School of Bioengineering, Qilu University of Technology (Shandong Academy of Sciences), Jinan 250300, PR China.
CAS Key Laboratory of Biobased Materials, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao 266101, PR China.
Int J Biol Macromol. 2019 Dec 1;141:298-306. doi: 10.1016/j.ijbiomac.2019.08.217. Epub 2019 Aug 26.
Extraction polysaccharide from microorganism is a research hotspot. In this work, a new type of water-soluble exopolysaccharides (EPS) was isolated from Bacillus licheniformis. Firstly, response surface methodology (RSM), based on a three-level, three-factor, was used to determine optimum conditions for EPS extraction. And RSM analysis indicated optimum condition was at the temperature of 8 °C for 10.44 h with ethanol at a concentration of 79.22% (v/v), the maximum yield of EPS was 3.07 g/mL. Secondly, EPS were seperated using DEAE-Sepharose Fast Flow column chromatography and acquired two polysaccharide fractions, BL-P1 and BL-P2. BL-P1 had larger molecular weight than BL-P2 from structural analyses, because of higher content of mannose, ribose, glucuronic acid, galactose, arabinose and fructose in BL-P2. Moreover, the characterization of BL-P1 and BL-P2 was investigated with Fourier transform infrared spectroscopy (FT-IR) and nuclear magnetic resonance (NMR) spectroscopy, the results indicated that EPS was mainly composed of→3)-α-d-Galp-(1→, →3,5)-α-l-Araf-(1→, →3)-β-d-Glcp-(1→, β-d-Glcp-(1 → and→4)-β-l-Fucp-(1 → 4)-β-d-Xylp-(1 → 4)-α-l-Rhap (1 → 3) -β-d-Manp-(4 → residues. In vitro antioxidant activity assay, EPS exhibited potent quenching capacities on hydroxyl and 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radicals in a dose-dependent manner. Furthermore, BL-P2 had higher activity than BL-P1 in inhibiting α-amylase and α-glucosidase, which would have potential to be applied in nutraceutical and pharmaceutical industries.
从微生物中提取多糖是一个研究热点。在这项工作中,从地衣芽孢杆菌中分离得到了一种新型水溶性胞外多糖(EPS)。首先,采用基于三水平三因素的响应面法(RSM)确定 EPS 提取的最佳条件。RSM 分析表明,最佳条件为温度 8°C、时间 10.44h、乙醇浓度 79.22%(v/v),EPS 的最大产量为 3.07g/mL。其次,采用 DEAE-Sepharose Fast Flow 柱层析法分离 EPS,得到两个多糖级分 BL-P1 和 BL-P2。结构分析表明,BL-P1 的分子量大于 BL-P2,因为 BL-P2 中甘露糖、核糖、葡萄糖醛酸、半乳糖、阿拉伯糖和果糖的含量较高。此外,通过傅里叶变换红外光谱(FT-IR)和核磁共振(NMR)光谱对 BL-P1 和 BL-P2 的特性进行了研究,结果表明 EPS 主要由→3)-α-d-Galp-(1→、→3,5)-α-l-Araf-(1→、→3)-β-d-Glcp-(1→、β-d-Glcp-(1→和→4)-β-l-Fucp-(1→4)-β-d-Xylp-(1→4)-α-l-Rhap-(1→3)-β-d-Manp-(4→残基组成。体外抗氧化活性测定结果表明,EPS 对羟基和 1,1-二苯基-2-苦基肼(DPPH)自由基具有较强的猝灭能力,且呈剂量依赖性。此外,BL-P2 对α-淀粉酶和α-葡萄糖苷酶的抑制活性高于 BL-P1,具有在营养保健品和制药行业应用的潜力。