepare the gel (FIG. 7), the thickness after 20 days It is shown in Table 5 and FIG.

	Juicer badge
	Wenzhou Citrus	Orange	grapefruit	stomach	Apple
Thickness (mm)	8.0	7.7	5.9	6.1	1.0

As shown in Table 5 and Figure 8 it was able to prepare a cellulose gel of a certain thickness from a number of juice medium, in particular, it can be confirmed that the thickest cellulose gel can be prepared from the juice of Wenju juice.

Example 4: Confirmation of gel formation according to pH of citrus juice

The thickness of the gel produced by preparing the gel in the same manner as in Example 2 except for changing the pH of the juice of the hot-swallowed citrus juice with NaOH, acetic acid to pH 2.0, 2.5, 3.0, 3.5, 4.0, 5.0 It measured, and the result is shown in Table 6 and FIGS.

	PH of Wenju juice
	2.0	2.5	3.0	3.5	4.0	5.0
Thickness (mm)	0.0	6.1	8.0	7.4	7.0	5.2

As shown in Table 6, it was confirmed that the cellulose gel is produced thickest in the vicinity of pH 3.0 ~ 3.5.

Example 5: Confirmation of gel formation according to the sugar content of citrus juice

The thickness of the gel produced by measuring the gel was produced in the same manner as in Example 2 except that the sugar content of the juice of Wenju citrus was adjusted to 1, 5, 10, 20, 30 brix. The results are shown in Table 7 and FIGS. 11 to 12.

	Citrus juice medium sugar
	One	5	10	20	30
Thickness (mm)	2.1	5.4	8.0	5.2	5.4

As shown in Table 7, it was confirmed that the cellulose gel can be produced well in the range of 1 to 30 brix.

Example 6: Confirmation of gel production according to the culture temperature

Except for varying the temperature at 20, 30, 35, 40 ℃ in the culture was prepared in the same manner as in Example 2 to measure the thickness of the gel produced, the results are shown in Table 8 and 13 to 14 Indicated.

	Incubation temperature (℃)
	20	30	35	40
Thickness (mm)	8.3	9.3	7.7	0.0

As shown in Table 8, it was confirmed that the cellulose gel is produced thickest in the vicinity of 20 ~ 30 ℃.

Example 7 Preparation of Citrus Cellulose Gel from Citrus Foil

100 g of commercial Pectinex 100L and Viscozyme L enzymes were inoculated into 50 kg of the produced byproducts after the juice of Wenju citrus persimmon and fermented at 50 ° C. for 10 hours to obtain a fermentation broth. As a result of measuring the sugar and acidity of the fermentation broth it was confirmed that the 12 brix, pH 3.5.

Gluconacetobacter genus gel_SEA623-2 strain isolated in Example 1 was inoculated at 10% (v / v) of the fermentation broth at 1 × 10 ⁵ cell / ml, and cultured at 25 ° C. for 20 days under aerobic conditions. A photo of the citrus cellulose gel produced after 20 days has been shown in FIG. 15, and the thickness of the gel over time is shown in FIG. 16.

As shown in FIG. 15, a cellulose gel can be prepared from the fermentation broth of Wenju's persimmon foil using gel_SEA623-2 strain, and the thickness of the gel reaches about 8.0 mm, which is significantly different from that of the juice solution itself. Not me.

Experimental Example 3 Measurement of Physical Properties of Cellulose Gel

The tensile strength of the citrus cellulose gels prepared in Examples 2 and 5 was measured at 25 ° C. using XT-RA Stable Micro Systems in accordance with the KS standard. In addition, the prepared citrus cellulose gel was completely dried in an oven at 50 ° C., and the moisture content was measured by comparing the mass before and after drying. The physical property measurement results of the gel prepared in each example are shown in Tables 9 to 12 below.

	Juicer badge
	Wenzhou Citrus	Orange	grapefruit	stomach	Apple
The tensile strength (N / cm 2 )	11.23 ± 0.35	4.87 ± 1.45	3.09 ± 0.90	2.76 ± 0.45	-
Moisture content (%)	90.20	88.02	83.23	89.43	-

	PH of Wenju juice
	2.0	2.5	3.0	3.5	4.0	5.0
The tensile strength (N / cm 2 )	-	6.98 ± 0.39	11.23 ± 0.35	15.25 ± 0.34	17.51 ± 0.34	8.94 ± 0.22
Water content (%)	-	90.60	90.20	88.20	87.62	93.06

	Citrus juice medium sugar
	5	10	20	30
The tensile strength (N / cm 2 )	17.46 ± 0.44	11.23 ± 0.35	7.78 ± 0.62	3.82 ± 0.60
Moisture content (%)	90.23	90.20	86.42	85.46

	Incubation temperature (℃)
	20	30	35	40
The tensile strength (N / cm 2 )	29.48 ± 0.41	13.06 ± 0.70	4.687 ± 0.59	-
Moisture content (%)	93.58	90.32	90.52	-

As shown in Tables 9 to 12, it can be seen that the cellulose gel of the present invention has a high moisture content of 80% or more, and exhibits a soft property by showing a low tensile strength as compared to commercially available bio cellulose gel. .

On the other hand, in the case of the cellulose gel prepared from the coconut cellulose gel prepared by the comparative example and the citrus foil according to Example 6 was also measured in the same manner as above the tensile strength and water content, the results are shown in Table 13 below .

	Comparative example	Citrus Foil Cellulose Gel
		pectinex Treated Fermentation Broth	viscozyme Treated Fermentation Broth
The tensile strength (N / cm 2 )	63.87 ± 0.26	8.95 ± 1.26	11.89 ± 1.11
Water content (%)	88.54	90.69	90.15

As shown in Table 13, even when the gel was prepared using the citrus fruit fermentation broth as a medium, it could be confirmed that a cellulose gel having similar physical properties as in the case of using the citrus juice could be prepared.

