为探讨诱导子诱导链霉菌702产农抗702的作用机制。将大肠杆菌诱导子添加链霉菌702中,测定链霉菌702发酵过程中生化指标,异柠檬酸脱氢酶(ICDHc)、乙酰辅酶A羧化酶(ACC)、丙酮酸脱氢酶(PDH)酶活测定以及采用凝胶电泳法分析菌体蛋白质谱带变化。结果显示,与对照相比,添加诱导子促进农抗702提前合成,可溶性氨基氮、p H值上升;发酵末期,农抗702的效价提高了47.18%。诱导子添加浓度为3.1μg/m L时,ICDHc、ACC、PDH酶活及总蛋白浓度最大,分别为1.15,0.17,0.75 U,3.38 mg/m L,比对照分别提高了2.24、2.31、0.22、0.22倍。在蛋白质分子质量为20-80 ku,对照蛋白电泳图谱有12条条带,诱导子添加浓度为1.1和3.1μg/m L时,电泳图谱中有14条条带,且有10条带比对照明显增强。试验表明,诱导子促进了农抗702的合成,提高了ICDHc、ACC、PDH酶活及总蛋白浓度,且蛋白质的种类和含量与对照差异显著,可为揭示诱导子促进微生物产次级代谢物的作用机制提供一定的参考。
In order to probe the mechanism of elicitors in inducing Streptomyces 702 to produce antifungalmycin,Escherichia coli elicitor was added into streptomyces 702,biochemical parameters,and the activity of ICDHc,acetyl coenzyme( ACC),pyruvate dehydrogenase( PDH) were determined,and the changes in bacterial protein bands were analyzed with gel electrophoresis. The results showed that the addition of Escherichia coli elicitor advanced the synthesis of antifungalmycin 702 compared with the control,while the soluble amino nitrogen,p H value increased,the titer of antifungalmycin 702 was 47.18% higher than that of the control at the end stage of fermentation.When the concentration of the elicitor 3.1 μg / m L,the activity of ICDHc,ACC,PDH and total protein concentration was at the maximum,amounting to 1. 15,0. 17,0. 75 U,3. 38 mg / m L respectively,2.24,2.31,0.22,0.22 times higher than that of the control. When the protein molecular weight was 20-80 ku there were 12 bands in the control protein electrophoresis map,and the concentrations elicitor were 1. 1 and3.1 μg / m L,there were 14 bands,and 10 bands significantly orhanced with comparing to the control. Tests showed that elicitor promoted the asynthesis of antifungalmycin 702,improved ICDHc,ACC,PDH activity,and the types and content of protein were significant different from those of the control.This study provides a theoretical basis for revealing the mechanism elicitor in promoting production of secondary metabolites produced by microbes.