KR100443652B1 - Actinoplanes teichomyceticus MSL 1510 producing teicoplanin - Google Patents

Actinoplanes teichomyceticus MSL 1510 producing teicoplanin Download PDF

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KR100443652B1
KR100443652B1 KR10-2002-0028359A KR20020028359A KR100443652B1 KR 100443652 B1 KR100443652 B1 KR 100443652B1 KR 20020028359 A KR20020028359 A KR 20020028359A KR 100443652 B1 KR100443652 B1 KR 100443652B1
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김창진
이재찬
박동진
손광희
윤기홍
박용수
권무길
권영일
이민주
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근화제약주식회사
한국생명공학연구원
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Abstract

본 발명은 테이코플라닌(teicoplanin)을 생산하는 야생균주 액티노플라네스 테이코마이세티커스(Actinoplanes teichomyceticus) ATCC 31121의 변이주 액티노플라네스 테이코마이세티커스 MSL 2211 (기탁번호 : KCTC 8926P)를 모균주로 하여 인위적 돌연변이법 및 단일 포자 분리법에 따라 처리하여 모균주의 형질을 개량함으로서 모균주에 비해 테이코플라닌 고생산능을 갖는 변이주 액티노플라네스 테이코마이세티커스 MSL 1510 (기탁번호 : KCTC 10200BP)및 이 신규 변이주를 이용한 테이코플라닌 대량 생산 방법에 관한 것이다.The present invention is a variant strain of actinoplanes teichomyceticus ATCC 31121 producing teicoplanin (teicoplanin) MSL 2211 (Accession Number: KCTC 8926P) Mutant strain Actinoplanes teicomyceticus MSL 1510 having higher yield of teicoplanin compared to the parent strain by treating with the parent strain and by the method of artificial mutation and single spore separation. : KCTC 10200BP) and a method for mass production of teicoplanin using this novel mutant strain.

Description

테이코플라닌 생산 변이주 액티노플라네스 테이코마이세티커스 ＭＳＬ 1510 {Actinoplanes teichomyceticus MSL 1510 producing teicoplanin}Teicoplanin production variant Actinoplanes teicomyceticus MSL 1510 {Actinoplanes teichomyceticus MSL 1510 producing teicoplanin}

본 발명은 테이코플라닌(teicoplanin)을 생산하는 미생물에 관한 것이다. 보다 상세하게는, 본 발명은 야생균주 액티노플라네스 테이코마이세티커스 ATCC31121의 변이주 액티노플라네스 테이코마이세티커스 MSL 2211 (기탁번호 : KCTC 8926P)를 인위적 돌연변이법 및 단일 포자 분리법에 따라 처리하여 모균주에 비해 테이코플라닌 생산능을 향상시킨 변이주에 관한 것이다. 또한, 이 변이주를 이용하여 테이코플라닌을 고수율로 생산하는 방법에 관한 것이다.The present invention relates to a microorganism producing teicoplanin. More specifically, the present invention is a variant strain of wild strain Actinoplanes Teicomyceticus ATCC31121 MSL 2211 (Accession No .: KCTC 8926P) according to the artificial mutagenesis and single spore separation method It relates to a mutant strain that is treated to improve teicoplanin production compared to the parent strain. The present invention also relates to a method for producing teicoplanin in high yield using this mutant strain.

내성 병원균에 대한 대표적인 항생제로는 글라이코펩타이드계열의 항생제가 있으며, 반코마이신 및 테이코플라닌 두 종류가 현재 임상에서 사용되고 있다. 테이코플라닌은 방선균인 액티노플라네스 테이코마이세티커스(Actinoplanes teichomyceticus) 종에 속하는 균주에 의해 생산되는 글라이코펩타이드계 항생제로서, 그람양성 세균 및 메치실린 내성 포도상구균(MRSA) 치료제로 사용되고 있고, 반코마이신에 비해 약효가 우수하고 부작용이 적기 때문에 그 수요가 증가하고 있으며, 국내로의 수입도 매년 큰 폭으로 증가되고 있는 실정이다. 그러나 테이코플라닌은 화학적으로 합성하기가 어려울 뿐만 아니라, 아직까지 생합성 과정이 완전히 규명되지 않아서 발효를 통한 대량 생산방법이 모색되고 있다.Representative antibiotics for resistant pathogens are glycopeptides, and vancomycin and teicoplanin are currently used in clinical practice. Teicoplanin is a glycopeptide antibiotic produced by a strain belonging to actinomycetes Actinoplanes teichomyceticus species, and is used for the treatment of Gram-positive bacteria and methicillin-resistant Staphylococcus aureus (MRSA). In addition, the drug is superior to vancomycin and has fewer side effects, so the demand is increasing, and imports into the country are increasing significantly every year. However, teicoplanin is not only difficult to synthesize chemically, but the biosynthesis process has not been fully identified. Therefore, a mass production method through fermentation is being sought.

