Colorimetric Method in the Determination of 1-Deoxynojirimycin Isolated from the Mutant Strain of Bacillus subtilis

doi:10.13560/j.cnki.biotech.bull.1985.2016.06.027

Abstract

Abstract: This work is to seek a novel method to detect 1-Deoxynojirimycin(DNJ)in the culture broth of Bacillus subtilis var. 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 of bacteria. 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.

Key words: Bacillus subtilis, 1-deoxynojirimycin, α-glucosidase inhibitor, ultraviolet mutation

LONG Ling, YANG Yan, ZHU Nai-shuo. Colorimetric Method in the Determination of 1-Deoxynojirimycin Isolated from the Mutant Strain of Bacillus subtilis[J]. Biotechnology Bulletin, 2016, 32(6): 187-192.

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