琼脂平板及紫外分光光度法筛选林可霉素降解菌

SCREENING OF LINCOMYCIN-DEGRADING BACTERIA BY AGAR PLATE AND UV SPECTROPHOTOMETRY

由于抗生素的残留,林可霉素生产废渣难以处理及资源化利用,这已成为制药企业的一大难题。因此亟需寻找一种简便而又快捷的方法分离林可霉素降解菌。本研究采用林可霉素作为唯一碳源和能源的固体琼脂平板来筛选林可霉素降解菌,通过加热酸溶解琼脂后采用紫外分光光度法测定琼脂平板中的林可霉素含量来考察降解率。结果表明酸溶解加热的最优工艺组合为每10mL培养基的平板中加入3mL浓度为1.14mol/L的盐酸,于80°C加热70s,在该工艺条件下,在0～250μg/mL的浓度范围内,琼脂平板中林可霉素浓度与吸光值呈良好的线性关系,具有良好的精密度、重现性、稳定性及回收率,表明采用紫外分光法测定琼脂平板中林可霉素含量是合理可行的。采用该方法最终获得1株降解率为(40.63±2)%的降解菌。这为林可霉素降解菌的筛选提供了一种简单、方便、快捷的方法。

Due to the existence of antibiotic residues, waste mycelium of lincomycin was difficult to be treated and used as resources, which had become a major problem in the production enterprise. So it is very necessary to develop a simple and rapid method to isolate and screen lincomycin - degrading bacteria. Solid agar plates utilizing lincomycin as the sole source for carbon and energy were adopted to screen lincomycin-degrading bacteria. The concentration of lincomycin in the agar plate was determined to indicate the degradation capacity using the ultraviolet spectrophotometry analysis, after dissolving the agar by adding with hydrochloric acid and heating. The optimal conditions were found to add 3 mL of 1.14 mol/L hydrochloric acid to 10 mL agar solution, and heat it at 80 ℃ for 70 s. Under these conditions, the linear correlation between the lincnmycin concentration and absorbance was satisfactory in the calibration standards at the range of 0 - 250 μg/mL. The method precision values, accuracy values, reproducibility values, stability values and recovery values of lincomycin in agar solution were adequate. Application of this method to 0409 strain showed the lincomycindegradation rate was of （40.63 ± 2） %. Taken together, we have established a simple, convenient, rapid and valid ultraviolet spectrophotometry method to detect lincomycin in agar plates for screening lineomyein-degrading bacteria.