KR101082030B1 - Method for preparing (-)-gamma-lactam and novel microbial strains for the same - Google Patents

Abstract

The present invention relates to a novel microbial strain and a method for preparing (-)-gamma-lactam ((-)-2-azabicyclo [2.2.1] hept-5-en-3-one) using the same. Preferably, the novel microbial strain is reacted with racemic gamma-lactam to selectively hydrolyze only (+)-gamma-lactam, thereby producing (-)-gamma-lactam having a very high optical purity of 99% ee or more. It relates to a manufacturing method. The microbial strains used in the present invention are new microorganisms not mentioned in the prior art, and are very useful industrially because they are easy to grow and cultivate and can produce (-)-gamma-lactam having high optical purity with high yield in a short time. Do.

Gamma lactam, (-)-2-azabicyclo [2.2.1] hept-5-en-3-one, Rhodococcus, Pseudomonas, Aureobacterium

Description

Method for preparing (-)-gamma-lactam and novel microbial strains used therein {Method for preparing (-)-gamma-lactam and novel microbial strains for the same}

1 is an isolated strain Acromobacter xylose oxidase subspecies of the present invention. Xylosoxidance SKM01 ( Achromobacter xylosoxidans subsp. Xylosoxidans SKM01 (KCCM-10547)) shows the identification of the taxonomic map.

Figure 2 shows the taxonomic map found as a result of the identification of the isolated strain paracoccus aminoborans SKM02 (KCCM-10548) of the present invention.

Figure 3 shows the taxonomic map found by the identification of the isolated strain Pseudomonas aeruginosa SKM07 (KCCM-10549) of the present invention.

The present invention provides a novel microbial strain and an optically active (-)-2-azabicyclo [2.2.1] hept-5-en-3-one ((-)-2-azabicyclo [2,2,1] hept -5-en-3-one, hereinafter referred to as '(-)-gamma-lactam' (Formula 1)), more specifically gamma of racemic body using a novel microbial strain- The present invention relates to a method of optically hydrolyzing only (+)-gamma-lactam in lactam to produce (-)-gamma-lactam having very high optical purity of 99% or more.

(-)-Gamma-lactam referred to in the present invention is used as an intermediate required for the synthesis of carbocyclic nucleosides such as Abacavir, Carbovir, etc., which are used as a therapeutic agent for viral diseases such as AIDS. As the demand for gamma-lactam of optical activity is continuously increasing, many attempts have been made to prepare the compound effectively. An enzymatic resolution method mainly using enzymes has been studied. Enzymatic optical splitting of gamma-lactam using lactamase-producing microorganisms developed by Chiroscience, UK.

In 1990, Enzymatix, the forerunner of Chiroscience, announced a method of optically splitting gamma-lactams using two new microbes that produce an enzyme called lactamase. In this method, Rhodococcus sp. ENZA-1 (NCIMB 40213) hydrolyzes (-)-gamma-lactam and finally (+)-gamma-lactam and (-)- Produce amino acids In addition, other microorganisms, Pseudomonas solanacearum ENZA-20 (NCIMB 40249), on the other hand, hydrolyze (+)-gamma-lactams, resulting in (-)-gamma-lactams and (+)-amino acids. . However, this method takes about 14 days to enzymatic reaction, and in particular, ENZA-1 has a disadvantage of being difficult to apply commercially since the optical purity of the enzymatic reaction product (-)-amino acid is 81% ee. EP 0424064, USP 5284769, USP 5498625).

In 1993 Chiroscience and Exter University jointly published an optical splitting method of gamma-lactam using two new microorganisms different from the microorganisms mentioned in the patent. The microorganisms used in this method are Pseudomonas fluorescens ENZA-22 and Aureobacterium sp. ENZA-25 . In the former case, (+)-gamma-lactam is hydrolyzed to give final results. With (-)-gamma-lactam and (+)-amino acid. In the latter case, hydrolysis of (-)-gamma-lactam results in (+)-gamma-lactam and (-)-amino acid. . Even in this case, and the optical purity of the desired optically active compound with low downside 93%, 91% ee, respectively, there is also a case of Pseudomonas Solana serum ENZA-22 is a very unstable enzyme disadvantages (Tetrahedron: Asymmetry Vol.4 , No. 6, pp. 1117-1128, 1993).

