盐藻和小球藻富集铬（Ⅲ）的优化培养条件研究
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摘要
研究以蛋白核小球藻和杜氏盐藻为材料，盐藻设定7个添加Cr~(3+)浓度处理，小球藻设定8个添加Cr~(3+)浓度处理，测定了不同Cr~(3+)浓度下微藻的生物量(细胞密度)、微藻对Cr~(3+)的富集量、叶绿素a、β-胡萝卜素、蛋白质及可溶性糖含量；选择其中对微藻生长和富集较优的添加Cr~(3+)浓度，比较了Cr~(3+)的不同添加方式对微藻生物量及富集量的影响；进行了两种微藻的三因素三水平正交试验，测定了不同组合的生物量和富集量；并探讨了最佳组合条件下的Cr~(3+)富集动态过程；分析了生物富集对微藻细胞分子官能团结构的影响。结果表明，培养液的Cr~(3+)浓度为1.0mg·L~(-1)时，可以提高小球藻的生物量、叶绿素a、蛋白质和可溶性糖的含量。小球藻富集Cr~(3+)的适宜添加条件为：在接种第1天至第5天期间分批次添加Cr~(3+)，其最终加入是1.0mg·L~(-1)，收获时测小球藻的生物量及富集量分别是2.25mg·mL~(-1)和82.89μg·g~(-1)，小球藻对Cr~(3+)的富集效率为88.7％，与一次性添加相比较，小球藻的富集量提高了28.2％。小球藻获得最大生物量和富集量的优化组合为：温度30℃，pH7，N3.0g·L~(-1)，P0.08g·L~(-1)，此时的生物量为2.38mg·mL~(-1)，富集量为85.35μg·g~(-1)。Cr~(3+)浓度为0.2mg·L~(-1)时，可以提高盐藻的生物量、β-胡萝卜素、蛋白质、可溶性糖含量。盐藻富集Cr~(3+)的适宜添加条件为：在接种第1天至第8天期间分批次添加Cr~(3+)，其最终加入是0.2mg·L~(-1)，收获时测盐藻的生物量及富集量分别是0.93mg·mL~(-1)和40.23μg·g~(-1)，盐藻对Cr~(3+)的富集效率为90.6％，与一次性添加相比较，盐藻的富集量提高了32.6％。盐藻获得最大生物量和富集量的优化组合为：温度30℃，pH7，盐度70‰，此时的生物量为0.98mg·mL~(-1)，富集量为42.77μg·g~(-1)。
In this paper, we used Chlorella pyrenoides and Dunaliella salina as experimental material to assess the optimal cultivation conditions of Cr enrichment by the two alga species through cultivation experiments by Cr3+ addition in the medium. Chlorella pyrenoides was cultivated under the 7 different concentrations of Cr3+ and Dunaliella salina was cultivated under the 8 different concentrations of Cr3+. We measured biomass (cell density) of the two algae, the amount of Cr3+ enrichment, contents of chlorophyll-a, β -carotene, intracellular protein and carbohydrate in the different Cr3+ addition treatments. Based on these experiments, we used orthogonal experiment with three factors and three levels, analyzed the effects of different methods of Cr3+ addition on algal biomass and Cr enrichment and discussed the dynamic processes of Cr3+ enrichment under the optimal combination conditions. In addition, the effects of Cr3+ enrichment on the algal cell molecular functional group structure were also investigated. The
result of this study indicated that Cr3+ significantly increased the biomass of Chlorella pyrenoides and its contents of chlorophyll-a, β -carotene, protein and carbohydrate at the Cr3+ concentration of Img ?L"-1. The optimal concentration of Cr3+ was Img ?L-1 that should be added day by day to the medium during the initial 5d of culture. At this condition, harvested biomass and Cr enrichment quantity of Chlorella pyrenoides were 2.25mg ?mL-1 and 82.89 *g ?g-1, respectively, with an enrichment efficiency of 88.7%. The optimal combination of temperature, pH and N, P concentration depended on the experimental intention. To improve the growth and Cr3+ enrichment of Chlorella pyrenoides, the optimal combination of temperature, pH and N, P concentration is 30 ℃ in temperature, 7 in pH, 3g ?L-1 in N concentration and .08g 'L-1 in P concentration, with the highest biomass of 2.38mg 'mL-1 and the highest enrichment quantity of 85.35 * g ?g-1. The concentration of 0.2mg ?L-1 in Cr3+ can significantly increas
ed the biomass of Dunaliella salina and its contents of chlorophyll-a, 3 -carotene, protein and carbohydrate. The optimal concentration of Cr3+ was 0.2mg ?L-1 that should be added day by day to the medium during the initial 8d of culture. At this condition, harvested biomass and Cr enrichment quantity of Dunaliella salina were 0.93mg ?mL-1 and 40.23 * g ?g-1, respectively, with an enrichment efficiency of 90.6%. The optimal combination is 30℃ in temperature, 7 in pH and 70 %o in salinity because the highest biomass of 0.98mg ?mL-1 and the highest enrichment quantity of 42.77 μ g ?g-1.
