KR102612094B1 - Novel strain Cryptocline arctostaphyli RN2 having xylan degrading activity - Google Patents

Abstract

The present invention relates to a new strain Cryptocline arctostaphyli RN2 with xylan decomposition activity, a method for decomposing polysaccharides using the same, a composition for decomposing polysaccharides, a method for producing disaccharides, and a composition for producing disaccharides.
According to the present invention, various polysaccharides, including xylan, can be decomposed very effectively through biological methods, and in particular, disaccharides can be effectively produced from xylan.

Description

Novel strain Cryptocline arctostaphyli RN2 having xylan degrading activity}

The present invention relates to a new strain Cryptocline Arctostaphylli RN2 having xylan decomposition activity and a method for decomposing polysaccharides using the same, a composition for decomposing polysaccharides, a method for producing disaccharides, and a composition for producing disaccharides.

Xylan is hemicellulose, one of the main components that accounts for about 20 to 40% of the cell walls of land plants, and is composed of arabinosyl, acetyl, and glucuronosyl. It has a complex structure composed of substituted β-1,4-linked xylose units, and can be hydrolyzed by various xylanases.

Xylanase is required in various fields such as the pulp-pre-bleaching process to remove hemicellulose from the pulp, promoting digestion of livestock feed, food and baking additives, fruit and vegetable processing, ethanol and xylitol production, and papermaking. Research on the characteristics of xylanase suitable for the field is required. In addition, xylooligosaccharide (XOS), a xylan hydrolysis product produced by xylanase, is known to have various physiological activities such as antibacterial, antioxidant, and anti-inflammatory, and is used in various fields such as food-pharmaceutical industry, agricultural-livestock industry, and cosmetics industry. It is expected that it can be used.

The decomposition method using a hydrolytic enzyme such as xylanase mentioned above has the advantage of being able to selectively and specifically hydrolyze polysaccharides to produce the target oligosaccharide. In particular, hydrolysis of plant biomass using microbial enzymes is accepted as an environmentally friendly alternative to chemical decomposition methods that produce harmful by-products. For this reason, many researchers are attempting to isolate new microorganisms that produce more suitable enzymes.

Accordingly, the present inventors sought to discover new microorganisms that could be used to biologically decompose (enzymatically) decompose xylan and produce specific decomposition products from xylan.

Christakopoulos P, Katapodis P, Kalogeris E, Kekos D, Macris BJ, Stamatis H, Skaltsa H. Antimicrobial activity of acidic xylo-oligosaccharides produced by families 10 and 11 endoxylanases. Int J Biol Macromol. 2003 Jan 15;31(4-5):171-5.

Therefore, the main purpose of the present invention is to provide a new microorganism that can be used to biologically decompose xylan and produce specific decomposition products from xylan.

Another object of the present invention is to provide a method of utilizing the microorganism.

Another object of the present invention is to provide a product utilizing the above microorganisms.

According to one aspect of the present invention, the present invention provides Cryptocline arctostaphyli RN2 strain deposited under accession number KACC 93360P.

According to another aspect of the present invention, the present invention provides a method for decomposing polysaccharides comprising contacting a culture of the strain or a purified product of the culture with a polysaccharide.

In the polysaccharide decomposition method of the present invention, the polysaccharide is preferably xylan, arabinoxylan, or amylose.

According to another aspect of the present invention, the present invention provides a composition for decomposing polysaccharides comprising a culture of the above strain or a purified product of the above culture.

In the composition for decomposing polysaccharides of the present invention, the polysaccharide is preferably xylan, arabinoxylan, or amylose.

According to another aspect of the present invention, the present invention provides a method for producing a disaccharide comprising contacting a culture of the strain or a purified product of the culture with xylan.

According to another aspect of the present invention, the present invention provides a composition for producing a disaccharide comprising a culture of the above strain or a purified product of the above culture.

According to the present invention, various polysaccharides, including xylan, can be decomposed very effectively through biological methods, and in particular, disaccharides can be effectively produced from xylan.

