RESUMO
In this study, we devised a novel method to create heterologous producers of lethal antibiotics against host bacteria. Heterologous producers cannot be created when antibiotics are toxic to host bacteria. To overcome this challenge, we developed a novel method involving construction of a combinatorial library with various promoters and screening based on the production. To realize this, we utilized Combi-OGAB (Combinatorial Ordered Gene Assembly in Bacillus subtilis), which technology can effectively construct diverse combinatorial library and accelerate screening procedures. B. subtilis and Gramicidin S were selected as the host bacterium and the targeted antibiotic, respectively. The screened producer from Combi-OGAB screening cycles achieved >30-fold productivity over the lethal level. These results suggest that our strategy has the potential to maximize the phenotypic resistance of host bacteria to create heterologous lethal antibiotic producers.
RESUMO
4,8-Dimethylnonyl sulfate (1) and 3-methyl-4E-decenyl sulfate (2) were isolated from Daphnia pulex as the Daphnia kairomones that induced morphological defense of a freshwater phytoplankton Scenedesmus gutwinskii var. heterospina (NIES-802). The absolute configuration at C4 of 1 was determined by Ohrui's method applied to alcohol 3. The absolute stereochemistry at C3 of 2 was determined by (1)H-NMR analysis of the (R)-1NMA ester of alcohol 11.
Assuntos
Daphnia/química , Feromônios/isolamento & purificação , Feromônios/farmacologia , Fitoplâncton/efeitos dos fármacos , Ésteres do Ácido Sulfúrico/química , Ésteres do Ácido Sulfúrico/isolamento & purificação , Animais , Água Doce , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Feromônios/química , Ésteres do Ácido Sulfúrico/farmacologiaRESUMO
New aliphatic sulfates and sulfamates were isolated from Daphnia pulex as the Daphnia kairomones that induced morphological defense of a freshwater phytoplankton Scenedesmus gutwinskii var. heterospina (NIES-802). Their structures were determined by spectroscopic and synthetic studies.
Assuntos
Daphnia/química , Feromônios/isolamento & purificação , Feromônios/farmacologia , Fitoplâncton/efeitos dos fármacos , Scenedesmus/efeitos dos fármacos , Ácidos Sulfônicos/isolamento & purificação , Ácidos Sulfônicos/farmacologia , Ésteres do Ácido Sulfúrico/isolamento & purificação , Ésteres do Ácido Sulfúrico/farmacologia , Animais , Água Doce , Estrutura Molecular , Feromônios/química , Ácidos Sulfônicos/química , Ésteres do Ácido Sulfúrico/químicaRESUMO
2,6-Dimethylheptyl sulfate (1) and 6-methyloctyl sulfate (3) were isolated from Daphnia pulex as the Daphnia kairomones that induced morphological defense of a freshwater phytoplankton Scenedesmus gutwinskii var. heterospina (NIES-802). The absolute stereochemistry at C2 of 1 was determined by (1)H-NMR analysis of the (R)-MTPA ester of alcohol 2. The absolute configuration at C6 of 3 was determined by Ohrui's method applied to alcohol 4.