响应面法优化机械辅助提取放线菌肽聚糖

摘要

在单因素实验的基础上,通过响应面分析法对机械球磨辅助提取放线菌发酵菌泥肽聚糖的提取工艺进行优化.实验结果表明:在NaOH质量分数为6％、球料比为15、转速为300 r/min的条件下作用5 min能够有效去除细胞壁表层磷壁酸,并维持其骨架相对完整.与传统方法相比,机械研磨具有时间短、效率高和经济环保的特点.混合物经过十二烷基磺酸钠(SDS)沸水浴30 min和37℃胰蛋白酶处理6h后得到类白色提取物.经溶菌酶酶解实验和红外光谱分析,初步证明该提取物主要成分为肽聚糖.%Based on the single factor tests,the response surface methodology(RSM) was used for optimizing mechanochemical extraction processing of peptidoglycan from Actinoplanes sp.SE50/ 110.The results showed that the optimum millingcondition was as follows:NaOH(6％),ball to power weight ratio(15),rotating speed(300 r/min).After treated for 5 minutes under the above condition,most of the lipoteichoic acid was removed to release large amount of P with relatively intact cell wall skeleton.Compared to conventional method,the mechanochemical extraction wasmore efficient,economic and environmentally friendly.After the boiling bath with 8 g/dL SDS for 30 min,the residues were incubated in PBS(50 mmol/L,pH 6.8)containing 2 000 U trypsin at 37 ℃ for 6 h to afford off-white solid.The main content of the extract was preliminary confirmed to be peptidoglycan by lysozyme hydrolysis and FTTR analysis.