In 1998 Chiroscience published an optical splitting method of gamma-lactam using new microorganisms with improved thermal and substrate stability compared to previous microorganisms in WO 98/10075. The microorganism used at this time is Comamonas acidovorans (NCIMB 40827), which hydrolyzes (+)-gamma-lactam to finally produce (-)-gamma-lactam and (+)-amino acid. Optical microfractionation of racemic gamma-lactam using the microbial strain obtained through fermentation resulted in about 44% yield of (-)-gamma-lactam with an optical purity of 99% ee or more after 24 hours reaction. It was successfully used to commercialize the (-)-gamma-lactam production process.

Unlike the above-mentioned method, the method of making optically active (-)-gamma-lactam by selectively hydrolyzing by using a hydrolysis enzyme after introducing hydroxymethyl group or acetyl group into amide group of gamma-lactam It has been reported. In 1996, Hiroto Nakano et al. Published a method for optically dividing N-hydroxymethyl derivatives of gamma-lactams using a hydrolase called lipase PS from Amano. However, this method requires a total of four steps including two enzymatic reactions in order to obtain pure (-)-gamma-lactam with an optical purity of 99% ee or more, and the overall yield is very low, about 14%. There are difficult drawbacks (Tetrahedron: Asymmetry, Vol. 7, No. 8, pp. 2381-2386, 1996).

In 2002, SmithKline Beecham reported that racemic t-butyl 3-oxo-2-azabicyclo [2.2.1] hept-5-ene-carboxylate or racemic cis-2-acetyl-2-azabicyclo [2.2.1] A method of optically dividing a hept-5-en-3-one derivative using a hydrolase called Savinase from Novo (USP 6340787) has been published. In this method, (-)-gamma-lactam derivatives having an optical purity of 99% ee or more can be prepared using enzymes. However, the amount of enzyme used is large and the reaction time is about two days longer than that of the substrate. The total yield drops to 20% during the three-step reaction to make gamma-lactam, which is also difficult to commercialize.

As a result of examining the prior arts as described above, in order to commercialize the method of preparing (-)-gamma-lactam useful for the synthesis of carboxynucleosides used as a therapeutic agent for viral diseases, it is possible to use gamma- using microorganisms or enzymes. It is believed that a method of directly hydrolyzing the lactam itself is necessary.

It is an object of the present invention to provide a new method for producing (-)-gamma-lactam having a very high optical purity of 99% or more using this novel microbial strain.

In order to achieve the above object, the method of the present invention adds racemic gamma-lactam to sodium phosphate buffer solution and performs a selective hydrolysis reaction using a novel microorganism found in the present invention. ) -Gamma-lactam.

The present inventors collected microorganisms directly from the surrounding environment to find microorganisms that selectively hydrolyze only (+)-gamma-lactam, and the three microorganisms having excellent performance as a result of performing a hydrolysis reaction on the collected microorganisms Could find. The newly discovered microorganisms were identified through the Korean Culture Center of Microorganisms (KCCM) and all three microorganisms were identified as new microorganisms not mentioned in the prior art. Therefore, the present inventors have developed a method for finally producing optically active (-)-gamma-lactam by selectively hydrolyzing only (+)-gamma-lactam in racemic gamma-lactam using a novel microorganism found by itself. The present invention has been completed based on this.

Hereinafter, the present invention will be described in more detail.

The present invention relates to a method for preparing (-)-gamma-lactam using a microbial strain, which is a new microorganism not yet mentioned in the prior art Acromobacter xylose oxidase subspecies. Optical selection of (+)-gamma-lactam only using strains such as Achromobacter xylosoxidans subsp.Xylosoxidans , Paracoccus aminovorans and Pseudomonas aeruginosa Decomposition provides (-)-gamma-lactam with very high optical purity of 99% ee or more.

The selective hydrolysis process according to the invention is as follows.

(1): 2-azabicyclo [2.2.1] hept-5-en-3-one (gamma-lactam)

(2): 4-amino-cyclopent-2-ene carboxylic acid

In the present invention, the enzyme reaction temperature is preferably 20-50 ° C., particularly, the enzyme activity is maintained even at a temperature of 40 ° C. or higher, and thus, the temperature stability was excellent. In addition, the pH of the enzyme reaction was found to be in the proper range of pH 4-10, it was found to be excellent microorganisms with excellent pH stability as there is almost no decrease in activity even with the change of pH.

Hereinafter, the present invention will be described in more detail with reference to the following examples, but the present invention is not limited to the following examples.