테이코플라닌은 야생균주인 액티노플라네스 테이코마이세티커스 ATCC 31121을 배양하여 배지로부터 회수한 생성물을 정제하여 제조되어 왔으나 14∼20 mg/ℓ의 낮은 생산수율(미국특허 제4,927,754호) 때문에 산업적으로 대량생산 공정에 적용하기에는 부적합하였다.Teicoplanin has been produced by culturing the wild strain Actinoplanes teicomyceticus ATCC 31121 to purify the product recovered from the medium, but low production yield of 14-20 mg / L (US Patent No. 4,927,754) It was therefore unsuitable for industrial application in mass production.

따라서, 산업적으로 이용할 수 있는 고수율의 테이코플라닌 생산균주의 개발이 절실히 요구되고 있는 바, 이에 본 발명자들은 야생균주 액티노플라네스 테이코마이세티커스 ATCC 31121에 비해 테이코플라닌 생산능이 약 25배 이상으로 크게 향상된 변이주 액티노플라네스 테이코마이세티커스 MSL 2211 (기탁번호 : KCTC 8926P)를 개발하여 특허출원한 바 있다.(출원번호: 제10-1999-0013665호, 발명의 명칭: 변이주 액티노플라네스 테이코마이세티커스 MSL 2211 및 그로부터 생산되는 테이코플라닌). 이 선행기술은 수율이 크게 향상되었다는 점에서 고무적이나 증가된 수요를 감안한다면 비교적 낮은 원가로의 원료 공급이 필요하며 보다더 개량된 생산방법이 요구된다.Therefore, the development of industrially available high yield of teicoplanin production strains are urgently required. Therefore, the present inventors compared the wild strain Actinoplanes teicomyceticus ATCC 31121 with the ability to produce teicoplanin. Variant strain Actinoplanes Teicomyceticus MSL 2211 (Accession No .: KCTC 8926P) has been developed and patented (more than 10-1999-0013665). : Variant strain Actinoplanes teicomyceticus MSL 2211 and teicoplanin produced therefrom). This prior art is encouraging in that yields have been greatly improved, but considering the increased demand, it is necessary to supply raw materials at relatively low costs and require more improved production methods.

따라서, 본 발명의 목적은 액티노플라네스 테이코마이세티커스 MSL 2211 (기탁번호 : KCTC 8926P) 의 형질을 개량함으로써 테이코플라닌 고생산 특성을 갖는 신규 변이주를 제공하고, 이를 응용하여 테이코플라닌의 대량 생산 방법을 제공하는데 있다.Accordingly, it is an object of the present invention to provide novel mutants having high teicoplanin production characteristics by improving the trait of Actinoplanes teicomyceticus MSL 2211 (Accession No .: KCTC 8926P), and applying this to teico It is to provide a mass production method of planin.

도 1은 종래 균주 ATCC 31121의 포자를 보여주는 전자현미경 사진이다.1 is an electron micrograph showing the spores of the conventional strain ATCC 31121.

도 2는 종래 균주 MSL 2211 (KCTC 8926P)의 포자를 보여주는 전자현미경 사진이다.2 is an electron micrograph showing the spores of the conventional strain MSL 2211 (KCTC 8926P).

도 3은 본 발명의 변이주 MSL 1510 (KCTC 10200BP)의 포자를 보여주는 전자현미경 사진이다.Figure 3 is an electron micrograph showing the spores of the mutant strain MSL 1510 (KCTC 10200BP) of the present invention.

