【作者】 王瑶；

【导师】 董庆霖； 孙延宁；

【作者基本信息】 河北工业大学 ， 工程硕士（专业学位）， 2019， 硕士

【摘要】 雨生红球藻因主要脂肪酸组份为棕榈酸、油酸和亚油酸等中性脂肪,不饱和双键较少等原因,被认为是生物柴油的理想原料。然而,较高的生产成本限制其大规模工业化生产。本课题希望通过提高雨生红球藻生长速率和脂肪含量来分别提高单位面积产量和降低单位生物量的生产加工成本,从而提高该生产过程的经济性和可持续性。因此,本实验将雨生红球藻与淡紫拟青霉TD16(以下简称TD16)混合培养,以研究真菌对微藻生长和脂类合成的影响,主要研究结果如下:TD16胞外蛋白和多糖都可促进雨生红球藻生物量和脂类含量增加,且TD16胞外蛋白对藻细胞的促进程度大于多糖,同时发现TD16胞外蛋白对金黄色葡萄球菌、枯草芽孢杆菌、大肠杆菌具有抑菌性。将TD16孢子悬浮液接种至含雨生红球藻胞外有机碳的BBM培养基中培养,结果显示TD16的生物量不断增加,同时培养基中胞外有机碳的浓度不断降低。本章确定了雨生红球藻-TD16混合培养的可行性。通过单因素实验,得到最佳培养条件:营养模式、藻菌接种比例(雨生红球藻和TD16的初始浓度分别为1×10~7cells/m L和1.36×10~7cells/m L)、硝酸钠浓度、培养基p H、光照强度和培养温度分别为光合自养、5:1、0.35 g/L、7、70μmol/(m~2·s)和25℃。雨生红球藻-TD16混合培养与雨生红球藻单独培养相比:细胞干重(0.99 g/L)和平均生长速率[62.30 mg/(L·d)]分别提高24.32%和31.66%;脂类产量(194.91 mg/L)和平均合成速率[13.21 mg/(L·d)]分别提高45.59%和45.64%,脂肪酸组成中C16:0和C18:1分别上升20.56%和19.27%;最高叶绿素a和虾青素浓度达到3.89 mg/L和4.37 mg/L,分别提高20.43%和35.71%。表明TD16与雨生红球藻混合培养能够促进雨生红球藻的生长、提高脂类产量。更多还原

【Abstract】 Haematococcus pluvialis was seen as an ideal raw material for biodiesel for the main FAME compositions are palmitic acid,oleic acid linolic acid,and less unsaturated double bonds,etc.However the relatively high cost restricted the large scale industrialized production of H.pluvialis.This investigation focused on increasing the growth rate and the lipid content of H.pluvialis respectively enhance the yield per unit and lower the processing cost for the biomass per unit,accordingly increased the economical efficiency and the sustainability of the process.Therefore we mixed cultivated H.pluvialis and Paecilomyces lilacinus TD16(hereinafter referred to as TD16)in this investigation to study the influence of fungus to the growth of microalgae and the synthesis of lipid,the main results are show as follows.TD16 extracellular proteins and polysaccharide can both promote the growth of the biomass and the content of lipid of H.pluvialis,further the degree of the promotion of TD16extracellular proteins to frustule was larger than polysaccharide,meanwhile we found that TD16 extracellular proteins pull a bacteriostasis effect to Staphylococcus aureus,Bacillus subtilis and Escherichia coli.The TD16 suspension was inoculated to BBM substrate containing H.pluvialis extracellular organic carbon,the results showed that the TD16biomass increasing constantly,simultaneously the extracellular organic carbon in the substrate was decreasing constantly,these indicated that the extracellular organic secreted by the H.pluvialis can be utilized.Through single factor experiment we acquired the optimal cultivation conditions:nutritional modes,inoculation ratio of H.pluvialis and TD16(initial concentrations were1×10~7cells/m L and 1.36×10~7cells/m L),concentration of sodium nitrate,medium p H,light intensity and culture temperature were photosynthetic autotrophy,5:1,0.35 g/L,7,70mol/(m~2.s)and 25℃,respectively.The effects and mechanisms of TD16 on the growth and lipid synthesis of H.pluvialis were systematically studied.Compared with the single culture of H.pluvialis,the cell dry weight(0.99 g/L)and the average growth rate[62.30 mg/(L.d)]of the mixed culture of H.pluvialis-TD16 increased by 24.32%and 31.66%,respectively.The lipid yield(194.91mg/L)and the average synthesis rate[13.21 mg/(L.d)]were increased by 45.59%and45.64%,respectively,and the fatty acid composition of C16:0 and C18:1 increased by20.56%and 19.27%,respectively.The highest concentrations of chlorophyll a and astaxanthin were 3.89 mg/L and 4.37 mg/L,which increased by 20.43%and 35.71%respectively.The results showed that the mixed culture of TD16 and H.pluvialis can promote the growth of H.pluvialis and increase the lipid yield.更多还原