Example 1 (Secure Microbial Strains)

In order to obtain a microbial strain that selectively hydrolyzes only (+)-gamma-lactam, samples were collected from soil and wastewater of Gapcheon (Daejeon, South Korea). After adding the buffer solution PBS (Phosphate-buffered saline) to the sample was stirred and left to stand. After the precipitation was completed, the supernatant was plated on a solid medium containing only N-acetyl-L-phenylalanine as the only carbon source and incubated for 7 days at 30 ° C. After 7 days to obtain a strain showing the growth it was grown in each nutrient medium to obtain the cells. 100 mM phosphate buffer solution (pH7.0) containing 1% (W / V) racemic gamma-lactam was added to the obtained cells to carry out the reaction for 3 days, and after completion of the reaction, the strain was separated using a centrifuge. After the supernatant was extracted three times using the same amount of ethyl acetate. The extract was distilled under reduced pressure to remove ethyl acetate to obtain a gamma-lactam product. The secured product was analyzed using liquid chromatography.

The analysis conditions of the liquid chromatography are as follows.

As a column, chiralcel OD-H made of cellulose carbamate derivative (chiralcel OD-H, Daicel Co., Ltd.) was used. As eluent, hexane, isopropanol and trifluoroacetic acid were used at a ratio of 95: 5: 0.1. The mixed solution was flowed at a rate of 1 ml per minute, and the reaction product was detected at UV 205 nm. The reactants and products of the reaction, (+)-gamma-lactams, are readily distinguishable since they are detected at 13.6 minutes and (-)-gamma-lactams at 21.0 minutes.

The above process was repeated to selectively hydrolyze only (+)-gamma-lactam to secure three strains that finally leave (-)-gamma-lactam, and three species were identified.                     

Example 2 (Identification of Microbial Strains)

Three strains isolated in Example 1 were named SKM01, SKM02, and SKM07, respectively, and were identified by requesting them to the Korean Culture Center of Microorganisms (KCCM). For reference, 16S rDNA sequence analysis results of the SKM01, SKM02, and SKM07 strains are shown in SEQ ID NOs: 1 to 3, and the physiological analysis results are shown in Table 1 below. In addition, the taxonomic maps identified as a result of the identification are shown in FIGS. 1, 2, and 3, respectively. As a result, SKM01 is acromobacter xylose redox subspecies. Achromobacter xylosoxidans subsp.Xylosoxidans , SKM02 for Paracoccus aminovorans and SKM07 for Pseudomonas aeruginosa, all three strains have been identified in the literature Turned out to be another new strain. These strains were deposited to the Korea microbial conservation center as of December 3, 2003, and their accession numbers are KCCM-10547, KCCM-10548, and KCCM-10549, respectively.

Physiological Characteristics of the Strains of the Invention Analysis item SKM01 SKM02 SKM07 Reduction of Nitrate to Nitrite + + - Indole generation - - - Glucose acidification - - - Arginine Dehydrolase + - + Urease + - + Esculin hydrolysis (β-glucosidase) - - - Gelatin Hydrolysis (Protease) - - + β-galactosidase - - - Glucose assimilation - + + Arabian fairy tale - + - Mannose Fairy Tale - + - Mannitol fairy tale - + + N-acetyl-glucosamine assimilation - - + Maltose fairy tale - + - Gluconate assimilation + - + Caprate Fairy Tale - - + Adipate fairy tale + - + Malate fairy tale + + + Citrate Fairy Tale + - + Phenyl-acetate assimilation + - - Cytochrome oxidizer + + -

Example 3 (Segmentation of Racemic Gamma-lactams Using Microbial Strains)

0.5 g of the microorganism strain SKM01 of Example 2 and 0.2 g of racemic gamma-lactam were quantitatively added to the reactor, followed by addition of 100 mM phosphate buffer solution (pH7.0) to adjust the final volume to 10 ml, followed by reaction at 30 ° C. Was performed. After the reaction, a certain amount of contents were collected at a predetermined time to obtain (-)-gamma-lactam in the same manner as in Example 1. The optical purity of the thus prepared (-)-gamma-lactam was analyzed using liquid chromatography in the same manner as in Example 1.

The results of the analysis were expressed as yield [%] and optical purity [% ee]. The yield of (-)-gamma-lactam was calculated using Equation 1 below, and the optical purity of (-)-gamma-lactam was Calculated using 2.

The optical purity [% ee] of the (-)-gamma-lactam according to the reaction time under the above conditions is shown in Table 2 below.