Figure 1 shows the NJ (Neighbor-Joining) phylogenetic tree analyzed based on the ITS-D1D2 sequence of the strain of the present invention.
Figure 2 shows a photograph taken through a microscope of the bacterial cells of the strain of the present invention grown on a solid medium (left) and a photograph taken of colonies of the strain of the present invention formed on a solid medium (right). In the colony photo on the right, the left side (marked '(top)') is a shot of the top of the medium, and the right side (marked '(bottom)') is a shot of the bottom of the medium. .
Figure 3 shows the results of a polysaccharide decomposition activity test of the strain of the present invention. (A), amylose decomposition activity test results; (B), xylan decomposition activity test results; (C), Arabinoxylan decomposition activity test results.
Figure 4 shows the results of xylanase activity experiments according to culture time of the strain of the present invention.
Figure 5 shows the results of TLC analysis of the reaction product of the crude enzyme solution and xylan of the strain of the present invention. G, glucose; X2, xylobiose; X4, xylotetraose; X6, xylohexaose; R, reactant.
Figure 6 shows the microbial deposit of the strain of the present invention.

The new strain Cryptocline arctostaphyli RN2 of the present invention was isolated from rose flowers and deposited in the Korean Agricultural Culture Collection (KACC) of the National Institute of Agricultural Sciences under the accession number KACC 93360P.

The strain of the present invention has the ITS sequence of SEQ ID NO: 1 and the D1D2 sequence of SEQ ID NO: 2. The ITS sequence and D1D2 sequence are representative sequences used to identify fungi, and the ITS sequence and D1D2 sequence of the strain of the present invention are different compared to the ITS sequence and D1D2 sequence of the previously known cryptocline Arctostaphylli.

The strain of the present invention can decompose polysaccharides such as xylan, arabinoxylan, and amylose, and in particular, can decompose xylan to produce disaccharide, that is, a dimer (DP2) consisting of two monosaccharide molecules, as the main product.

The cryptocline Arctostaphylli was isolated as an endophyte of Arctostaphylos uva-ursi in Switzerland in 1984 and its existence was known, but until recently, there was no research using this strain worldwide. In addition, it is a domestically unrecorded species for which no strain isolation or research results have been published in Korea.

The strain of the present invention can exhibit the above-mentioned polysaccharide decomposition activity and the activity of producing disaccharide from xylan outside the cell. This was confirmed through the results of experiments on the decomposition activity of polysaccharides (including xylan) using bacterial removal culture media, suggesting that enzymes related to this activity, such as xylanase, can be secreted to the outside of the cell.

The strain of the present invention can be cultured according to a conventional culture method for culturing fungi, especially fungi of the Cryptocline genus, especially fungi of the Cryptocline arctostaphyli species. For example, for solid culture, PDA (potato dextrose agar) medium can be used, and for liquid culture, PDB (potato dextrose broth) medium can be used, and the culture can be done under temperature conditions of 20 to 40°C.

Based on the characteristics of the strain of the present invention as described above, the present invention provides a method of decomposing polysaccharides comprising the step of contacting a culture of the strain of the present invention or a purified product of the culture with a polysaccharide.

In the present invention, a culture of the present strain can be obtained by culturing the present strain according to the culture method described above. Preferably, the culture of the strain of the present invention is obtained by culturing in PDB medium. More preferably, the culture of the strain of the present invention is obtained by culturing in PDB medium at 28 to 32 ℃ for 2 to 4 days, and more preferably, it is obtained by culturing at 29 to 31 ℃ for 60 to 84 hours. will be. According to this, a culture with higher polysaccharide decomposition activity can be obtained. The culture of the strain of the present invention is preferably obtained by removing the bacterial cells from a culture grown in a liquid medium. This can be achieved using conventional methods for removing bacterial cells from the culture, such as centrifugation and filtration.

In the present invention, the purified product of the culture is obtained through the purification process of the culture of the strain of the present invention, and is obtained from a culture of microorganisms, especially fungi, especially fungi of the Cryptocline genus, especially fungi of the Cryptocline Arctostaphyll species, to the outside of the cell. It may be obtained through a conventional purification process used to purify secreted proteins, especially enzymes, especially glycolytic enzymes. For example, it may be obtained by filtering the culture medium from which the bacterial cells have been removed with a 10KDa cut-off filter.

In the polysaccharide decomposition method of the present invention, the step of contacting the polysaccharide is preferably a step of contacting the culture of the strain of the present invention or a purified product of the culture with xylan, arabinoxylan, or amylose, more preferably contacting the culture with xylan. It's a step.

Contact between the culture of the strain of the present invention or a purified product of the culture and the polysaccharide can be carried out by adding polysaccharide to the culture or purified product, or can be done in a buffer solution. For example, it can also be performed by adding the culture or purification and polysaccharide to a buffer solution. The contact may be carried out, for example, at a temperature of 30 to 40°C, and is preferably carried out at 35 to 39°C.