Experimental Example 4: grinding properties according to the tensile strength of the cellulose gel

In order to check the physical properties of the cellulose gel prepared in Example 2 and the cellulose gel having a tensile strength of more than 20 N / cm ² after grinding, the prepared cellulose gel was ground for 30 seconds using a blender of 10,000 rpm. The results are shown in FIG.

As shown in FIG. 17, the cellulose gel of Example 2 was adjusted to a particle size of 45 μm or less, but when the tensile strength exceeded 20 N / cm ² , such particle size control was impossible. Therefore, the citrus cellulose gel of the present invention can be used as a cream-like perfume material because it can be crushed into particles of a micrometer unit, it can be applied to various formulations.

[Accession number]

Depositary Name: Korea Institute of Bioscience and Biotechnology Biological Resource Center

Accession number: KCTC 11977BP

Deposit date: 20110704

[Revisions under Rule 91 20.01.2012]

<110> REPUBLIC OF KOREA (MANAGEMENT: RURAL DEVELOPMENT ADMINISTRATION)

<120> A novel strain of Gluconacetobacter sp. gel_SEA623-2 from citrus

         juice and cellulose gel by using it

<160> 1

<170> KopatentIn 1.71

<210> 1

<211> 958

<212> DNA

<213> Gluconacetobacter sp. gel_SEA623-2

<400> 1

tgggtggggg ataactttgg gaaactgaag ctaataccgc atgacacctg agggtcaaag 60

gcgcgagtcg cctgtggagg aacctgcgtt cgattagcta gttggtgggg taaaggccta 120

ccaaggcgat gatcgatagc tggtctgaga ggatgatcag ccacactggg actgagacac 180

ggcccagact cctacgggag gcagcagtgg ggaatattgg acaatgggcg caagcctgat 240

ccagcaatgc cgcgtgtgtg aagaaggttt tcggattgta aagcactttc agcggggacg 300

atgatgacgg tacccgcaga agaagccccg gctaacttcg tgccagcagc cgcggtaata 360

cgaagggggc aagcgttgct cggaatgact gggcgtaaag ggcgcgtagg cggttgttac 420

agtcagatgt gaaattcccg ggcttaacct gggggctgca tttgatacgt gacgactaga 480

gtgtgagaga gggttgtgga attcccagtg tagaggtgaa attcgtagat attgggaaga 540

acaccggtgg cgaaggcggc aacctggctc atgactgacg ctgaggcgcg aaagcgtggg 600

gagcaaacag gattagatac cctggtagtc cacgctgtaa acgatgtgtg ctggatgttg 660

gatggcttgg ccattcagtg tcgtagttaa cgcgataagc acaccgcctg gggagtacgg 720

ccgcaaggtt gaaactcaaa ggaattgacg ggggcccgca caagcggtgg agcatgtggt 780

ttaattcgaa gcaacgcgca gaaccttacc aggacttgac atgcggaggc tgtgtccaga 840

gatgggcatt tctcgcaaga gacctccagc acaggtgctg catggctgtc gtcagctcgt 900

gtcgtgagat gttgggttaa gtcccgcaac gagcgcaacc ctcgccttta gttgccag 958

Claims (17)

1. Novel Gluconacetobacter sp. Gel_SEA623-2 strain (KCTC 11977BP), characterized in that it is used to prepare cellulose gel from fruit juice.
   
   
2. The method of claim 1, wherein the strain is Gluconacetobacter genus gel_SEA623-2 strain (KCTC 11977BP), characterized in that isolated from citrus.
3. The gluconacetobacter genus gel_SEA623-2 strain (KCTC 11977BP) according to claim 1, wherein the juice is a citrus juice or a citrus fruit fermentation broth.
   
   
4. A method for producing a cellulose gel, comprising inoculating the strain of claim 1 in a juice medium to prepare a cellulose gel.
   
   
5. The method of claim 4, wherein the juice medium is a method of producing a cellulose gel, characterized in that in the range of pH 2.5 ~ 5.0.
   
   
6. The method of claim 5, wherein the juice medium is a method for producing a cellulose gel, characterized in that in the range of pH 3.0 ~ 4.0.
   
   
7. 5. The method of claim 4, wherein the juice medium has a sugar content in the range of 1 to 30 brix.
   
   
   
   
8. 5. The method of claim 4, wherein the juice medium is a citrus juice.
   
   
9. 5. The method of claim 4, wherein the juice medium is a fermentation broth obtained by treating and fermenting a glycolytic enzyme on a gourd produced after the tangerine juice.
   
   
10. The method of claim 4, wherein the strain is a method for producing a cellulose gel, characterized in that the pre-cultured at 25 ~ 35 ℃, pH 2.5 ~ 5.0 conditions.
    
    
11. The method of claim 4, wherein the strain is inoculated into the medium and cultured for 7 to 21 days at 20 ~ 30 ℃.
    
    
12. 10. The method of claim 8 or 9, wherein the tangerine is a method of producing a cellulose gel, characterized in that selected from Wenzhou, persimmon, kumquat and true flavor.
    
    
13. Cellulose gel prepared by the method of any one of claims 4 to 11.
    
    
14. The cellulose gel of claim 13, wherein the cellulose gel has a thickness in the range of 5-20 mm.
    
    
15. The cellulose gel of claim 13, wherein the cellulose gel has a tensile strength in the range of 5 to 30 N / cm ² .
    
    
16. The cellulose gel of claim 13, wherein the cellulose gel contains 70 to 95 weight percent moisture.
    
    
17. The cellulose gel of claim 13, wherein the cellulose gel is made of a cosmetic cream.

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