도 4는 본 발명의 변이주 액티노플라네스 테이코마이세티커스 MSL 1510 균주에서 생산된 시료의 HPLC 결과를 나타내는 그래프이다.Figure 4 is a graph showing the HPLC results of the sample produced in the mutant strain Actinoplanes Teicomyceticus MSL 1510 of the present invention.

본 발명은 테이코플라닌 고생산능을 가진 액티노플라네스 테이코마이세티커스(Actinoplanes teichomyceticus)의 신규 변이주 및 이를 이용한 테이코플라닌의 생산 방법에 관한 것이다.The present invention relates to a novel mutant strain of Actinoplanes teichomyceticus having high teicoplanin production capacity and a method for producing teicoplanin using the same.

본 발명에 따른 신규 변이주 액티노플라네스 테이코마이세티커스 MSL 1510은 야생균주인 액티노플라네스 테이코마이세티커스 ATCC 31121의 변이주인 액티노플라네스 테이코마이세티커스 MSL 2211 (기탁번호 : KCTC 8926P)를 모균주로 하여 인위적 돌연변이법과 단일 포자 분리법을 이용하여 제조된 고농도의 테이코플라닌 생산 변이주이다.The novel mutant Actinoplanes teicomyceticus MSL 1510 according to the present invention is a variant of Actinoplanes teicomyceticus MSL 2211 which is a variant of the wild strain Actinoplanes teicomyceticus ATCC 31121 (Accession No .: KCTC 8926P) as a parent strain is a high concentration of teicoplanin production mutant prepared by artificial mutagenesis and single spore separation method.

이하, 본 발명을 구체적으로 설명한다.Hereinafter, the present invention will be described in detail.

본 발명에서 이용될 수 있는 신규 변이주의 돌연변이 및 선별방법을 요약하면, 돌연변이 유도방법은 물리적 및 화학적 돌연변이 유도처리가 있는 바, 이들 방법 중 물리적 돌연변이 유도방법으로 자외선 및 감마선을 사용할 수 있고, 화학적 유도방법으로는 다양한 변이유도제를 사용할 수 있으며, 세포내 DNA에 직접 작용하는 알킬화제를 사용가능하며, 니트로소구아니딘 (NTG)을 사용할 수 있다.In summary, the mutation and screening methods of novel mutant strains that can be used in the present invention, the mutation induction method has a physical and chemical mutation induction treatment, among these methods can be used to induce ultraviolet light and gamma rays, and chemical induction As a method, various mutagenesis agents can be used, an alkylating agent that directly acts on intracellular DNA can be used, and nitrosoguanidine (NTG) can be used.

변이주의 선별은 상술한 돌연변이 유도처리에 의해 제조된 여러 변이주들을 항균작용을 검사하는 한천확산법 (참조문헌: J. Antibiotics, 615∼620, 1984)을 통해 선별하고, 탄소원, 질소원 및 무기염을 원료로 한 액체 배지에서 배양하여, 배양액을 고압 액체 크로마토그래피(HPLC)를 통해 역가를 확인한 다음, 모균주보다 역가가 크게 향상된 변이주를 분리하는 방법을 반복 실험하여, 테이코플라닌의 생산능이 야생균주보다 약 50배 이상 향상된 변이주를 선별한다.Selection of the mutant strains was carried out by agar diffusion method (J. Antibiotics, 615-620, 1984) to examine the antimicrobial activity of the various strains prepared by the above-described mutagenesis-induced treatment, and the carbon source, nitrogen source and inorganic salt as raw materials Cultured in a liquid medium, and the culture medium was identified by high pressure liquid chromatography (HPLC), and then repeated experiments were performed to isolate mutant strains with significantly higher titers than the parent strain. Variant strains that are about 50 times more advanced are selected.

최종 선별된 가장 우수한 변이주 액티노플라네스 테이코마이세티커스 MSL 1510이라 명명하고, 선행균주와 균학적 성질을 조사, 비교한 후 신균주임을 확인하고, 이 신균주를 한국생명공학연구원(KRIBB)에 2002. 3. 14.자 기탁하여 기탁번호 KCTC 10200BP를 부여받았다.The best variant strain selected was Actinoplanes Teicomyceticus MSL 1510. After checking and comparing the preceding strain with mycological properties, it was confirmed that it was a new strain and the new strain was named Korea Biotechnology Research Institute (KRIBB). Was deposited on March 14, 2002 and was given accession number KCTC 10200BP.