Based on the above characteristics of the strain of the present invention, the present invention also provides a composition for decomposing polysaccharides comprising a culture of the strain of the present invention or a purified product of the culture.

Details regarding culture and purification in the composition for decomposing polysaccharides of the present invention can be referred to the information described above.

The composition for decomposing polysaccharides of the present invention may further include other ingredients in addition to the culture or purification. For example, ingredients that do not substantially inhibit the decomposition activity of polysaccharides by the culture or purification may further include buffers, diluents, excipients, etc. for the stable action of glycolytic enzymes.

The composition for decomposing polysaccharides of the present invention may include, for example, 0.001 to 100% of the culture or purified product based on the total weight of the composition.

Based on the above characteristics of the strain of the present invention, the present invention also provides a method for producing a disaccharide comprising the step of contacting a culture of the strain of the present invention or a purified product of the culture with xylan. This is based on experimental results showing that the strain of the present invention can decompose xylan and produce disaccharide as the main product, and that this activity can be exerted in a culture medium from which the bacteria have been removed.

Details regarding culture and purification in the disaccharide production method of the present invention can be referred to the information described above.

For matters regarding contact between a culture of the strain of the present invention or a purified product of the culture and xylan, reference may be made to the description of contact between a culture of the strain of the present invention or a purified product of the culture and polysaccharide.

The disaccharide production method of the present invention may further include the step of purifying the disaccharide produced after contacting it with xylan. At this time, for purification matters, refer to the usual method of purifying a specific reaction product from the reaction mixture after the enzymatic reaction of polysaccharide.

Based on the above characteristics of the strain of the present invention, the present invention also provides a composition for producing disaccharides comprising a culture of the strain of the present invention or a purified product of the culture.

Details regarding culture and purification in the composition for producing disaccharides of the present invention can be referred to the information described above.

The composition for producing disaccharides of the present invention may further include other ingredients in addition to the culture or purification. For example, ingredients that do not substantially inhibit the decomposition activity of xylan by the culture or purification may further include buffers, diluents, excipients, etc. for stable action of the glycolytic enzyme.

The composition for producing a disaccharide of the present invention may include, for example, 0.001 to 100% of the culture or purified product based on the total weight of the composition.

Hereinafter, the present invention will be described in more detail through examples. Since these examples are merely for illustrating the present invention, the scope of the present invention is not to be construed as limited by these examples.

[Example]

1. Method

1-1. Strain isolation

One rose flower collected from the Seo-gu area of Incheon was sterilized by soaking it in 200 ml of 80% ethanol for 10 seconds and washed four times in 500 ml of sterilized water. 50 ㎖ of sterilized water was added to the sample from which water was removed, and the sample was thoroughly crushed in a mortar. 200 ㎕ of the supernatant was spread on MRS solid medium and cultured at 25°C for 2 days to obtain the resulting colonies. A total of 8 colonies were obtained, and all 8 colonies were morphologically classified as fungi (presumed to be yeast).

Eight colonies were transferred to PDA solid medium containing 0.2% AZCL (Azurine-crosslinked)-xylan and cultured at 25°C for 5 days. Decomposition activity was confirmed from the blue color, resulting in one highly active strain. was finally selected.

The final selected strain was named RN2.

1-2. Strain identification

To identify RN2, the ITS region was sequenced using primers ITS1F (5'-CTTGGTCATTTAGAGGAAGTAA-3') and ITS4 (5'-TCCTCCGCTTATTGATATGC-3') and NL1 (5'-GCATATCAATAAGCGGAGGAAAAG-3'). And the nucleotide sequence of the D1D2 region was analyzed using primers NL4 (5'-GGTCCGTGTTTCAAGACGG-3').

Based on the nucleotide sequences of the ITS region and the D1D2 region, homology analysis of the nucleotide sequence was performed from the GenBank database information using the BlastN program of the National Center for Biotechnology Information (NCBI). For phylogenetic tree analysis, the ITS-D1D2 sequences of strains showing homology were obtained, and a NJ (Neighbor-Joining) phylogenetic tree was created using the Mega X program to analyze phylogenetic relationships.

1-3. Analysis of morphological and physiological characteristics

RN2 was cultured in PDA medium to observe the changing morphology of colonies as they grew on the medium, and on the 7th day of culture, hyphae were observed using an Axio Imager Z2 microscope (Zeiss).

