• 한국미생물·생명공학회지
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• 제23권4호
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• pp.384-389
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• 1995

Streptoalloteichus hindustanus ATCC 31219, a nebramycin complex producer, is similar to Streptomyeces tenebrarius in a viewpoint of resistance to a wide range of aminoglycoside antibiotics. S. tenebrarius has resistance mechanisms of 16s rRNA methylation and aminogycoside modification. However, it is not known whether resistance mechanisms of Stall. hindustanus are the same as in S. tenebrarius. Therefore, we have tried to isolate resistance determinants from Stall. hindustanus. Two different types of aminoglycoside resistance determinants were isolated from Stall. hindustanus and expressed in Streptomyces lividans TK24. The apramycin resistance gene (amr) and the tobramycin resistance gene (tmr) isolated from Stall. hindustanus showed broad resistance spectrum against a dozen of aminoglycoside antibiotics. The complete nucleotide sequences of apramycin resistance gene (amr) were determined. The deduced amino acid sequence of the amr gene of Stall hindustanus ATCC 31219 showed extensive sequence homology to the 16s rRNA methylase gene (kamB) of S. tenebrarius.

• 한국미생물·생명공학회지
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• 제31권4호
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• pp.395-399
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• 2003

본 연구에서는 S. hindustanus ATCC 31218로부터 선별된 변이주를 사용하여 배양조건이 nebramycin 주요 생산물인 factor 2, factor 4, factor 5', kanamycin A 등의 구성비율과 역가에 미치는 영향을 조사하였다. 실험결과, nebramycin의 factor들이 구성비율은 온도 및 배지보다는 배양액량, 교반속도 등에 의해 크게 영향을 받는 것으로 나타났다. 따라서 플라스크와 발효조의 산소전달속도를 각각 측정하여 factor 5' 생산의 최적 산소전달조건을 검토한 결과, $0.50 mMO_2$/min으로 배양시 factor 5'의 역가가 가장 높았으며 구성비율도 70% 이상을 나타내었다. 한편 $0.9 mMO_2$/min 이상으로 배양시는 factor 2의 역가와 구성비율이 현저히 감소한 반면 factor 2의 역가와 구성비율이 급격히 증가하였다. 한편 본 연구에 사용된 변이주 S. hindustanus YHT-0001는 factor 5'의 역가는 16.4배, factor 5'구성비율은 약 11% 향상된 균주로 확인되었다. S. hindustanus 발효시 공급되는 산소량은 단지 hydroxyl기와 아미노기의 존재유무에 따라 구조가 결정되는 factor 5', factor 4, kanamycin A의 hydroxyl기 생성을 위한 산소공여자로 역할뿐 만 아니라 배당체 구조에 차이를 보이는 apramycin 생합성과정에 영향을 크게 준다고 사료되었다.

• 약학회지
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• 제37권1호
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• pp.1-8
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• 1993

A procedure for the high-performance liquid chromatographic determination of Nebramycin factors in fermentation broth of Streptoalloteichus hindustanus ATCC 31218 mutant was investigated using pre-column derivatization and LTV detection. The method is based on pre-column derivatization of Nebramycin factors with 2,4-dinitrofluorobenzene(DNFB) in the presence of Tris (hydroxymethyl)aminoethane. The chromatographic separation of derivatives of Nebramycin factors and unknown impurities is achieved using reversed-phase column (NOVA-PAK $C_{18}$, Waters Co.) and AcCN : H$_{2}$O : AcOH (53.0:46.5:0.5) as a mobile phase. The mixture of these derivatives were separated within 35 minutes and the optimum wavelength($\lambda_{max}$ ) of the UV detector was 353 nm. The linearity of response for derivatives of Nebramycin factors is demonstrated for concentrations up to 500 $\mu\textrm{g}$/ml and the relative standard deviation is less than 0.79%. Detection limit was 1.67 ng for the 10 $\mu\textrm{l}$ sample volume employed.

• Journal of Microbiology and Biotechnology
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• 제8권2호
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• pp.146-151
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• 1998

The aminoglycoside multiple-resistance determinant from Streptoalloteichus hindustanus was cloned into Streptomyces lividans and named nbrB. The 1.2-kb ApaI- BclI fragment encompassing nbrB was located within a 2.6-kb ApaI fragment by successive subcloning experiments. The complete DNA nucleotide sequence of 1.2-kb containing nbrB was determined. The sequence contains an open reading frame that putatively encodes a polypeptide of 281 amino acids with a predicted molecular weight of 30,992. The deduced amino acid sequence of nbrB shows identities of 85.1% to kgmB of S. tenebrarius, 59.6% to sgm of Micromonospora zionensis, and 57.7% to grm of M. rosea. The similarity of nbrB to kgmB suggests that nbrB encodes a 16S rRNA methylase similar to that encoded by kgmB and that both genes might be derived from a common ancestral gene.