为寻找测定枯草芽孢杆菌黑色变种突变株Bacillus subtilis var. niger-231（B-231）的发酵液中1-脱氧野尻霉素（1-Deoxynojirimycin，DNJ）的新方法并提高其产发酵液中DNJ的含量。在有铜离子的存在下，DNJ与二硫化碳反应，用有机溶剂对产物萃取后在440nm进行光度测定，得到DNJ含量。再采用紫外诱变方法处理突变株B-231。绘制了检测DNJ标准品的标准曲线，对测定DNJ的新方法做了精密度、灵敏度、准确度以及产物的稳定性的探讨，设计正交试验优化了反应条件，并将此方法运用到细菌发酵液中DNJ含量的测定中，通过遗传诱变手段处理原始细菌B-231，最终筛选到一株高产DNJ的菌株，命名为B. subtilis var. niger-431（B-431），DNJ含量达87.3mg/L，其产生DNJ的量较原始菌株原突变株提高了4.2倍。首次提出了一种快速测定细菌发酵液中的1-脱氧野尻霉素的新方法，紫外诱变手段使细菌产生DNJ的能力有较大提高。%This work is to seek a novel method to detect 1-Deoxynojirimycin(DNJ)in the culture broth of Bacillus subtilisvar. niger-231 (B-231)and to improve DNJ production. DNJ reacts rapidly with carbon disulfide while Cu2+ existing,and the final product extracted by organic solvent was measured by spectrophotometer at 440 nm,thus DNJ content was determined. Then mutant strain B-231 was induced with ultraviolet. The standard curve for measuring standard DNJ was drawn,and the precision,sensitivity,accuracy and stability of the products while using the novel method of measuring DNJ were studied. Orthogonal test was designed to obtain the optimal reaction condition. The novel method was applied to measure the content of DNJ in the culture broth ofbacteria. A high-yield DNJ strain was screened and named as B. subtilis var. niger- 431(B-431)by mutagenizing the B-231. The content of DNJ by B-431 was up to 87.3 mg/mL,and increased 4.2 times compared to B-231. This study established a novel method to determine 1-deoxynojirimycin in the culture broth of DNJ-producing strain for the first time. The induction by ultraviolet increased the capacity of this bacterium producing DNJ.
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