Physiological characteristics were investigated using API 20NE, API 20E, and API ZYM kits (Biomerieux, France) according to the presented method. The bacterial sample was obtained by culturing RN2 in PDA medium for 2 days.

1-4. Polysaccharide degradation activity analysis

To examine the polysaccharide decomposition activity of RN2, RN2 was cultured on PDA medium containing 0.2% AZCL-xylan, -amylose, -arabinoxylan, -cellulose, and -pullulan, respectively. After inoculation, it was cultured at 25°C for 48 hours, and it was observed that the AZCL matrix around the colonies was decomposed and turned blue.

RN2 was inoculated into 200 ml of PDB liquid medium and cultured at 30°C for 10 days, sampling 5 ml of the culture medium every 3 days, and centrifuged at 15,000 rpm for 30 minutes to remove only the supernatant. The prepared supernatant was filtered through a 0.22㎛ syringe filter and used as an enzyme solution, and xylanase activity was measured using the DNS (dinitrosalicylic acid) method [Miller GL. 1959. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal. Chem. 31: 426-428.], and the amount of reducing sugar liberated by the enzyme reaction was measured at an absorbance of 540 nm.

1-5. TLC analysis

To analyze the hydrolysis products produced by RN2 by decomposing xylan, samples from the 3rd day, which showed the highest activity as a result of measuring enzyme activity by bacterial growth period, were filtered through an Amicon ultracentrifugal filter (10KDa cut-off, Millipore, USA) was used to prepare a 10-fold concentrated crude enzyme solution.

To analyze xylan degradation products, 100 ㎕ of crude enzyme solution was added to 1X phosphate buffered saline (PBS) containing 0.4% xylan and reacted at 37°C for 24 hours. During the enzyme reaction, the enzyme reaction solution was sampled at 12-hour intervals and 20 μl was added to a silica gel 60 plate (Merck). At this time, xylobiose, xylotetraose, and xylohexaose (Megazyme, Ireland) were used as standards. A solution of n-butanol: acetic acid: distilled water (2:1:2) was used as a developing solvent, and a solution of ethanol: sulfuric acid (9:1) was used as a color developing reagent. After development, the color development reagent was evenly sprinkled and confirmed by heating at 120°C.

2. Results

2-1. Isolation and identification of RN2

RN2 was finally selected as a strain showing xylan decomposition enzyme activity from rose flower samples.

When RN2 was cultured in PDA medium, the central part of the colony was black, but the overall color was light pink. The edges grew in the shape of hyphae, and it was confirmed that the speed of mold cultivation was slower than that of general mold cultivation. Microscopic observation confirmed that it grew in an oval shape (similar to the shape of a typical yeast) (Figure 2).

By analyzing the ITS-D1D2 sequence of RN2, it was classified as a species in the genus Cryptocline . As a result of NCBI Blast homology analysis, RN2 showed the highest homology of 99.83% with Cryptocline arctostaphyli (Table 1). In addition , ITS- As a result of constructing a Neighbor-Joining (NJ) phylogenetic tree based on the D1D2 sequence, it formed the same phylogenetic branch as Cryptocline Arctostaphylli and was judged to be the same species (Figure 1). Based on the ITS-D1D2 sequence homology search results and the NJ phylogenetic tree construction results, RN2 was identified as Cryptocline Arctostaphylli. Finally, the strain RN2 discovered in the present invention was named Cryptocline Arctostaphylli RN2.

ITS-D1D2 base sequence homology search results of RN2 scientific name isolate ITS sequence registration number sameness(%) D1D2 sequence registration number sameness(%) Cryptocline arctostaphyli 19GCAS004 MW077685 98.77 MW077311 99.83 Cryptocline arctostaphyli 19GCAS003 MW077684 98.77 MW077310 99.83 Cryptocline arctostaphyli CBS 454.84 MH861759 98.07 MH873458 99.66 Saccothecium rubi MFLUCC 14-1171 NR_148096 96.32 NG_059644 96.62 Saccothecium sepincola CBS 749.71 MH860331 94.66 MH860331 - Pringsheimia chamaecyparidis CBS 746.71 MH860328 95.81 MH872083 97.13 Pringsheimia smilacis CBS 873.71 AJ244257 96.41 FJ150970 93.02 Pseudosydowia eucalypti CPC:14028 GQ303296 93.35 GQ303327 97.29 Pseudosydowia eucalypti CPC:14927 GQ303297 93.15 GQ303328 97.44 Pseudosydowia eucalypti CBS 131832 MH865934 93.85 MH877368 96.95 Dothichiza pityophila CBS 215.50 AJ244242 95.43 FJ150968 92.18 Kabatiella microsticta CBS 342.66 EU167608 88.62 EU167608 96.46 Aureobasidium pullulans 7R-1-F03 KX958050 89.02 KX958050 96.46 Aureobasidium pullulans 7R-1-F01 KX958048 88.52 KX958048 96.46