Optical purity of (-)-gamma-lactam according to reaction time [% e.e.] Response time (hours) Optical purity (% e.e.) 4 77.7 8 94.8 12 98.8 16 99.7 20 99.9

After 20 hours, the reaction was stopped, transferred to a separatory funnel, extracted three times with 10 mL of ethyl acetate, and the organic solvent layer was separated and the solvent was removed by distillation under reduced pressure to recover 0.0798 g of (-)-gamma-lactam. This was analyzed using liquid chromatography. At this time, the yield of (-)-gamma-lactam was 39.9%, the optical purity was 99.9% ee.

Examples 4-6

In Example 3, the strain amount, pH of the buffer solution, and the reaction temperature were changed to 0.1, 0.5, and 1 g of the mass under the same conditions as in Example 3, followed by reaction for 20 hours, and then the reactants were prepared in the same manner as in Example 3. The recovered and analyzed results are shown in Table 3.

Yield [%] and optical purity [% e.e.] Of (-)-gamma-lactam according to the change of basis mass (g) Example Mass (g) yield[%] Optical purity [% e.e.] 4 0.1 38.2 99.9 5 0.5 40.5 99.9 6 One 36.6 99.8

Examples 7-8

Among the conditions of Example 3, the mass, the pH of the buffer solution, and the reaction temperature were changed to 0.05, 0.1, and 0.2 g of the strain under the same conditions as in Example 3, and then reacted for 20 hours, in the same manner as in Example 3. The results of the recovery and analysis of the reactants are shown in Table 4.

Yield [%] and optical purity [% e.e.] Of (-)-gamma-lactam according to strain (g) change Example Strain amount (g) yield[%] Optical purity [% e.e.] 7 0.1 42.0 99.7 8 0.2 40.7 99.9

Examples 9-14

The amount of substrate and strain, the reaction temperature, and the volume of the buffer solution in the conditions of Example 3 were varied by changing the pH of the buffer solution to 4, 5, 6, 8, 9, 10 under the same conditions as in Example 3. After the reaction for a time, the reaction was recovered and analyzed in the same manner as in Example 3 is shown in Table 5.

Yield [%] and optical purity [% e.e.] Of (-)-gamma-lactam according to pH change of buffer solution Example PH of buffer yield[%] Optical purity [% e.e.] 9 4 41.1 99.9 10 5 40.8 99.9 11 6 40.7 99.8 12 8 39.5 99.8 13 9 39.4 99.9 14 10 38.1 99.9

Examples 15-19

In the conditions of Example 3, the amount of substrate and strain, pH and volume of the buffer solution were changed for 25 hours to 25, 35, 40, 45, and 50 ° C under the same conditions as in Example 3, followed by reaction for 20 hours. In Table 6, the reaction was recovered and analyzed in the same manner as in Example 3.                     

Yield [%] and optical purity [% e.e.] Of (-)-gamma-lactam according to reaction temperature (° C) Example Temperature (℃) yield[%] Optical purity [% e.e.] 15 25 39.0 99.0 16 35 37.8 99.8 17 40 40.8 99.7 18 45 38.8 99.8 19 50 36.2 99.9

Examples 20-21

PH, volume, base mass, and reaction temperature of the buffer solution in Example 3 were changed to the types of microbial strains SKM02 and SKM07 under the same conditions as in Example 3, followed by reaction for 3 days, and then the same as in Example 3. The reaction was recovered and analyzed by the method shown in Table 7.

Yield [%] and optical purity [% e.e.] Of (-)-gamma-lactam according to the microorganism strain Example Strain name yield[%] Optical purity [% e.e.] 20 SKM02 30.0 99.3 21 SKM07 15.2 99.4

As can be seen from the above examples, the microbial strains used in the present invention are new microorganisms not mentioned in the prior art, and when these microorganisms are used to produce (-)-gamma-lactam, they have a high optical purity of 99% ee or more. The lactam compound can be prepared, and especially when SKM01 strain is used, a high yield of 40% or more can be achieved, which is commercially useful. In addition, since the reaction proceeds at room temperature and normal pressure, there is no need for an additional device, and there is an advantage in that a process suitable for mass production can be developed due to easy cultivation of microorganisms.