나아가 이 신균주 KCTC 10200BP를 종균배양배지에 접종하여 종배양하고, 얻어진 종배양액을 본배양배지에 접종하여 25-35 ℃, 100∼300 rpm에서 3∼7일간 배양하여 테이코플라닌을 대량생산하는 것이다.Further, the new strain KCTC 10200BP was inoculated in the seed culture medium to incubate, and the obtained seed culture solution was inoculated in the main culture medium and incubated for 3-7 days at 25-35 ° C. and 100 to 300 rpm to mass produce teicoplanin. It is.

이하, 본 발명의 실시예를 들어 상세히 설명하고자 한다.Hereinafter, an embodiment of the present invention will be described in detail.

실시예1 : 신규 변이주 액티노플라네스 테이코마이세티커스 MSL 1510의 제조.Example 1 Preparation of a New Mutant Actinoplanes Teicomyceticus MSL 1510.

테이코플라닌 고생산 변이주를 제조하기 위하여 모균주인 액티노플라네스 테이코마이세티커스 KCTC 8926P를 벤넷(Bennett's) 한천배지에 도말하고, 28℃에서 7일동안 배양한 다음, 유리섬유(제품명:GF/A, 제조회사:Whatman)로 여과하였다. 이어, 상기 여과과정에 의해 수집된 포자를 50mM 트리스-말레이트 완충용액 (pH 8.0)으로 희석하여 106-108포자수/㎖가 되도록 포자 현탁액을 조제하였다.In order to prepare high yielding teicoplanin strains, the parent strain Actinoplanes teicomyceticus KCTC 8926P was plated in Bennett's agar medium, incubated at 28 ° C. for 7 days, and then glass fiber ( : GF / A, manufacturer: Whatman). The spores collected by the filtration process were then diluted with 50 mM Tris-maleate buffer (pH 8.0) to prepare a spore suspension to 10 6 -10 8 spore count / ml.

이 포자용액에 97 %이상의 사멸율이 되도록 0.2 Mrad (106radiation, 1 rad = 100 erg/g, 0.01 joules/kg)의 감마선을 조사하였으며, 형성된 콜로니에 병원성 미생물인 바실러스 서브틸리스(Bacillus subtilis)가 포함된 한천배지를 도포한 다음, 37℃에서 18시간 배양하여 대조균주인 액티노플라네스 테이코마이세티커스 KCTC 8926P 보다 투명환이 크게 나타나는 콜로니를 선발하였다. 한편, 선별된 변이주를 액티노플라네스 테이코마이세티커스 MSL이라 명명하였고, 콜로니에 따라 번호를 부여하였다.This spore solution was irradiated with gamma rays of 0.2 Mrad (10 6 radiation, 1 rad = 100 erg / g, 0.01 joules / kg) to have a mortality of more than 97%. Bacillus subtilis , a pathogenic microorganism, was formed in the colonies. ) Was applied to agar medium, and then incubated at 37 ° C. for 18 hours to select colonies showing clearer rings than the control strain Actinoplanes teicomyceticus KCTC 8926P. On the other hand, the selected mutants were named Actinoplanes Teicomyceticus MSL and numbered according to colonies.

이와같이 분리한 변이주들을 벤넷 액체배양배지 (글루코스, 효모즙, 육즙, 펩톤)에서 28℃에서 일주일동안 배양한 다음, 배양액의 색상변화를 관찰하였으며,그 결과는 다음 표 1과 같다.Thus isolated mutants were cultured for one week at 28 ℃ in Bennett liquid culture medium (glucose, yeast juice, gravy, peptone), and observed the color change of the culture, the results are shown in Table 1 below.

상기 표 1의 배양액 색상을 통해 확인할 수 있듯이, 변이주 중 MSL 1510 배양액의 색상이 특히 주황색으로 상이함을 알 수 있었다.As can be seen from the culture color of Table 1, it was found that the color of the MSL 1510 culture in the mutant strain is particularly different orange.

