You Hyun Ju, Ahn Hyung Jin, Kim Jin Yong, Wu Qian Qian, Ji Geun Eog
Department of Food and Nutrition, Research Institute of Human Ecology, Seoul National University, Seoul 151-742, Republic of Korea.
Research Institute, Bifido Co.,Ltd., Hongchun 250-804, Republic of Korea.
J Microbiol Biotechnol. 2015 Apr;25(4):469-78. doi: 10.4014/jmb.1408.08013.
In spite of the reported probiotic effects, Bifidobacterium bifidum BGN4 (BGN4) showed no β- glucosidase activity and failed to biotransform isoflavone glucosides into the more bioactive aglycones during soy milk fermentation. To develop an isoflavone-biotransforming BGN4, we constructed the recombinant B. bifidum BGN4 strain (B919G) by cloning the structural β- glucosidase gene from B. lactis AD011 (AD011) using the expression vector with the constitutively active promoter 919 from BGN4. As a result, B919G highly expressed β- glucosidase and showed higher β-glucosidase activity and heat stability than the source strain of the β-glucosidase gene, AD011. The biotransformation of daidzin and genistin compounds using the crude enzyme extract from B919G was completed within 4 h, and the bioconversion of daidzin and genistin in soy milk during fermentation with B919G also occurred within 6 h, which was much faster and higher than with AD011. The incorporation of this β-glucosidaseproducing Bifidobacterium strain in soy milk could lead to the production of fermented soy milk with an elevated amount of bioavailable forms of isoflavones as well as to the indigenous probiotic effects of the Bifidobacterium strain.
尽管有报道称双歧双歧杆菌BGN4(BGN4)具有益生菌作用,但在豆浆发酵过程中,它没有表现出β-葡萄糖苷酶活性,也未能将异黄酮葡萄糖苷生物转化为生物活性更高的苷元。为了培育一种能生物转化异黄酮的BGN4,我们通过使用带有来自BGN4的组成型活性启动子919的表达载体,从乳酸双歧杆菌AD011(AD011)中克隆结构β-葡萄糖苷酶基因,构建了重组双歧双歧杆菌BGN4菌株(B919G)。结果,B919G高表达β-葡萄糖苷酶,并且与β-葡萄糖苷酶基因的来源菌株AD011相比,表现出更高的β-葡萄糖苷酶活性和热稳定性。使用来自B919G的粗酶提取物对黄豆苷和染料木苷化合物的生物转化在4小时内完成,并且在B919G发酵豆浆过程中黄豆苷和染料木苷的生物转化也在6小时内发生,这比AD011快得多且效率更高。将这种产生β-葡萄糖苷酶的双歧杆菌菌株添加到豆浆中,可以生产出异黄酮生物可利用形式含量升高的发酵豆浆,以及发挥双歧杆菌菌株的固有益生菌作用。