<110> SK CORPORATION <120> Method for preparing (-)-gamma-lactam using novel microbial          strains <160> 3 <170> KopatentIn 1.71 <210> 1 <211> 1440 <212> DNA <213> Achromobacter xylosoxidans subsp. xylosoxidans <400> 1 tgacgctagc gggatgcctt acacatgcaa gtcgaacggc agcacggaac ttcggtctgg 60 tggcgagtgg cgaacgggtg agtaatgtat cggaacgtgc ccagtagcgg gggataacta 120 cgcgaaagcg tagctaatac cgcatacgcc ctacggggga aagcagggga tcgcaagacc 180 ttgcactatt ggagcggccg atatcggatt agctagttgg tggggtaacg gctcaccaag 240 gcgacgatcc gtagctggtt tgagaggacg accagccaca ctgggactgn agnacacggc 300 ccagactcct acgggaggca gcagtgggga attttggaca atgggggaaa ccctgatcca 360 gccatcccgc gtgtgcgatg aaggccttcg ggttgtaaag cacttttggc aggaaagaaa 420 cgtcgcgggt taataccccg cggaactgac ggtacctgca gaataagcac cggctaacta 480 cgtgccagca gccgcggtaa tacgtagggt gcaagcgtta atcggaatta ctgggcgtaa 540 agcgtgcgca ggcggttcgg aaagaaagat gtgaaatccc agagcttaac tttggaactg 600 catttttaac taccgggcta gagtgtgtca gagggaggtg gaattccgcg tgtagcagtg 660 aaatgcgtag atatgcggag gaacaccgat ggcgaaggca gcctcctggg ataacacgac 720 gctcatgcac gaaagcgtgg ggagcaaaca ggattagata ccctggtagt ccacgcccta 780 aacgatgtca actagctgtt ggggtcttcg gaccttggta gcgcagctaa cgcgtgaagt 840 tgaccgcctg gggagtacgg tcgcaagatt aaaactcaaa ggaattgacg gggacccgca 900 caagcggtgg atgatgtgga ttaattcgat gcaacgcgaa aaaccttacc tacccttgac 960 atgtctggaa tgccgaagag atttggcagt gctcgcaaga gaactggaac acaggtgctg 1020 catggctgtc gtcagctcgt gtcgtgagat gttgggttaa gtcccgcaac gagcgcaacc 1080 cttgtcatta gttgctacga aagggcactc taatgagact gccggtgaca aaccggagga 1140 aggtggggat gacgtcaagt cctcatggcc cttatgggta gggcttcaca cgtcatacaa 1200 tggtcgggac agagggtcgc caacccgcga gggggagcca atcccagaaa cccgatcgta 1260 gtccggatcg cagtctgcaa ctcgactgcg tgaagtcgga atcgctagta atcgcggatc 1320 agcatgtcgc ggtgaatacg ttcccgggtc ttgtacacac cgcccgtcac accatgggag 1380 tgggttttac cagaagtagt tagcctaacc gcaagggggg cgattaccac ggtaggattc 1440                                                                         1440 <210> 2 <211> 1372 <212> DNA <213> Paracoccus aminovorans <400> 2 gctggcggca ggcctaacac atggcaagtc gagcgagatc ttcggatcta gcggcggacg 60 ggtgagtaac gcgtgggaat atgcccttct ctacggaata gcctcgggaa actgggagta 120 ataccgtata cgcccttagg gggaaagatt tatcggagaa ggattagccc gcgttggatt 180 aggtagttgg tggggtaatg gcctaccaag ccgacgatcc atagctggtt tgagaggatg 240 atcagccaca ctgggactga gacacggccc agactcctac gggaggcagc agtggggaat 300 cttagacaat gggggcaacc ctgatctagc catgccgcgt gagtgatgaa ggccttaggg 360 ttgtaaagct ctttcagctg ggaagataat gacggtacca gcagaagaag ccccggctaa 420 ctccgtgcca gcagccgcgg taatacggag ggggctagcg ttgttcggaa ttactgggcg 480 taaagcgcac gtaggcggac cagaaagttg gaggtgaaat cccagggctc aaccttggaa 540 ctgccttcaa aactattggt ctggagttcg agagaggtga gtggaattcc gagtgtagag 600 gtgaaattcg tagatattcg gaggaacacc agtggcgaag gcggctcact ggctcgatac 660 tgacgctgag gtgcgaaagc gtggggagca aacaggatta gataccctgg tagtccacgc 720 cgtaaacgat gaatgccagt cgtcgggtag catgctattc ggtgacacac ctaacggatt 780 aagcattccg cctggggagt acggtcgcaa gattaaaact caaaggaatt gacgggggcc 840 cgcacaagcg gtggagcatg tggtttaatt cgaagcaacg