In this paper, we used Chlorella pyrenoides and Dunaliella salina as experimental material to assess the optimal cultivation conditions of Cr enrichment by the two alga species through cultivation experiments by Cr3+ addition in the medium. Chlorella pyrenoides was cultivated under the 7 different concentrations of Cr3+ and Dunaliella salina was cultivated under the 8 different concentrations of Cr3+. We measured biomass (cell density) of the two algae, the amount of Cr3+ enrichment, contents of chlorophyll-a, β -carotene, intracellular protein and carbohydrate in the different Cr3+ addition treatments. Based on these experiments, we used orthogonal experiment with three factors and three levels, analyzed the effects of different methods of Cr3+ addition on algal biomass and Cr enrichment and discussed the dynamic processes of Cr3+ enrichment under the optimal combination conditions. In addition, the effects of Cr3+ enrichment on the algal cell molecular functional group structure were also investigated. The
result of this study indicated that Cr3+ significantly increased the biomass of Chlorella pyrenoides and its contents of chlorophyll-a, β -carotene, protein and carbohydrate at the Cr3+ concentration of Img ?L"-1. The optimal concentration of Cr3+ was Img ?L-1 that should be added day by day to the medium during the initial 5d of culture. At this condition, harvested biomass and Cr enrichment quantity of Chlorella pyrenoides were 2.25mg ?mL-1 and 82.89 *g ?g-1, respectively, with an enrichment efficiency of 88.7%. The optimal combination of temperature, pH and N, P concentration depended on the experimental intention. To improve the growth and Cr3+ enrichment of Chlorella pyrenoides, the optimal combination of temperature, pH and N, P concentration is 30 ℃ in temperature, 7 in pH, 3g ?L-1 in N concentration and .08g 'L-1 in P concentration, with the highest biomass of 2.38mg 'mL-1 and the highest enrichment quantity of 85.35 * g ?g-1. The concentration of 0.2mg ?L-1 in Cr3+ can significantly increas
ed the biomass of Dunaliella salina and its contents of chlorophyll-a, 3 -carotene, protein and carbohydrate. The optimal concentration of Cr3+ was 0.2mg ?L-1 that should be added day by day to the medium during the initial 8d of culture. At this condition, harvested biomass and Cr enrichment quantity of Dunaliella salina were 0.93mg ?mL-1 and 40.23 * g ?g-1, respectively, with an enrichment efficiency of 90.6%. The optimal combination is 30℃ in temperature, 7 in pH and 70 %o in salinity because the highest biomass of 0.98mg ?mL-1 and the highest enrichment quantity of 42.77 μ g ?g-1.
引文
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[2] 陈小霞，梁世中，吴振强，岳振峰．异养小球藻生物富集Cr~(3+)的研究．食品与发酵工业，2001，(27)：33-36．
[3] 陈小霞，吴振强，梁世中．藻类对微量元素的生物富集及其机理探讨．食品与发酵工业，1997，25(4)：56-60．
[4] 陈永秉．数理统计浅说．北京：农业出版社，1993：223-230．
[5] 范晓，王孝举．海藻中的碘．海洋科学，1994(4)：16-20．
[6] 古玉环，陈军．盐生杜氏藻(Dunaliella salina)的生物学特性与培养研究．西北师范大学学报(自然科学版)，1995，31(4)：52-55．
[7] 郝涤非．螺旋藻的特性利用与培养．生物学教学，1999，24(3)：39—44．
[8] 浩云涛，李建宏，潘欣．椭圆小球藻(CHorelln ellipsoidea)对4种重金属的耐受性及富集．湖泊科学，2001．13(2)：158-162．
[9] 贺超兴．盐藻的生物学特性与开发利用．生物学通报，1998，33(2)：12-16．
[10] 姜健国，周世水，姚汝华．盐藻的诱变育种及光合反应器培养的初步研究．海洋科学，2001，25(1)：44-46．
[11] 李乐农，郭宝江．螺旋藻的培养及其分子生物学研究概况．植物学通报，1998，15(增刊)：72-76．
[12] 李平，梁世中．微量元素铬作为功能食品因子的研究进展．食品与发酵工业，2000，27(1)：74-77．
[13] 李师翁，李虎乾，张建军．小球藻大规模培养研究的进展．植物学通报，1998，15(4)：45-50．
[14] 李师翁，李虎乾．植物单细胞蛋白质源-小球藻开发利用研究的现状．生物技术，1997，7(3)：45-48．
[15] 李志勇，郭祀远，李琳，蔡妙颜．藻类对微量元素的生物富集及其应用．微生物学通报，1997，24(6)：368-369．
[16] 李志勇，郭祀远，李琳，蔡妙颜．藻类富集微量元素的机理研究．华南理工大学学报(自然科学版)，1998，26(2)：33—37．
[17] 李志勇，郭祀远，李琳．功能性高铬(Ⅲ)螺旋藻的研制．中国海洋药物，1999，(4)：14-18．
[18] 李志勇，李元广，郭祀远，李琳．钝顶螺旋藻生物富集Cr~(3+)影响因素的研究．生物工程学报，2000，16(1)：108-112．
[19] 李玉山，崔征，殷军．七种马尾藻属海藻碘及微量元素的含量测定．中国海洋药物，1996，(4)：35-37．
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