이어서, 본 발명의 테이코플라닌 고생산 변이주를 최종적으로 선별하기 위하여 일차적으로 선발된 변이주들을 모균주와 같이 표 2의 종균배양액에서 25∼35℃로 2일간 배양한 후 100㎖의 본배양액에 5∼10% (v/v) 접종한 뒤 25∼35℃에서 일주일간 배양한 후 관용의 테이코플라닌 정제방법을 이용하여 부분 정제한 후 관용의 HPLC 정량분석을 통하여 테이코플라닌 농도가 가장 높은 변이주를 얻은 다음, 단일 포자 분리법으로 생산성이 가장 높고 안정된 변이주가 액티노플라네스 테이코마이세티커스 MSL 1510 균주임을 확인하였다. (표 3 참조)Subsequently, in order to finally select the teicoplanin high-producing mutants of the present invention, the first selected mutants were incubated at 25-35 ° C. for 2 days in the seed cultures of Table 2 together with the parent strain, and then in 100 ml of the main culture. After inoculating 5 to 10% (v / v) and incubating for one week at 25 to 35 ° C., partial purification was performed using conventional teicoplanin purification method, followed by conventional HPLC quantitative analysis. After obtaining the highest mutants, the single-spore separation method confirmed that the most productive and stable mutants were Actinoplanes teicomyceticus MSL 1510 strain. (See Table 3)

선별한 변이주인 액티노플라네스 테이코마이세티커스 MSL 1510 균주의 형태학적 특성은 모균주와 거의 비슷하여 타원형 형태의 포자낭을 형성하며 표면은 주름형이었다. (표 4 참조)The morphological characteristics of the selected strain, Actinoplanes teicomyceticus MSL 1510, were almost similar to that of the parent strain, forming an ellipsoidal spore sac and the surface was wrinkled. (See Table 4)

각각의 포자 전자현미경 사진을 도 1, 도 2 및 도 3으로 첨부하였다.Each spore electron micrograph was attached to FIGS. 1, 2 and 3.

배양학적 특성은 조사 대상의 각종 배지에 액티노플라네스 테이코마이세티커스 MSL 1510을 접종하여, 28℃에서 14일 동안 배양한 후 그 생육정도를 관찰하였다. 본 균주의 생육정도는 효모·맥아한천, 오트밀한천, 무기염류·전분한천배지, 글리세롤·아스파라긴한천배지 및 타이로신 한천배지에서 비교적 양호한 생육정도를 나타냈으며 배양액이 주황색을 나타냈다.The culture characteristics were inoculated with Actinoplanes Teicomyceticus MSL 1510 in various media to be investigated, and after culturing for 14 days at 28 ℃ was observed the growth. The growth rate of this strain was relatively good in yeast, malt agar, oatmeal agar, inorganic salt, starch agar medium, glycerol, asparagine agar medium and tyrosine agar medium, and the culture medium was orange.

생리학적 특성으로 최적 pH는 6.5∼7.5 이고, 온도 25∼35℃에서 성장하는 균주이다.The optimum pH is 6.5-7.5, and the strain grows at a temperature of 25-35 ° C.

본 발명에서 분리한 변이주의 생리학적 특성 및 탄소원이용성은 표 5와 같다.Physiological characteristics and carbon source availability of the modified strain isolated in the present invention are shown in Table 5.

상기 표 5 에서 확인할 수 있듯이, 본 발명 액티노플라네스 테이코마이세티커스 MSL 1510은 탄소원이용성 및 배양학적 특성이 모균주인 액티노플라네스 테이코마이세티커스 MSL 2211 (기탁번호 : KCTC 8926P)와는 명확하게 다름을 알 수 있다.As can be seen in Table 5, the Actinoplanes Teicomyceticus MSL 1510 of the present invention is the actinoplanes Teicomyceticus MSL 2211 which is a parent strain of carbon source availability and culture characteristics (Accession Number: KCTC 8926P) It is clearly different from.

