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Development of the Agar Disk Method for the Rapid Selection of Cephalosporin Producers with Improved Yields

机译:琼脂圆盘法快速提高产量的头孢菌素生产者的开发

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摘要

To screen the abilities of mutant strains of Cephalosporium to produce cephalosporin C, colonies of the organism were grown on the surface of small (4-mm diameter) disks of agar medium. After incubation of the disks for periods of up to 5 days, the antibiotic contents of the disks were assayed by placing them on agar plates of the assay organism and determining the diameters of the inhibition zones. The amount of nitrogen source in the agar disk medium was used to control the amount of antibiotic produced in the disk and, thus, the sensitivity of screening. The relation of agar disk inhibition zone diameters to log shake-flask titers was linear with short incubation times (2 to 3 days) of the disks, but shifted towards a higher order with prolonged incubation (4 to 5 days). The optimum incubation time for the disks was 4 to 5 days, and then a 15% difference in zone diameters was significant with 10 disks per sample. The minimum difference between the shake-flask titers, which could be detected by the agar disk method with 10 disks per sample, was about 30% with 5 days of incubation for the disks. The results suggest that the shake-flask culture underestimated the degree of improvement in strain productivity.
机译:为了筛选头孢菌属突变菌株产生头孢菌素C的能力,在小(4毫米直径)琼脂培养基盘的表面上生长了该菌落。在将盘孵育最多5天之后,通过将盘放置在测定生物的琼脂板上并测定抑制区的直径来测定盘中的抗生素含量。琼脂圆盘培养基中的氮源数量用于控制圆盘中产生的抗生素数量,从而控制筛选的敏感性。琼脂盘抑菌圈直径与对数摇瓶滴度的关系在较短的培养时间(2-3天)下呈线性关系,但在延长的培养时间(4至5天)下向较高位移动。圆盘的最佳孵育时间为4至5天,然后每个样品10个圆盘的区域直径差异为15%。摇瓶滴度之间的最小差异可以通过琼脂圆盘法(每个样品10个圆盘)检测到,孵育5天后大约为30%。结果表明,摇瓶培养低估了菌株生产力的提高程度。

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