乳酸菌素SLNs.ppt

Production of nisin-loaded solid lipid nanoparticles for sustainedantimicrobial activity 乳酸菌素固体脂质纳米粒的持久抗微生物活性的制备 Introduction Nisin ，a polypeptide of 34 amino acids, exists as two variants (A and Z) Nisin is the only bacteriocin considered as Generally Recognized as Safe (GRAS), and the World Health Organization has approved its use as a food additive (WHO, 1969). Nisin is a natural antimicrobial agent that is used as a preservative in heat processed and low pH foods. 但是其生物学活性经常因为与食物中的某些成分（如蛋白质、脂质和促代谢酶等）相互作用而丧失。 Introduction Nisin-loaded SLNs ：provide protection from the food environment and prolong the biological activity and be used as food preservatives. Advantage：controlled drug release、low cytotoxicity、loading both lipophilic and hydrophilic drugs、large scale production Materials Nisin powder (2.5% (w/w) pure nisin in denatured milk solids,approximately 1,000,000 IU/g)（从变性固体乳中提出的纯乳酸菌素粉） The solid lipids (Cetylpalmitate（十六烷基棕榈酸酯）, Softisan 378, Softisan 154, Imwitor 900 and Witepsol E85) The surfactant Poloxamer 188（泊洛沙姆188） the co-surfactant sodium deoxycholate（脱氧胆酸钠） Lactobacilli MRS medium（乳酸菌MRS培养基） analytical grade chemicals Materials Bacterial strains： Listeria monocytogenes DMST 2871 （单核细胞增生型李氏杆菌） Lactobacillus plantarum TISTR 850（胚芽乳杆菌） Methods and Results 1、Nisin solubility in melted solid lipids 将四种不同浓度的乳酸菌素0.5%， 1.0%，2.0% ， 3.0%分别溶解在80℃5组不同的熔融固体脂质中制成预混液，肉眼观察其溶解度，挑选最合适的固体基质的组成。 结果：除Imwitor 900外，均表现出相对较低的溶解度，故选用Imwitor 900作为唯一的固体基质。 Methods and Results 2、Stability test of unloaded SLN dispersions 分别在匀质压力为500、1000、1500Pa下获得的未包埋乳酸菌素的SLN，在磷酸盐缓冲液中30℃孵育7天及14天，测得其的直径（A)和电势（B） 最终选择匀质压力为1500Pa，表面活性剂为5%（w/w）泊洛沙姆188及0.125%（w/v）的脱氧胆酸盐，可制得在14天的观察期内，粒子直径最小（119±15.1nm）及最低电势（-34.0到-30.2mV）的SLN，这种条件下获得多分散性指数(PDI) 为0.38±0.03。 Methods and Results 3、Preparation of nisin-loaded SLNs 高温高压匀质法 1、将含有10%的Imwitor900固体脂质在高于其熔点5-10℃左右熔化 2、加入同温度的5.0%泊洛沙姆188和0.125%脱氧胆酸钠表面活性剂的水相中 3、进行高速乳匀（Ultra-Turrax T25，8000rpm/min）得到预混乳液 4、将预混乳液在熔点温度下高压乳匀（Em