固定化全细胞法制备生物柴油

发布时间：2018-05-26 12:22

本文选题：生物柴油 + 脂肪酶　；参考：《内蒙古科技大学》2011年硕士论文

【摘要】：随着石油等传统石化能源日益枯竭，以及产生的环境污染问题日益严重，，迫使人们开始寻找清洁、可持续利用的替代能源，生物柴油作为环境友好的可再生的清洁能源备受关注。生物柴油是指由动植物油与短链醇（甲醇或乙醇）进行酯交换反应所制备的脂肪酸甲酯或脂肪酸乙酯。目前采用的化学合成法具有工艺复杂、能耗高、产品色泽深以及污染环境等缺点，而用脂肪酶法催化合成生物柴油具有条件温和、醇用量小、产品易于收集、无污染物排放等优点，可以解决化学法存在的问题，是生物柴油工业重要发展方向。在本研究中，筛选了10株脂肪酶产生菌并对重点对其中酶活力较高的1株脂肪酶产生菌(Y_1)进行催化生产生物柴油的研究。 1.对菌株Y_1进行最佳培养条件确定等研究：菌株Y_1最佳发酵培养基组成为(g/L)：蔗糖5，蛋白胨15，(NH_4)_2SO_41，K_2HPO_41.5， MgSO_4·7H_2O2，橄榄油15，以吐温80为诱导剂。在最佳发酵培养基基础上，30℃、170r/min摇床培养36h，最大产酶活力达73.3U/mL，比初始菌株的酶活提高了4.88倍。 2.对菌株Y_1的甲醇耐受性进行了驯化，4%的甲醇利于产酶。 3.菌株Y_1全细胞的固定化：将菌株Y_1发酵液得到的沉淀进行冷冻干燥，采用聚氨酯泡沫法进行了固定化。 4.固定化全细胞法制备生物柴油：转酯化体系：9.65g橄榄油+0.35g甲醇（分三次加入开始加1.0g，24h后加1.5g，48h后加1.0g）+1.5g有机溶剂(正己烷)+1.2g酶。薄层层析分析转酯化产物。本实验对菌种Y_1产酶条件进行了优化和甲醇耐受性的驯化，并进行了全细胞的固定化研究，通过其催化转酯化反应成功合成了脂肪酸甲酯。
[Abstract]:With the depletion of traditional fossil energy sources, such as petroleum, and the increasing environmental pollution, people are forced to look for clean and sustainable alternative energy sources. Biodiesel, as an environmentally friendly and renewable clean energy source, has attracted much attention. Biodiesel is a kind of fatty acid methyl ester or fatty acid ethyl ester prepared by transesterification of animal and vegetable oil with short chain alcohol (methanol or ethanol). The chemical synthesis method used at present has the disadvantages of complex process, high energy consumption, deep product color and pollution of environment, etc. The lipase catalytic synthesis of biodiesel has mild conditions, small amount of alcohol, and easy to collect. The advantages of no pollutant emission can solve the problems existing in chemical method and are the important development direction of biodiesel industry. In this study, 10 strains of lipase producing bacteria were screened and the catalytic production of biodiesel was carried out. 1. The optimum culture conditions of strain Y1 were determined. The optimum fermentation medium was composed of sucrose (5), peptone (15), NH4 (2SO4), SHP (2HPO41.5), MgSO_4 (7H _ 2O _ 2), olive oil (15), and Twain 80 was used as inducer. On the basis of the optimum fermentation medium, the maximum enzyme producing activity was 73.3 U / mL, 4.88 times higher than that of the original strain. 2. The methanol tolerance of strain Y _ 1 was acclimated to 4% methanol for enzyme production. 3. Immobilization of the whole cell of strain Y-1: the precipitation obtained from the fermentation liquid of strain Y-1 was freeze-dried and immobilized by polyurethane foam method. 4. Preparation of biodiesel by immobilized whole cell method: transesterification system: 1. 65 g olive oil 0.35 g methanol (1. 5 g / h) 1.5 g organic solvent (n-hexane) 1. 2g enzyme. The transesterification products were analyzed by TLC. In this experiment, we optimized the enzyme production conditions and acclimated the methanol tolerance of YStu1 strain, and studied the immobilization of the whole cell. The fatty acid methyl ester was successfully synthesized by its catalytic transesterification reaction.
【学位授予单位】：内蒙古科技大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：TE667

【参考文献】