【作者】 王志忠；

【导师】 刘果厚；

【作者基本信息】 内蒙古农业大学 ， 野生动植物保护与利用， 2015， 博士

【摘要】 鄂尔多斯鄂托克地区是国内钝顶螺旋藻的优势产区,然而该地区近二十来钝顶螺旋藻养殖的培养基配方陈旧,资源浪费严重；采收干燥及藻蓝蛋白提取工艺粗略；藻粉灰分、砷含量偏高；藻种保存技术落后等问题突出,严重制约该地区螺旋藻产业的长足发展。在这样的背景下,本文以该地区钝顶螺旋藻为主要研究对象,系统开展了养殖原料利用规律、砷富集规律、清洗控水干燥工艺、藻蓝蛋白提取工艺、藻种越冬形态及保存方法几个方面的研究,建立适合该地区钝顶螺旋藻的生产、加工以及保存的关键技术体系,旨在为钝顶螺旋藻的高效、优质生产提供技术参考和理论依据。主要研究结论如下：1.通过单因素及正交试验,在鄂托克旗气候环境下,该地区钝顶螺旋藻基础培养液中NaHCO3、NaNO3、H3PO4、KCl、MgSO4·7H2O、FeSO4·7H2O的最佳添加浓度分别为63 g/L、1.0g/L、0.06 g/L0.5 g/L、0.1g/L、0.007 g/L,引进钝顶螺旋藻分别为21 g/L、1.5 g/L、0.12 g/L、0.5 g/L、0.1 g/L、0.007 g/L;单位收藻周期后2藻种需要补加6种原料的浓度分别为06 g/L、0.06 g/L、0.01 g/L、0.04 g/L、0.015 g/L、0.002 g/L和0.4 g/L、0.04 g/L、0.007 g/L、0.03 g/L、0.01g/L、0.0015 g/L。总体上,鄂尔多斯高原碱湖钝顶螺旋藻的生长和产量都优于引进钝顶螺旋藻。每天添加0.05 g/LL的NaNO3以及0.02 g/L的CO(NH2)2或008g/L的NH4HCO3可显著促进螺旋藻生长；在封闭养殖环境中,循环使用当地深层地下水可补充螺旋藻生长所需的微量营养起到增加产量且降低成本的作用；添加Ca2+、A5液和B6液对螺旋藻生长的影响不显著；当藻液中NaCl、Na2CO3的浓度分别超过10g/L、20 g/L时,螺旋藻生长不良,需要更换部分或全部培养液。2.藻液的砷来源于地下水和NaHCO3,比例分别占72%和26%,螺旋藻养殖各个时期的藻液砷浓度基本保持稳定；蒸馏水辅助清洗藻泥可显著降低藻粉中的砷含量,培养液砷含量小于0.32 mg/L时对螺旋藻的生长和产量的影响不显著；在不同砷含量的培养液中,藻液中的砷含量与藻粉砷含量呈显著正相关,总体上,鄂尔多斯高原碱湖钝顶螺旋藻的生长和产量优于引进藻种,且砷含量显著低于引进藻种；通过单细胞紫外诱导,培养获得单藻丝,然后在不同砷浓度的营养液中进行抗砷培养,选定了2个生长速率较快且砷含量富集较低的鄂尔多斯高原钝顶螺旋藻品系。3.螺旋藻采收过程中,蒸馏水清洗或0.00001mol/L勺HC或0.0001%的H3PO4浸泡再辅以地下水清洗,藻粉灰分可降到7%以下；常温下,随着藻泥控水时间的延长藻体类胡萝卜素含量、叶绿素a含量和SOD活性降低；清洗干净的藻泥尽快干燥或及时冷藏保鲜贮存均可显著降低螺旋藻体上述物质的损耗；高温喷雾干燥较晒干、烘干可保持螺旋藻较高的类胡萝卜素、叶绿素α、SOD及蛋白质含量。4.在藻蓝蛋白提取过程中,保持较高的藻蓝蛋白纯度且降低提取成本的盐析硫酸铵饱和度为40-50%,鄂尔多斯高原碱湖钝顶螺旋藻藻蓝蛋白的纯度最大值为引进藻种的1.67倍,但两藻种的提取率没有差异；鄂尔多斯高原碱湖钝顶螺旋藻藻蓝蛋白对温度敏感性显著高于引进藻种,55℃是保持较高藻蓝蛋白活性的临界温度；不同pH值的提取液对藻蓝蛋白的纯度影响显著,pH值5—6时,两藻种藻蓝蛋白纯度均最大。5.鄂尔多斯高原碱湖钝顶螺旋藻是以营养体(即完整或断裂为短段的藻体)在天然碱湖中越冬；且5℃弱光保存鄂尔多斯高原碱湖钝顶螺旋藻藻种的效果最佳。更多还原

