高产油脂微藻的筛选及其对生活污水脱氮除磷能力的研究

发布时间：2018-03-23 02:16

本文选题：能源微藻　切入点：氮磷去除　出处：《山东大学》2015年硕士论文　论文类型：学位论文

【摘要】：随着能源的消耗与需求越来越大,环境问题越来越引起人们的关注与担忧,此外逐渐涨高的油价也引起人类的恐慌,因此找到一种可替代的新能源迫在眉睫。作为第三代生物能源的原材料—微藻,由于其具有生长速率快,油脂含量高,不占农耕面积等竞争性优势一直以来是研究的热点。并且,微藻制备的生物柴油是一种可生物降解,可再生,无毒,碳中和的环境友好型能源。因此,利用生物柴油取代化石燃料是大势所趋。然而微藻产油要实现大规模工业化生产,必须要解决产率低,成本高这两大瓶颈问题,即要筛选高产油脂微藻,降低生产成本。针对成本高这一问题,研究者提出利用污水作为微藻培养的替代培养基,微藻生长的同时吸收其中的氮磷等营养物质,不仅起到污水处理的作用,同时积累生物质。这一概念的提出实现了污水处理系统向生产系统的转化,兼具环境效益与经济效益。本文从济南市当地水体中分离纯化微藻,从中初步筛选出环境适应性强及高产油脂微藻四株。利用人工配制的生活污水,考察四株微藻在人工配水中的生长状况,氮磷去除能力及油脂积累特性,最终从中选出性能最佳的一株栅藻。循环利用校园污水作为Scenedesmus SDEC-13的替代培养基,考察其对于校园污水的利用及处理情况,并研究了该栅藻的油脂及蛋白积累特性。本文得到的主要研究结论如下：(1)通过平板分离法及96孔板法筛选获得纯藻株4株。透射电镜分析显示四株藻藻体内含有大量油脂滴,因此其可作为富油微藻来研究。对其进行形态学及分子生物学学上的分析鉴定,最终给四株藻分别命名为Chlorella sp. SDEC-10, Chlorella sp. SDEC-11, Scenedesmus sp. SDEC-12, Scenedesmus sp. SDEC-13。此外,4株藻已被保存至中国科学院淡水藻种库,编号分别为：]FACHB-1765, FACHB-1767, FACHB-1766, FACHB-1768。其中Scenedesmus sp. SDEC-12， Scenedesmus sp. SDEC-13也于中国典型培养物保藏中心(武汉)予以保藏,保存编号分别为：CCTCC NO: M 2014497和CCTCC NO: M 2014498。(2)四株微藻的环境适应性强,能够在人工配制的生活污水中迅速生长,并且获得比BG-11培养基中更高的生物量。四株微藻在人工配水中获得了高的油脂产量,均高于25%。四株藻在人工配水中生长的同时,吸收其中的氮磷营养物质,对于污水中的氮磷有明显的去除效果,其中磷的去除率几乎为100%,氮的去除率相对低。(3)综合比较四株藻在人工配制的生活污水中的生长状况,油脂积累特性以及氮磷去除能力, Scenedesmus SDEC-13获得最大的生物量产率(68.92 mg/L·d),其对于氨氮的去除效率最高达65%,磷去除率达100%,油脂产率高,并且其脂肪酸甲酯组成分析表明更适合作为生物柴油的原料,因此,四株微藻中SDEC-13是优势藻种。(4)利用未经灭菌、稀释等前处理而只经六层纱布过滤的校园污水作为替代培养基,培养Scenedesmus SDEC-13。校园污水可以被循环使用两次来满足SDEC-13的生长需求,同时积累油脂、合成蛋白。两次循环利用校园污水培养SDEC-13获得的油脂产量较高,均在25%以上；脂肪酸组成分析表明,校园污水中培养的SDEC-13中脂肪酸组分更适合生产高品质的生物柴油。蛋白含量均在30%以上,氨基酸组成分析显示获得的蛋白品质较高。校园污水经过两次利用后其中的氮磷得到有效去除,最终出水可以满足市政污水处理厂污染物排放标准的一级A标准而达标排放。
[Abstract]:With the energy consumption and demand more and more, the environmental problems caused by more and more people's attention and concern, also gradually rose high oil prices have caused human panic, so to find a new alternative energy sources is imminent. As a raw material - microalgae third generation biofuels, due to its fast growth rate, oil content high, does not account for the farming area of competitive advantage has been the hotspot of research. Also, microalgae for biodiesel preparation is a biodegradable, renewable, non-toxic, environmentally friendly energy carbon neutral. Therefore, the use of bio diesel to replace fossil fuels. However, microalgae oil is represent the general trend to achieve large-scale industrial production. We must solve the low yield, high cost of these two major bottlenecks, namely to screening of high lipid microalgae, reduce the production cost. In order to solve the problem of high cost, the researchers proposed the use of sewage for Alternative medium for culturing microalgae and microalgae growth and absorption of nitrogen and phosphorus and other nutrients, not only play the role of sewage treatment, while the accumulation of biomass. The concept of the sewage treatment system to achieve the transformation of the production system, both environmental benefits and economic benefits. In this paper, the separation and purification of microalgae from the local water Ji'nan City, from the initial screening of environmental adaptability and high yield. Four strains of oleaginous microalgae with synthetic sewage of four microalgae growth conditions on the artificial water, nitrogen and phosphorus removal