Bernal Oscar I, Mooney Charles B, Flickinger Michael C
Department of Chemical and Biomolecular Engineering, North Carolina State University, 911 Partners Way, Raleigh, North Carolina, 27695.
Biotechnol Bioeng. 2014 Oct;111(10):1993-2008. doi: 10.1002/bit.25280. Epub 2014 Jul 18.
We describe a latex wet coalescence extrusive coating method that produces up to 10-fold specific photosynthetic rate enhancements by nitrate-limited non-growing cyanobacteria deposited onto paper, hydrated and placed in the gas-phase of small tube photobioreactors. These plant leaf-like biocomposites were used to study the tolerance of cyanobacteria strains to illumination and temperature using a solar simulator. We report sustained CO2 absorption and O2 production for 500 h by hydrated gas-phase paper coatings of non-growing Synechococcus PCC7002, Synechocystis PCC6803, Synechocystis PCC6308, and Anabaena PCC7120. Nitrate-starved cyanobacteria immobilized on the paper surface by the latex binder did not grow out of the coatings into the bulk liquid. The average CO2 consumption rate in Synechococcus coatings is 5.67 mmol m(-2) h(-1) which is remarkably close to the rate reported in the literature for Arabidopsis thaliana leaves under similar experimental conditions (18 mmol m(-2) h(-1) ). We observed average ratios of oxygen production to carbon dioxide consumption (photosynthetic quotient, PQ) between 1.3 and 1.4, which may indicate a strong dependence on nitrate assimilation during growth and was used to develop a non-growth media formulation for intrinsic kinetics studies. Photosynthetic intensification factors (PIF) (O2 production by nitrate-limited cyanobacteria in latex coatings/O2 produced by nitrate-limited cell suspensions) in cyanobacteria biocomposites prepared from wet cell pellets concentrated 100- to 300-fold show 7-10 times higher specific reactivity compared to cells in suspension under identical nitrate-limited non-growth conditions. This is the first report of changes of cyanobacteria tolerance to temperature and light intensities after deposition as a thin coating on a porous matrix, which has important implications for gas-phase photobioreactor design using porous composite materials. Cryo-fracture SEM and confocal microscopy images of cell coating distribution on the paper biocomposite suggest that the spatial arrangement of the cells in the coating can affect photoreactivity. This technique could be used to fabricate very stable, multi-organism composite coatings on flexible microfluidic devices in the gas-phase capable of harvesting light in a broader range of wavelengths, to optimize thermotolerant, desiccation tolerant, or halotolerant cyanobacteria that produce O2 with secretion of liquid-fuel precursors synthesized from CO2 .
我们描述了一种乳胶湿法聚结挤出涂层方法,该方法可使沉积在纸张上、水合后置于小型管式光生物反应器气相中的硝酸盐限制的非生长蓝细菌的比光合速率提高多达10倍。这些植物叶状生物复合材料被用于使用太阳模拟器研究蓝细菌菌株对光照和温度的耐受性。我们报告了非生长的聚球藻PCC7002、集胞藻PCC6803、集胞藻PCC6308和鱼腥藻PCC7120的水合气相纸涂层持续500小时的二氧化碳吸收和氧气产生。通过乳胶粘合剂固定在纸张表面的硝酸盐饥饿蓝细菌没有从涂层中生长到本体液体中。聚球藻涂层中的平均二氧化碳消耗速率为5.67 mmol m(-2) h(-1) ,这与文献中报道的拟南芥叶片在类似实验条件下的速率(18 mmol m(-2) h(-1) )非常接近。我们观察到氧气产生与二氧化碳消耗的平均比率(光合商,PQ)在1.3至1.4之间,这可能表明生长过程中对硝酸盐同化有很强的依赖性,并用于开发用于本征动力学研究的非生长培养基配方。由浓缩100至300倍的湿细胞沉淀制备的蓝细菌生物复合材料中的光合强化因子(PIF)(乳胶涂层中硝酸盐限制的蓝细菌产生的氧气/硝酸盐限制的细胞悬浮液产生的氧气)显示,在相同的硝酸盐限制非生长条件下,与悬浮细胞相比,比反应性高7至10倍。这是关于蓝细菌沉积在多孔基质上形成薄涂层后对温度和光强度耐受性变化的首次报告,这对使用多孔复合材料的气相光生物反应器设计具有重要意义。细胞在纸生物复合材料上涂层分布的冷冻断裂扫描电子显微镜和共聚焦显微镜图像表明,涂层中细胞的空间排列会影响光反应性。该技术可用于在气相中的柔性微流控装置上制造非常稳定的多生物体复合涂层,能够在更宽的波长范围内收集光,以优化产生氧气并分泌由二氧化碳合成的液体燃料前体的耐热、耐旱或耐盐蓝细菌。