一株产1-脱氧野尻霉素(DNJ)枯草芽孢杆菌黑色变种的诱变育种研究

Mutation Breeding of a Isolated Bacillus subtilis var. niger for Producing the 1-Deoxynojirimycin

1-脱氧野尻霉素(1-Deoxynojirimycin,DNJ)是一种哌啶生物碱,是α-葡萄糖苷酶的强抑制剂,具有降低餐后血糖、抗肿瘤转移和抗病毒等作用.目前工业生产的DNJ主要是从桑叶中提取的,价格昂贵.为了获得DNJ的稳定高产菌株,本文通过对产DNJ枯草芽孢杆菌黑色变种(Bacillus subtilis var.niger)分别采用紫外线诱变、亚硝基胍诱变和60 Coγ射线诱变方法,得到发酵培养液DNJ表达量达20.7mg/L的菌株,其表达量较初始菌株产量提高了27.7%,并对细菌的DNJ合成代谢相关基因进行了克隆和测序,菌株命名为B-231.

1-Deoxynojirimycin（DNJ）is a kind of alkaloid,known to improve diabetic conditions by inhibiting the activity ofα-glucosidase.It also plays a role in anti-tumor metastasis and antiviral activity.DNJ has been mainly isolated from the root and leaf of the mulberry tree（Morus alba）at present.However,it leads high cost.To obtain stable and high yield DNJ producing strain,in the current study Bacillus subtilis var.niger which can produce DNJ was treated with UV mutagenesis,nitrosoguanidine and 60 Coγ,a strain was isolated and named as B-231 finally.It produced 20.7mg/L DNJ in the fermentation broth,and the content of DNJ increased by 27.7%compared with the initial strain.More research was done on the new strain about its character in molecular biology.The genes associated with anabolism of DNJ were cloned and sequenced.