共轭亚油酸论文产共轭亚油酸丙酸菌的筛选及发酵条件研究.doc

共轭亚油酸论文：产共轭亚油酸丙酸菌的筛选及发酵条件研究 【中文摘要】亚油酸(Linoleic Acid; LA)是人体必需的三大不饱和脂肪酸之一,作为亚油酸异构体的共轭亚油酸(Conjugated Linoleic Acid; CLA)因其具有多种重要生理功能而备受关注。目前CLA的生产主要是采用化学法,而利用生物转化法生产CLA也日渐成为研究的热点。本论文通过建立简单的CLA含量测定方法,从食品中分离筛选出产CLA量较高的丙酸菌菌株,并对筛选所得菌株的培养基和发酵转化条件进行优化以提高其产量,本论文的主要研究内容和结果如下：1.自奶酪中筛选获得具有产CLA能力的菌株共13株,其中菌株PF-3的相对最好,其产CLA能力为38.41μg·mL-1。利用气相色谱-质谱联用(GC-MS)对发酵产物进行检测,进一步确认菌株PF-3具备产CLA的能力。通过常规鉴定手段对菌株PF-3进行鉴定,鉴定结果显示该菌株为费氏丙酸杆菌(Propionibacterium freudenreichii)。2.对不同的培养基进行了初步优化,确定了MRS培养基作为发酵培养基。通过正交实验和响应面法(RSM)优化研究了丙酸杆菌PF-3发酵生产CLA的培养基成分和发酵条件。得到的最优培养基成分为：蛋白胨(2%)、牛肉膏(1%)、酵母膏(0.5%)、葡萄糖(2.5%)、乙酸钠(0.5%)、柠檬酸铵(0.3%)、磷酸氢二钾(0.2%)、硫酸锰(0.02%)、硫酸镁(0.02%)。最佳发酵条件为：菌体接种量10.2%(v/v),LA添加量0.075%,pH6.75,反应温度31℃, Tween-80作为乳化剂(LA/Tween-80为1：20)的条件下,厌氧振荡培养36h。菌株PF-3在优化后的条件下,其CLA产量为63.58μg-mL-1。 【英文摘要】Linoleic acid(LA) is one of the three essential unsaturated fatty acids, the conjugated linoleic acid(CLA) which is the isomer of linoleic acid, has recently attracted much attention because of its important variety of physiological functions. Currently, the mainly method to producing CLA is by chemical approaches, and the biological transformation method to producing CLA is becoming a research hotspot.In this thesis, we screened Propionibacterium strains with high CLA producing capacity from food, then optimized the fermentation medium and fermentation conditions based on the fast determination method of CLA, the main contents and results of this paper are as follows:1. There are 13 strains which producing CLA were screened from cheese. Strain PF-3, has the relatively highest CLA producing ability, the CLA productivity is up to 38.41μg·mL-1. The fermentation products were detected by Gas Chromatography-Mass Spectrometry(GC-MS), and the strain PF-3 has the ability to produce CLA by futher confirmed. According to the analysis of the cell and colony morphology and physio-biochemical characteristics, the bacterium strain PF-3 was identified as Propio