海洋微藻在利用碳酸盐离子溶液处理烟道气CO2中的耐受机理研究

The core regulatory mechanism of algae absorbing carbon dioxide was CO2 concentrating mechanism ( CCM ), most studies were focusing on single component function of CCM during algae absorbing inorganic carbon. There were no reports on mechanism of algae tolerating high NaHCO3 which was converted from Na2CO3 using CO2 of flue gas. We acquired high-tolerant bicarbonate marine algae through screening seawater samples, then investigated the response changes of the key component of inorganic carbon transport pathways in CCM during tolerating high bicarbonate from the transporter protein expression, carbonic anhydrase expression and enzyme activity levels, and enzyme gene differences, to understand in-depth the marine inorganic carbon transport mechanism. Afer that the inorganic carbon utilization efficiency were evaluated using 14C isotope tracer techniques, algae metabolite were extracted using Sequential Subcritical Hydrothermal Extraction (SSHTE) Technology and identified the high-value products combined with gas chromatography - mass spectrometry (GC/MS), the optimal conditions coupling the growth rate, the carbon use efficiency and high additional value product contents were explored, finally it provided theoretical principle for existing algae genetic engineering and more economical effective ways of capturing and using of the greenhouse gas to realize the harmonious of human, environment, resource.

微藻吸收二氧化碳的核心调控机制是碳浓缩机制(CCM)，在微藻吸收无机碳过程中大部分研究集中在CCM 机制单组分的功能和机理方面，目前没有关于海水藻通过Na2CO3 吸收利用烟道气中CO2 转化为高浓度NaHCO3 离子溶液耐受机理的报道，我们通过筛选获得耐受高浓度碳酸氢钠离子溶液的海水藻，对其无机碳转运途径中的关键组分分别从转运蛋白表达量、碳酸酐酶表达量和酶活水平、及酶的基因差异上揭示海藻耐受高浓度碳酸氢钠过程中CCM 组分响应变化规律，深入理解海水藻无机碳转运的调控机制；采用14C 同位素示踪技术对其碳的利用效率进行评价，利用次临界热解萃取（SSHT）海藻合成物，结合气相色谱- 质谱鉴定高附加值产品，探索生长速率、碳利用效率和高附加值产品含量三者耦合的最优条件；为基因工程改造现有藻种提供理论基础，为捕获和利用温室效应气体提供更加经济有效的途径，实现人类、环境、资源的和谐统一。

微藻吸收二氧化碳的核心调控机制是碳浓缩机制(CCM)，目前没有关于海水藻通过Na2CO3吸收利用烟道气中CO2转化为高浓度NaHCO3离子溶液耐受机理的报道，我们通过筛选获得一株耐受高浓度碳酸氢钠离子溶液的海水藻HTBS，经鉴定与杜氏盐藻较为相近。HTBS具有较强的温度适应性(7 – 30 °C)，能够在含有30% CO2或者70 g L−1 NaHCO3的培养基中生长，在添加25g·L−1NaHCO3的培养基中快速生长。对其无机碳转运途径中的关键组分分别从转运蛋白表达量、碳酸酐酶表达量和酶活水平、及酶的基因差异上揭示海藻耐受高浓度碳酸氢钠过程中CCM组分响应变化规律，经过免疫印迹研究碳酸氢钠处理条件下HTBS藻株和非耐受碳酸氢钠藻株微拟球藻α-CAs表达，结合碳酸酐酶专一抑制剂AZ和EZ确认α-CAs定位，表明CAH1和CAH2是HTBS耐受高HCO3-的必要酶蛋白。利用iTRAQ技术对NaHCO3胁迫和正常生长细胞的差异表达蛋白检测，经蛋白组学分析共得到196个差异表达蛋白，其中上调84个，下调112个，经过Gene Ontology和KEGG pathway分析，差异蛋白主要富集在能量代谢与离子运输过程。通过以上分析深入理解了海水藻无机碳转运的调控机制，为基因工程改造现有藻种提供理论基础，为捕获和利用温室效应气体提供更加经济有效的途径。

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