Cryptocline Arctostaphylli RN2 was measured for 19 enzyme activities using the API ZYM kit, and the results showed alkaline phosphatase, esterase (C4), and esterase lipase. ) (C8), lipase (C14), leucine arylamidase, valine arylamidase, cystine arylamidase, trypsin, alpha- α-chymotrypsin, acid phosphatase, Naphthol-AS-BI-phosphohydrolase, α-galactosidase, Beta-galactosidase, beta-glucuronidase, alpha-glucosidase, beta-glucosidase, N-acetyl -The activity of beta-glucosamanidase (N-acetyl-β-glucosaminidase) was positive, and the other two enzyme activities were negative. As a result of measuring enzyme activity and fermentation reaction using the API 20E kit, beta-galactosidase, urease, and tryptophane deaminase enzyme activities were positive, and other enzyme reactions were negative. And the fermentation test results for sugars were positive for D-glucose, D-sucrose, D-melibiose, and amygdalin, and the remaining five types of sugars were fermented. was not observed. In the API 20NE kit test, urease, esculin degradation, beta-galactosidase enzyme activity, and glucose fermentation reaction were positive, and D-glucose, L-arabinose, and D-mannose as carbon sources. (D-mannose), D-mannitol (D-mannitol), D-maltose (D-maltose), and potassium gluconate showed positive reactions. Other reactions related to the use of enzymes and carbon sources were negative.

API ZYM kit analysis results of cryptocline Arctostaphylli RN2 enzyme reaction Alkaline phosphatase + Esterase(C4) + Esterase Lipase(C8) + Lipase(C14) + Leucine arylamidase + Valine arylamidase + Cystine arylamidase + Trypsin +- α-chymotrypsin +- Acid phosphatase + Naphtol-AS-BI-phosphohydrolase + α-galactosidase + β-galactosidase + β-glucuronidase +- α-glucosidase + β-glucosidase + N-acetyl-D-β-glucosaminidase + α-mannosidase - α-fucosidase -

Symbol: +, positive; +-, weakly positive; -, voice.

API 20E kit analysis results of cryptocline Arctostaphylli RN2 Enzyme/Substrate reaction enzyme β-galactosidase +- Arginine dihydrolase - Lysine decarboxylase - Ornithine decarboxylase - Citrate utilization - H₂S production - Urease + Tryptophane deaminase + indole production - acetoin production
(Vogesproskauer) - Gelatinase - fermentation D-glucose + D-mannitol - inositol - D-sorbitol - L-rhamnose - D-sucrose + D-melibiose + amygdalin -+ L-arabinose -

Symbol: +, positive; +-, weakly positive; -, voice.

API 20NE kit analysis results of cryptocline Arctostaphylli RN2 Enzyme/Substrate reaction enzyme Nitrates to nitrites - Indole production - Arginine dihydrolase - Urease +- Esculin hydrolysis + Gelatin hydrolysis - Galactosidase +- fermentation Glucose +- Fairytale D-glucose + L-arabinose + D-mannose + D-mannitol +- N-acetyl-glucosamine - D-maltose +- potassium gluconate +- capric acid - adipic acid - malic acid - trisodium citrate - phenylacetic acid -

Symbol: +, positive; +-, weakly positive; -, voice.

2-2. Polysaccharide decomposition activity

Cryptocline Arctostaphili RN2 decomposes insoluble AZCL-amylose, AZCL-arabinoxylan, and AZCL-xylan around the cell, and appears blue in the medium, showing that it secretes decomposing enzymes to the outside of the cell. Among the three types of enzyme activities, xylan decomposition activity was observed to be the strongest, followed by arabinoxylan decomposition activity, followed by amylose decomposition activity (Figure 3).

Since decomposition activity for AZCL-xyloglucan, -cellulose, and -pullulan was not observed, it is assumed that xyloglucanase, cellulase, and pullulanase enzymes are not produced. .

