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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제25권 제7호
- 발행연도
- 2015.1
- 수록면
- 1,015 - 1,025 (11page)
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The development of diverse polymyxin derivatives is needed to solve the toxicity and resistance problems of polymyxins. However, no platform has generated polymyxin derivatives by genetically engineering a polymyxin synthetase, which is a nonribosomal peptide synthetase. In this study, we present a two-step approach for the construction of engineered polymyxin synthetases by substituting the adenylation (A) domains of polymyxin A synthetase, which is encoded by the pmxABCDE gene cluster of Paenibacillus polymyxa E681.
First, the seventh L-threonine-specific A-domain region in pmxA was substituted with the Lleucine- specific A-domain region obtained from P. polymyxa ATCC21830 to make polymyxin E synthetase, and then the sixth D-leucine-specific A-domain region (A6-D-Leu-domain) was substituted with the D-phenylalanine-specific A-domain region (A6-D-Phe-domain) obtained from P. polymyxa F4 to make polymyxin B synthetase. This step was performed in Escherichia coli on a pmxA-containing fosmid, using the lambda Red recombination system and the sacB gene as a counter-selectable marker. Next, the modified pmxA gene was fused to pmxBCDE on the chromosome of Bacillus subtilis BSK4dA, and the resulting recombinant strains BSK4-PB and BSK4-PE were confirmed to produce polymyxins B and E, respectively. We also succeeded in constructing the B. subtilis BSK4-PP strain, which produces polymyxin P, by singly substituting the A6-D-Leu-domain with the A6-D-Phe-domain. This is the first report in which polymyxin derivatives were generated by genetically engineering polymyxin synthetases. The two recombinant B. subtilis strains will be useful for improving the commercial production of polymyxins B and E, and they will facilitate the generation of novel polymyxin derivatives.
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참고문헌 (24)
참고문헌 신청
Chihara S / 1973 / Enzymatic degradation of colistin isolation and identification of α-N-acyl α, γ-diaminobutyric acid and colistin nonapeptide / Agric. Biol. Chem 37 : 24
Choi S-K / 2009 / Identification of a polymyxin synthetase gene cluster of Paenibacillus polymyxa and heterologous expression of the gene in Bacillus subtilis / J. Bacteriol
Doekel S / 2002 / Heterologous expression of nonribosomal peptide synthetases in B. subtilis:construction o f a b i-functional B. subtilis/E. coli shuttle vector system / FEM
Eppelmann K / 2001 / Engineered biosynthesis of the peptide antibiotic bacitracin in the surrogate host Bacillus subtilis / J. Biol. Chem 276 : 34824 ~ 34831
Evans ME / 1999 / Polymyxin B sulfate and colistin: old antibiotics for emerging multiresistant gram-negative bacteria / Ann. Pharmacother 33 : 960 ~ 967
Choi S-K / 2009 / Identification of a polymyxin synthetase gene cluster of Paenibacillus polymyxa and heterologous expression of the gene in Bacillus subtilis / J. Bacteriol
Doekel S / 2002 / Heterologous expression of nonribosomal peptide synthetases in B. subtilis:construction o f a b i-functional B. subtilis/E. coli shuttle vector system / FEM
Eppelmann K / 2001 / Engineered biosynthesis of the peptide antibiotic bacitracin in the surrogate host Bacillus subtilis / J. Biol. Chem 276 : 34824 ~ 34831
Evans ME / 1999 / Polymyxin B sulfate and colistin: old antibiotics for emerging multiresistant gram-negative bacteria / Ann. Pharmacother 33 : 960 ~ 967
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