상기 과정에 의해 최종 선별된 테이코플라닌 생산력이 가장 우수한 변이주를액티노플라네스 테이코마이세티커스 MSL 1510이라 명명하고, 이를 대한민국 기탁기관이며, 부다페스트조약에 의한 국제기탁기관인 한국생명공학연구원내 유전자원센터에 2002년 3월 14일자로 기탁하였다(수탁번호 KCTC 10200BP).The most highly mutated strain selected by the above process was named Actinoplanes Teicomyceticus MSL 1510, which is a Korean depository institution and an international depository institution under the Budapest Treaty. It was deposited with the Genetic Resource Center on March 14, 2002 (Accession No. KCTC 10200BP).

실시예 2 : 테이코플라닌의 제조.Example 2 Preparation of Teicoplanin.

사용균주 : 변이주 액티노플라네스 테이코마이세티커스 MSL 1510Strains used: Mutant strain Actinoplanes teicomyceticus MSL 1510

배양방법 : 상기 표 2의 종균배양배지액 100㎖ (pH 6.5∼7.5)를 제조하고, 벤넷 한천배지로부터 배양액에 본 발명 액티노플라네스 테이코마이세티커스 MSL 1510을 접종하였다. 이어, 상기 배양액을 25∼35℃에서 100∼300 rpm으로 12∼48시간 동안 종균 배양한 다음 배양액을 상기 표 2의 본 배양배지액 (pH 6.5∼7.5) 100㎖에 5∼10(v/v)%로 접종한 후 25∼35℃, 100∼300 rpm에서 3∼7일간 배양하였다. 이 배양액에 50∼80% 메탄올을 가하여 혼합하고 흔들어 준 다음 원심분리한 후 HPLC로 분석하여 정량하였다. 테이코플라닌 분석은 YMC-Pack ODS-A 컬럼(4.6 ×250mm)과 휴렛펙커드(Hewlett Packard)사의 고압 액체 크로마토그래피 기기(Model: LC SeriesⅡ 1090)를 사용하였고, 표준물질로 주사용 테이코플라닌(타고시드®)을 사용하였다.Cultivation method: 100 ml (pH 6.5-7.5) of the spawn culture medium of Table 2 was prepared, and the culture solution of the present invention was inoculated with Actinoplanes teicomyceticus MSL 1510 from the Bennet agar medium. Subsequently, the culture medium was spawned for 12 to 48 hours at 25 to 35 ° C. at 100 to 300 rpm, and then the culture medium was 5 to 10 (v / v) in 100 ml of the culture medium (pH 6.5 to 7.5) shown in Table 2 above. After inoculation at)%, the cells were incubated at 25-35 ° C. and 100-300 rpm for 3-7 days. 50-80% methanol was added to the culture solution, mixed, shaken, centrifuged, and analyzed by HPLC. Teicoplanin analysis was performed using a YMC-Pack ODS-A column (4.6 × 250 mm) and Hewlett Packard's high pressure liquid chromatography instrument (Model: LC Series II 1090). Planin (Tagosides ® ) was used.

생산된 시료의 HPLC 결과는 도 4와 같으며, 추가로 상기와 동일한 방법으로 수행하되 균주만을 액티노플라네스 테이코마이세티커스 ATCC 31121 또는 액티노플라네스 테이코마이세티커스 MSL 2211 (KCTC 8926P)으로 바꾸어 제조한후 수율비교를 하여 표 6에 정리하였다.The HPLC results of the produced samples are as shown in FIG. 4, and additionally, the strains were performed in the same manner as above, but only strains of Actinoplanes teicomyceticus ATCC 31121 or Actinoplanes teicomyceticus MSL 2211 (KCTC 8926P). ), And the yield comparison was made in Table 6 below.

상기 표 6에 나타난 바와 같이, 본 발명 변이주의 배양액 중 테이코플라닌 농도는 1000∼1500 mg/ℓ로서 모균주에 비해 2배이상, 야생균주에 비해서는 50배 이상으로 높게 생성되었다.As shown in Table 6, the teicoplanin concentration in the culture strain of the present invention was 1000 to 1500 mg / L, two times higher than the parent strain, 50 times higher than the wild strain was produced.

