饵料型微藻新品系培育及工程化高效生产技术

李凌

摘 要：建立以主要饵料微藻的可持续育种技术体系和育种群体，培育高产、抗逆新品系，突破微藻培养广鉴技术，构建成熟的微藻饵料工程化生产技术与工艺为目标。通过对18S rDNA、rbcL基因和ITS区序列的测定及系统进化分析，建立了微藻种间及株系间分子鉴别技术；对多株饵料微藻的生长及适应性进行了比较研究，筛选出了适应不同光强的雨生红球藻藻株，抗逆性强的小球藻、三角褐指藻，耐高温的纤细角毛藻藻株及盐藻、扁藻等多株优良饵料微藻，为规模化培养提供了良好材料。研究了微藻高密度培养中的生长指标和适应机制，并在此基础上针对雨生红球藻、微拟球藻、三角褐指藻、小球藻等多种饵料微藻研究了不同培养条件及添加物对其生长及营养物质积累的影响，建立了多种饵料微藻的高密度培养技术，其中微拟球藻在气升式螺旋管道（光径1.2 cm）光生物反应器中培养时，最高密度可达15 g/m2，比目前常规技术提高了3～4倍。针对饵料微藻的特点，设计了规模大小递增的光生物反应器，建立了敌害生物综合防御的方法和技术体系，有效降低污染几率；克服了传统封闭培养中氧气解析、二氧化碳补偿、光线分布、物料均质、细胞贴壁等难题，实现了主要参数温度、pH值和溶解氧等

数据的在线检测和自动控制。通过实验室100mL-500mL-，户外200L-多组并联柱状光生物反应器逐级扩大培养，建立了多种饵料微藻工程化生产工艺，建设了2个产业化基地，实现300 t体积的规模培养。以三角褐指藻、小球藻和牟氏角毛藻3种饵料型微藻为对象，通过在现有液固分离的小型设备上进行浓缩效果实验，确定了生产型的浓缩设备并针对以上3种不同微藻的特性，进行了微藻浓缩工艺的研究，建立了不同微藻的浓缩工艺。优化了微藻产品（藻膏）低温保存技术，形成微藻产品浓缩与保存技术规模和规范。

关键词：饵料型微藻 筛选 高效培养 光生物反应器

Abstract：The aim of the program is to establish sustainable breeding technological system and group of the main bait microalgae， cultivate high-yielding， resistant strains， break through the identification technology， and build large-scale production technology of bait microalgae. The molecular identification technology of microalgae was established by phylogenetic analysis of 18S rDNA， rbcL gene and ITS sequences. Based on the comparison of growth and adaptability of multiple microalgae strains， Haematococcus pluvialis adapted to different light intensities， Chlorella sp. and Phaeodactylum tricornutum with strong stress resistance，Chaetoceros gracilis resistant to high temperature， and other good strains （e.g. Dunaliella salina and Tetraselmis sp.） were selected to provide materials for large-scale culture. The growth index and adaptation mechanism of microalgae in high density cultivation were studied， and through researching the effect of culture conditions and additives on the growth and nutrient accumulation of several microalgae （e.g. H. pluvialis， Nannochloropsis sp.，P.tricornutum，Chlorella sp.），a high-density cultivation technology were established， by which the biomass of Nannochloropsis sp.increased 3～4 times，and reached up to 15 g/m2 in airlift helical pipes photobioreactors （diameter 1.2 cm）. Based on the characteristics of bait microalgae，photobioreactors with different sizes were designed， pollution risk was efficiently reduced by establishing technology for predators extermination， the traditional problem in closed culture， such as oxygen release，carbon dioxide compensation， light distribution， material homogenization， and cells attachment were overcome， and the main parameters，such as temperature， pH and dissolved oxygen data were detected online and controlled automatically.A large-scale production technology of microalgae was established through enlargement culture step by step using 100 mL to 500 mL and 200 L column photobioreactor，and two industrialization bases were built to meet the cultivation scale of 300 tons. Based on the characteristics of P.tricornutum，Chlorella sp. and Chaetoceros muelleri， concentration technology for different microalgae was established through experiment on small liquid-solid separation equipment. Cryopreservation technology of microalgae products （algae paste） was optimized， and the technical specification for concentration and preservation of microalgae products was formed.

Key Words：Bait microalgae；Screen；Efficient culture；Photobioreactor

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