When measuring xylanase activity according to liquid culture of Cryptocline Arctostaphylli RN2 in PDB medium, the highest enzyme activity was seen in the sample on the 3rd day, but rapidly decreased from the 5th day (Figure 4).

When the enzyme reaction product using xylan as a substrate was analyzed by TLC using the crude enzyme solution prepared from the sample of the 3rd day of liquid culture of Cryptocline Arctostaphylli RN2, the xylanase secreted by Cryptocline Arctostaphylli RN2 was not present. It was found that beechwood xylan was decomposed to produce dimer (DP2) as the main decomposition product (FIG. 5). It was confirmed that oligosaccharides with a structure longer than the hexamer (DP6) were produced, including the dimer, which is the final decomposition product, and this TLC pattern is consistent with the hydrolysis characteristics of a general endo-type xylanase. Therefore, it is believed that by using the enzyme of the cryptocline Arctostaphylli RN2, it is possible to produce xylooligosaccharide, which uses dimers as the main product from xylan.

Agricultural Biotechnology Research Institute KACC93360P 20210615

<110> National Institute of Biological Resources <120> Novel strain Cryptocline arctostaphyli RN2 having xylan degrading activity <130> ALP21012 <160> 2 <170> KoPatentIn 3.0 <210> 1 <211> 569 <212> DNA <213> Unknown <220> <223> Cryptocline arctostaphyli RN2 <400> 1 cggaaggatc attaaagaga aaggggctaa acaccccgac ctccaaccct ctgttgttaa 60 aactactttg ttgctttggc gggaccgttc ggtctccgag ccgcccaggc accttcacgg 120 gtgtccaggc gagtgcccgc cagagttaaa ccaaactctt gttatttaaa ccggtcgtct 180 gagtacaaat ttttgaatta aatcaaaact ttcaacaacg gatctcttgg ttctcgcatc 240 gatgaagaac gcagcgaaat gcgataagta atgtgaattg cagaattcag tgaatcatcg 300 aatctttgaa cgcacattgc gccccttggt attccgaggg gcatgcctgt tcgagcgtca 360 ttacaccact caagcatagc ttggtattgg gcaatcgccc ccgccagtcg gggacgcgcc 420 tcaaacacct cggcggagcc tcaccggctt tgggcgtagt agaatttcta ataacgtcct 480 ttaacggaga tggttccttt gccgacagaa gcctttaaat ttttctaggt tgacctcgga 540 tcaggtaggg atacccgctg aacttaagc 569 <210> 2 <211> 591 <212> DNA <213> Unknown <220> <223> Cryptocline arctostaphyli RN2 <400> 2 aggaaaaagaa accaacaggg attgccctag taacggcgag tgaagcggca acagctcaaa 60 tttgaaagct agccttcggg ttcgcattgt aatttgtaga ggatgctttg gggcagccgc 120 ctgtctaagt tccttggaac aggacgtcat agagggtgag aatcccgtat gtgacaggac 180 atggcaccct atgtaaagct ccttcgacga gtcgagttgt ttgggaatgc agctctaaat 240 gggaggtaaa tttcttctaa agctaaatac cggcgagaga ccgatagcgc acaagtagag 300 tgatcgaaag atgaaaagca ctttggaaag agagttaaaa agcacgtgaa attgttgaaa 360 gggaagcgct tgcaatcaga cttgtcttta actgttcggc cggtcttctg accggtttac 420 tcagtaatcg acaggccagc atcagtttcg gcggtcggat aaaggccctg ggaatgtggc 480 tctgccttcg ggtagagtgt tatagcccag ggtgtaatac ggccagccgg gactgaggtc 540 cgcgattcgt ctaggatgct ggcgtaatgg ttgtaagcga cccgtcttga a 591

Claims (7)

Cryptocline arctostaphyli RN2 strain deposited with accession number KACC 93360P. A method of decomposing xylan, arabinoxylan, or amylose comprising contacting a culture of the strain of claim 1 or a purified product of the culture with a polysaccharide. delete A composition for decomposing xylan, arabinoxylan, or amylose comprising a culture of the strain of claim 1 or a purified product of the culture. delete A disaccharide production method comprising the step of contacting a culture of the strain of claim 1 or a purified product of the culture with xylan. A composition for producing a disaccharide comprising a culture of the strain of claim 1 or a purified product of the culture.