capacity and oil accumulation characteristics, finally choose the optimal performance of a strain of Scenedesmus cycle as a replacement for the Scenedesmus SDEC-13. The medium used for the use of the campus sewage, and treatment of campus sewage, and studied the oil and protein accumulation characteristics of Scenedesmus. Mainly studies the The method is as follows: (1) and 96 well microtiter plate screening to obtain pure strains 4 strains by plate separation. TEM analysis showed that four strains of algae body contains a lot of fat drops, so it can be used as oil rich research. Study on the identification and analysis of morphology and molecular biology of the final to four strains of algae named Chlorella sp. SDEC-10, Chlorella sp. SDEC-11, Scenedesmus sp. SDEC-12, Scenedesmus sp. SDEC-13., in addition, 4 strains of algae have been preserved to Chinese Academy of freshwater algae species pool, numbered:]FACHB-1765, FACHB-1767, FACHB-1766, FACHB-1768. and Scenedesmus sp. SDEC-12, Scenedesmus sp. SDEC-13 in Chinese (Wuhan type culture collection center to be preserved, preserved) numbered: CCTCC NO: M 2014497 and CCTCC NO: M 2014498. (2) of four strains of microalgae and environmental adaptability, can be formulated in artificial sewage In the rapid growth, and cultivation of higher biomass medium than BG-11. Four strains of microalgae oil with high yield in synthetic water were higher than those of 25%., four strains of algae in the water and artificial growth, absorption of nitrogen and phosphorus nutrients, for removal of nitrogen and phosphorus in the sewage has obvious among them, the phosphorus removal rate is almost 100%, the nitrogen removal rate is relatively low. (3) the growth status of comprehensive comparison of four strains of algae in artificial sewage, oil accumulation and characteristics of nitrogen and phosphorus removal ability of Scenedesmus SDEC-13 to obtain the maximum biomass yield (68.92 mg/L d), the ammonia nitrogen the removal efficiency of up to 65%, the phosphorus removal rate was 100%, the oil yield is high, and its methyl fatty acid composition analysis showed that more suitable as raw material for biodiesel, therefore, four strains of microalgae in SDEC-13 is the dominant species. (4) use without sterilization, dilution and pretreatment Only through six layers of gauze filter campus sewage as substitute of medium, culture Scenedesmus SDEC-13. campus sewage can be recycled two times to meet the growing demand of SDEC-13, while the accumulation of fat, protein synthesis. The two recycling of campus sewage grease culture high yield SDEC-13, were more than 25%; analysis shows that fat acid, fatty acid composition of biodiesel is more suitable for the production of high quality cultivation of campus sewage in SDEC-13. The protein content was above 30%, the amino acid composition analysis showed that the protein quality obtained higher. Campus sewage after two times after the use of nitrogen and phosphorus which can be efficiently removed, the final effluent can meet the discharge standard of municipal sewage treatment plant the level of A standard and pollutant discharge standards.

【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：X799.3;X173

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