Wang et al Anticancer Exopolysaccharide From B breve Iwo1 1-Ara.2011 10 24 5Ma=11521 6420 246 224 FIGURE 6 Chromatogram of standard monosaccharide(A) and EPS monosaccharide compositions(B)on HPAEC-PAD. From left to right: 1. Rhamnose 2. Arabinose 3. Galactose 4. Glucose 5 Mannose 6. Fructose infant feces. After sequencing its genome, we predicted its 2016; Bengoa et al., 2018). In our study, slimmed milk was chosen EPS cluster and hypothetic synthesis pathway. Based on the for the initial cultivation to avoid sugar in the medium that EPS extraction and high-quality purification, we determined interferes with the EPS extraction. Crude bifido-EPS was followed its chemical composition. Finally, we focused our attention on by the further purification(Ion exchange chromatography and its anticancer activity and provided biochemical evidence to gel filtration chromatography), which yielded high purity bifido- explain it. Thus, this study provided a better understanding of EPS, thus providing good research basis for the biological activity. the function-genome cluster relation and biosynthesis in bifido- Multiple purification induced the poor yield, which was affected EPS, which could facilitate genetic and metabolic engineering by many factors such as EPS sample loading volume, eluent indispensable for production of tailor-made EPS used in food and elution rate. In the course of the research, we observed or drug industry that the low purification efficiency affected our further research Extraction and purification were necessary to examine the Therefore, future studies should focus on the elevation of the biological activity of B breve lwO1's EPS. The yield of crude EPs purification efficiency of EPS isolation. released to culture medium can be influenced by environmental Previous studies have indicated that genes responsible for the and nutritional conditions, such as the medium composition, pH, biosynthesis of EPS are presented in the form of clusters in temperature, oxygen and the growth phase (Leroy and De vuyst, Bifidobacterium genome(Leivers et al, 2011; Fanning et al., 201 FrontiersinMicrobiologywww.frontiersin.org May 2019 Volume 10 Article 1044fmicb-10-01044 May 8, 2019 Time: 14:36 # 8 Wang et al. Anticancer Exopolysaccharide From B. breve lw01 FIGURE 6 | Chromatogram of standard monosaccharide (A) and EPS monosaccharide compositions (B) on HPAEC-PAD. From left to right: 1. Rhamnose 2. Arabinose 3. Galactose 4. Glucose 5. Mannose 6. Fructose. infant feces. After sequencing its genome, we predicted its EPS cluster and hypothetic synthesis pathway. Based on the EPS extraction and high-quality purification, we determined its chemical composition. Finally, we focused our attention on its anticancer activity and provided biochemical evidence to explain it. Thus, this study provided a better understanding of the function-genome cluster relation and biosynthesis in bifido￾EPS, which could facilitate genetic and metabolic engineering indispensable for production of tailor-made EPS used in food or drug industry. Extraction and purification were necessary to examine the biological activity of B. breve lw01’s EPS. The yield of crude EPS released to culture medium can be influenced by environmental and nutritional conditions, such as the medium composition, pH, temperature, oxygen and the growth phase (Leroy and De Vuyst, 2016; Bengoa et al., 2018). In our study, slimmed milk was chosen for the initial cultivation to avoid sugar in the medium that interferes with the EPS extraction. Crude bifido-EPS was followed by the further purification (Ion exchange chromatography and gel filtration chromatography), which yielded high purity bifido￾EPS, thus providing good research basis for the biological activity. Multiple purification induced the poor yield, which was affected by many factors such as EPS sample loading volume, eluent and elution rate. In the course of the research, we observed that the low purification efficiency affected our further research. Therefore, future studies should focus on the elevation of the purification efficiency of EPS isolation. Previous studies have indicated that genes responsible for the biosynthesis of EPS are presented in the form of clusters in Bifidobacterium genome (Leivers et al., 2011; Fanning et al., 2012; Frontiers in Microbiology | www.frontiersin.org 8 May 2019 | Volume 10 | Article 1044