【Abstract】 There are many serious problems in many respects such as antiquated medium formulation, serious waste of resources, the rough process of harvest and Phycocyanin extraction, the more content of ash and As and uncultured methods saving Spirulina species, which severely restrict the development of Spirulina industry in Etuoke Qi region though it is the main Spirulina region owning advantage in China. The technical parameters of the nutrient utilization, arsenic enrichment, cleaning, removing water and drying in the harvesting process, extracting Phycocyanin, the Spirulina platensis morphology at low temperatures and preservation methods were studied, in order to establish the key technology system suitable for this area for the production, processing and preservation of Spirulina platensis to provide technical reference and theoretical basis for high-yield, efficient, high-quality production of Spirulina platensis. The main conclusions are as follows:1. Under the same culturing conditions in Etuoke Qi, the results of single factor experiment and orthogonal experiment showed the optimum culturing consentration of NaHCO3,NaNO3, H3PO4,KCl, MgSO4·7H2O、FeSO4·7H2O to Spirulina platensis from Ordos were 6.3 g/L,1.0 g/L,0.06 g/L,0.5 g/L,0.1 g/L、0.007 g/L respectively, while 2.1 g/L,1.5 g/L,0.12 g/L,0.5 g/L,0.1 g/L、0.007 g/L for the introduced species. The average supplemented consentration of NaHCO3、NaNO3、 H3PO4、KCl、MgSO4·7H2O、 FeSO4·7H2O were 0.6 g/L,0.06 g/L,0.01 g/L,0.04 g/L,0.015 g/L、0.002 g/L to culture Spirulina platensis from Ordos after harvested for each time, while 0.4 g/L,0.04 g/L, 0.007 g/L,0.03 g/L,0.01 g/L、0.0015 g/L for the introduced species. The growth and yield of Spirulina platensis from Ordos were better than the introduced species. The growth of Spirulina platensis from Ordos was promoted obviously by fertilizing with 0.05 g/L NaNO3 and 0.02 g/L CO(NH2)2 or 0.08 g/L of NH4HCO3 per day. Yield of Spirulina platensis increased, the production cost reduced and trace elements and materials were saved by circularly using groundwater in closed culturing facilities. The growth of Spirulina platensis was not affected when without Ca2+, A5 and B6 liquid. The concentration of NaCl and Na2CO3 were higher than 10 g/L and 20 g/L could inhibit the growth of Spirulina platensis, so the culture medium should be replaced partially or completely.2. Arsenic content in the culture medium was came from the groundwater, NaHCO3 and NaNO3, and the percentage rate was 72% and 26% respectively. The arsenic content could be correspondingly kept in a stable state in each period of the cultivation. The arsenic content was reduced by cleaning with distilled water. The growth and production of two species were not inhibited when the arsenic in the culture medium was lower than 0.32 mg/L; the production of Spirulina platensis from Ordos was higher than that of introduced species, arsenic content of Spirulina powder of Ordos’s was obviously lower than that of introduced species under different arsenic concentratio.The arsenic content of the culture medium was positive correlation with the arsenic content of the Spirulina powder. Single-celled Spirulina platensis was induced mutation by radiation, then cultured in the nutrient solution of different arsenic concentrations, two strains of Spirulina platensis were selected which not only grew fast, but also remain the lowest content.3. Ash content of Spirulina powder was no more than 7% by cleaning with distilled water or acid soaked (0.00001 mol/L HCl or 0.0001% H3PO4) then washed with groundwater. The content of Carotenoids and Chlorophyll a and SOD activity decreased with the extension of exposure time at room temperature. Spirulina mud should be dried as soon as possible or placed in cold storage equipment promptly to make the loss of the content of Carotenoids and Chlorophyll a and SOD activity of Spirulina platensis slow down. The more effective way to dry Spirulina making the content of Carotenoids, Chlorophyll a and protein and SOD activity, the larger was spray drying rather than drying under sunlight or oven drying.4. The optimum concentration of (NH4)2SO4 was 40%-50% for protein extraction of Phycocyanin and decrease the extraction cost. The Phycocyanin purity of Spirulina platensis from Ordos was 1.67 times of the imported species, extraction rate was close for two species.The purity of Phycocyanin for Spirulina platensis from Ordos was significantly more sensitive to temperature than the introduced species; the working temperature controlled less than 55 ℃, the higher Phycocyanin activity could be maintained; pH value had significant influence to the Phycocyanin purity, the value of Phycocyanin purity was highest when pH value in 5-6.5. Spirulina platensis from the natural alkali lake in the Ordos plateau can withstand the cold winter with the vegetative form or its short broken segment under the low temperature. Spirulina platensis from Ordos can well survived under 5℃ and weak light conditio.更多还原