따라서, 본 발명에 따라 테이코플라닌 생산능을 향상시킨 미생물을 제공하여 고부가가치의 테이코플라닌을 산업적으로 대량생산할 수 있는 효과가 있다.Therefore, according to the present invention, by providing a microorganism having improved teicoplanin production capacity, there is an effect of industrially mass-producing high value-added teicoplanin.

Claims (8)

테이코플라닌의 고생산을 특징으로 하는 미생물 액티노플라네스 테이코마이세티커스(Actinoplanes teichomyceticus) MSL 1510 (한국생명공학연구원내 유전자원센터 수탁번호 KCTC 10200BP). Actinoplanes teichomyceticus MSL 1510 (KCTC 10200BP, Korea Institute of Bioscience and Biotechnology) characterized by high production of teicoplanin. 모균주 액티노플라네스 테이코마이세티커스 MSL 2211을 감마선을 사용하여 돌연변이시켜 테이코플라닌을 과량 생산하는 균주를 선별함을 특징으로 하는 테이코플라닌 생성 변이주 액티노플라네스 테이코마이세티커스 MSL 1510 (수탁번호 KCTC 10200BP)의 분리방법.Teicoplanin-producing variant strain Actinoplanes teicomyceti, characterized by mutating the parent strain Actinoplanes teicomyceticus MSL 2211 using gamma rays to select strains that overproduce teicoplanin. Isolation of Curse MSL 1510 (Accession No. KCTC 10200BP). 제2항에 있어서, 감마선의 조사량을 0.2 Mrad (106radiation, 1 rad = 100 erg/g)하여 돌연변이시켜 테이코플라닌을 과량 생산하는 균주를 선별함을 특징으로 하는 테이코플라닌 생성 변이주 액티노플라네스 테이코마이세티커스 MSL 1510(기탁번호 KCTC 10200BP)의 분리방법.3. The teicoplanin producing mutant strain according to claim 2, wherein the strain that overproduces teicoplanin is selected by mutating by irradiating gamma ray with 0.2 Mrad (10 6 radiation, 1 rad = 100 erg / g). Separation method of Actinoplanes teicomyceticus MSL 1510 (Accession No. KCTC 10200BP). 제1항의 테이코플라닌의 고생산을 특징으로 하는 미생물 액티노플라네스 테이코마이세티커스 MSL 1510(한국생명공학연구원 내 유전자원센터 수탁번호 KCTC 10200BP)을 이용하여 테이코플라닌을 생산하는 방법.Producing teicoplanin using microbial Actinoplanes teicomyceticus MSL 1510 (KCTC 10200BP of Korea Research Institute of Bioscience and Biotechnology) characterized by high production of teicoplanin of claim 1 Way. 제4항에 있어서, 액티노플라네스 테이코마이세티커스 MSL 1510(기탁번호 KCTC 10200BP)를 종배양 시킨 후, 포도당 3.0%, 효모추출물 1.0% 및 무기염류를 함유하는 생산배지에서 성장시켜 테이코플라닌을 발효생성 제조하는 방법.The method according to claim 4, wherein after culturing Actinoplanes teicomyceticus MSL 1510 (Accession No. KCTC 10200BP), it is grown in a production medium containing 3.0% glucose, 1.0% yeast extract and inorganic salts. Fermentation production method of the flan. 제5항에 있어서, 상기 무기염류는 황산마그네슘 0.05%, 염화나트륨 0.01% 및 염화칼슘 0.01%를 함유하는 것을 특징으로 하는 테이코플라닌 제조방법.6. The method according to claim 5, wherein the inorganic salts contain 0.05% magnesium sulfate, 0.01% sodium chloride and 0.01% calcium chloride. 제5항에 있어서, 액티노플라네스 테이코마이세티커스 MSL 1510의 종균배양액을 5~10%(v/v)접종하여 제조하는 것을 특징으로 하는 테이코플라닌 제조방법.6. The method for preparing teicoplanin according to claim 5, wherein the seed culture medium of Actinoplanes teicomyceticus MSL 1510 is prepared by inoculation with 5-10% (v / v). 제7항에 있어서, 상기 배양액은 제5항의 배양액으로 구성된 것임을 특징으로 하는 테이코플라닌 제조방법.The method of claim 7, wherein the culture solution is teicoplanin production method characterized in that consisting of the culture solution of claim 5.
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