cgcagaacct taccaaccct 900 tgacattaca ggacatcccc agagatgggg ctttcacttc ggtgacctgt ggacaggtgc 960 tgcatggctg tcgtcagctc gtgtcgtgag atgttcggtt aagtccggca acgagcgcaa 1020 cccacactct tagttgccag cattcagttg ggcactctaa gagaactgcc gatgataagt 1080 cggaggaagg tgtggatgac gtcaagtcct catggccctt acgggttggg ctacacacgt 1140 gctacaatgg tggtgacagt gggttaatcc ccaaaagcca tctcagttcg gattggggtc 1200 tgcaactcga ccccatgaag ttggaatcgc tagtaatcgc ggaacagcat gccgcggtga 1260 atacgttccc gggccttgta cacaccgccc gtcacaccat gggagttggt tctacccgac 1320 ggccgtgcgc taaccagcaa tggaggcagc ggaccacggt aggatcagcg ac 1372 <210> 3 <211> 1447 <212> DNA <213> Pseudomonas aeruginosa <400> 3 tgacgctggc ggcaggccta acacatgcaa gtcgagcgga tgaagggagc ttgctcctgg 60 attcagcggc ggacgggtga gtaatgccta ggaatctgcc tggtagtggg ggataacgtc 120 cggaaacggg cgctaatacc gcatacgtcc tgagggagaa agtgggggat cttcggacct 180 cacgctatca gatgagccta ggtcggatta gctagttggt ggggtaaagg cctaccaagg 240 cgacgatccg taactggtct gagaggatga tcagtcacac tggaactgag acacggtcca 300 gactcctacg ggaggcagca gtggggaata ttggacaatg ggcgaaagcc tgatccagcc 360 atgccgcgtg tgtgaagaag gtcttcggat tgtaaagcac tttaagttgg gaggaagggc 420 agtaagttaa taccttgctg ttttgacgtt accaacagaa taagcaccgg ctaacttcgt 480 gccagcagcc gcggtaatac gaagggtgca agcgttaatc ggaattactg ggcgtaaagc 540 gcgcgtaggt ggttcagcaa gttggatgtg aaatccccgg gctcaacctg ggaactgcat 600 ccaaaactac tgagctagag tacggtagag ggtggtggaa tttcctgtgt agcggtgaaa 660 tgcgtagata taggaaggaa caccagtggc gaaggcgacc acctggactg atactgacac 720 tgaggtgcga aagcgtgggg agcaaacagg attagatacc ctggtagtcc acgccgtaaa 780 cgatgtcgac tagccgttgg gatccttgag atcttagtgg cgcagctaac gcgataagtc 840 gaccgcctgg ggagtacggc cgcaaggtta aaactcaaat gaattgacgg gggcccgcac 900 aagcggtgga gcatgtggtt taattcgaag caacgcgaag aaccttacct ggccttgaca 960 tgctgagaac tttccagaga tggattggtg ccttcgggaa ctcanacaca ggtgctgcat 1020 ggctgtcgtc agctcgtgtc gtgagatgtt gggttaagtc ccgtaacgag cgcaaccctt 1080 gtccttagtt accagcacct cgggtgggca ctctaaggag actgccggtg acaaaccgga 1140 ggaaggtggg gatgacgtca agtcatcatg gcccttacgg ccagggctac acacgtgcta 1200 caatggtcgg tacaaagggt tgccaagccg cgaggtggag ctaatcccat aaaaccgatc 1260 gtagtccgga tcgcagtctg caactcgact gcgtgaagtc ggaatcgcta gtaatcgtga 1320 atcagaatgt cacggtgaat acgttcccgg gccttgtaca caccgcccgt cacaccatgg 1380 gagtgggttg ctccagaagt agctagtcta accgcaaggg ggacggttac cacggagtga 1440 ttcagac 1447

Claims (7)

Acromobacter xylose oxidase subspecies having the property of selectively hydrolyzing (+)-gamma-lactams in racemic gamma-lactams. Xylosoxidance SKM01 ( Achromobacter xylosoxidans subsp. Xylosoxidans SKM01 (KCCM-10547)). delete delete Microbial strain, Acromobacter xylose oxidase subspecies, having the property of selectively hydrolyzing only (+)-gamma-lactam in gamma-lactam. Incubating Xylose oxidance SKM01 ( Achromobacter xylosoxidans subsp. Method for producing (-)-gamma-lactam. delete The method of claim 4, wherein the hydrolysis reaction is performed at pH 4-10. The method of claim 4, wherein the hydrolysis reaction is performed at 20-50 ° C.

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