Project description:Streptomyces bingchenggensis is a soil bacterium that produces milbemycins. Milbemycins and their derivatives are valuable biopesticides in the agricultural field. Owing to their advantages such as high efficiency and safety for human and animal,it was urgent to construct high-yield strain to ensure low production cost. To obtain genes closely correlated with milbemycin production, we have compared the whole genome microarray expression profiling of two strains (the parent one strain and high-yielding strain). In Streptomyces bingchenggensis, there are abundant exporters, which are responsible for transporting various substrates. In the result, some drug exporters were chosen to enhance production of milbemycin .
Project description:Streptomyces bingchenggensis is a soil bacterium that produces milbemycins. Milbemycins are commercially insecticidal and acaricidal antibiotics in agriculture, owing to their advantages such as high efficiency and safety for human and animal. To obtain genes valuable for further improvement of titer, we have compared two strains(the parental strain and high-yielding strain)by whole genome microarray expression profiling as a discovery platform. In Streptomyces bingchenggensis, there are abundant transporters, which are responsible for transporting various substrates. In the result, some sugar transporter genes showed significant gene expression.
Project description:<p>We used time-resolved metabolic footprinting, an important technical approach used to monitor changes in extracellular compound concentrations during microbial growth, to study the order of substrate utilization (i.e., substrate preferences) and kinetics of a fast-growing soil isolate, <em>Paraburkholderia</em> sp. strain 1N. The growth of <em>Paraburkholderia</em> sp. 1N was monitored under aerobic conditions in a soil-extracted solubilized organic matter medium, representing a realistic diversity of available substrates and gradient of initial concentrations. We combined multiple analytical approaches to track over 150 compounds in the medium and complemented this with bulk carbon and nitrogen measurements, allowing estimates of carbon use efficiency throughout the growth curve. Targeted methods allowed the quantification of common low-molecular-weight substrates: glucose, 20 amino acids, and 9 organic acids. All targeted compounds were depleted from the medium, and depletion followed a sigmoidal curve where sufficient data were available. Substrates were utilized in at least three distinct temporal clusters as <em>Paraburkholderia</em> sp. 1N produced biomass at a cumulative carbon use efficiency of 0.43. The two substrates with highest initial concentrations, glucose and valine, exhibited longer usage windows, at higher biomass-normalized rates, and later in the growth curve. Contrary to hypotheses based on previous studies, we found no clear relationship between substrate nominal oxidation state of carbon (NOSC) or maximal growth rate and the order of substrate depletion. Under soil solution conditions, the growth of <em>Paraburkholderia</em> sp. 1N induced multiauxic substrate depletion patterns that could not be explained by the traditional paradigm of catabolite repression.</p><p><strong>IMPORTANCE:</strong> Exometabolomic footprinting methods have the capability to provide time-resolved observations of the uptake and release of hundreds of compounds during microbial growth. Of particular interest is microbial phenotyping under environmentally relevant soil conditions, consisting of relatively low concentrations and modeling pulse input events. Here, we show that growth of a bacterial soil isolate, <em>Paraburkholderia</em> sp. 1N, on a dilute soil extract resulted in a multiauxic metabolic response, characterized by discrete temporal clusters of substrate depletion and metabolite production. Our data did not support the hypothesis that compounds with lower energy content are used preferentially, as each cluster contained compounds with a range of nominal oxidation states of carbon. These new findings with <em>Paraburkholderia</em> sp. 1N, which belongs to a metabolically diverse genus, provide insights on ecological strategies employed by aerobic heterotrophs competing for low-molecular-weight substrates in soil solution.</p>