Acién Fernández Francisco G, Alías Celeste Brindley, García-Malea López María C, Fernández Sevilla José M, Ibáñez González María J, Gómez Rafael Núñez, Molina Grima Emilio
Department of Chemical Engineering, University of Almería, E-04071 Almería, Spain.
Biomol Eng. 2003 Jul;20(4-6):149-62. doi: 10.1016/s1389-0344(03)00041-8.
An integrated process for the indoor production of 13C labeled polyunsaturated fatty acids (PUFAs) from Phaeodactylum tricornutum is presented. The core of the process is a bubble column photobioreactor operating with recirculation of the exhaust gas using a low-pressure compressor. Oxygen accumulation in the system is avoided by bubbling the exhaust gas from the reactor in a sodium sulfite solution before returning to it. To achieve a high 13C enrichment in the biomass obtained, the culture medium is initially stripped of carbon, and labeled 13CO(2) is automatically injected on-demand during operation for pH control and carbon supply. The reactor was operated in both batch and semicontinuous modes. In semicontinuous mode, the reactor was operated at a dilution rate of 0.01 h(-1), resulting in a biomass productivity of 0.1 g l(-1) per day. The elemental analysis of the inlet and outlet flows of the reactor showed that 64.9% of carbon was turned into microalgal biomass, 34.9% remained in the supernatant mainly as inorganic compounds. Only 3.8% of injected carbon was effectively fixed as the target labeled product (EPA). Regarding the isotopic composition of fatty acids, results showed that fatty acids were not labeled in the same proportion, the higher the number of carbons the lower the percentage of 13C. Isotopic composition of EPA ranged from 36.5 to 53.5%, as a function of the methodology used (GC-MS, EA-IRMS or gas chromatography-combustion-isotope ratio mass spectrometry (GC-IRMS)). The low carbon uptake efficiency combined with the high cost of 13CO(2) make necessary to redefine the designed culture system to increase the efficiency of the conversion of 13CO(2) into the target product. Therefore, the possibility of removing 12C from the fresh medium, and recovering and recirculating the inorganic carbon in the supernatant and the organic carbon from the EPA depleted biomass was studied. The inorganic carbon of the fresh medium was removed by acidification and stripping with N(2). The inorganic carbon of the supernatant was recovered also by acidification and subsequent stripping with N(2). The operating conditions of this step were optimized for gas flow rate and type of contactor. A carbon recovery step for the depleted biomass was designed based on the catalytic oxidation to CO(2) using CuO (10 wt.%) as catalyst with an oxygen enriched atmosphere (80% O(2) partial pressure). In this way, the carbon losses reduced an 80.2% and the efficiency of the conversion of carbon in EPA was increased to 19.5%, which is close to the theoretical maximum. Further increase in 13CO(2) use efficiency is only possible by additionally recovering other labeled by-products present in the biomass: proteins, carbohydrates, lipids, and pigments.
本文介绍了一种室内生产来自三角褐指藻的13C标记多不饱和脂肪酸(PUFAs)的综合工艺。该工艺的核心是一个鼓泡塔光生物反应器,通过低压压缩机使废气循环运行。在废气返回反应器之前,将其鼓泡通入亚硫酸钠溶液中,以避免系统中氧气的积累。为了使获得的生物质具有高13C富集度,最初要去除培养基中的碳,并在运行期间根据pH控制和碳供应的需求自动注入标记的13CO₂。反应器以分批和半连续模式运行。在半连续模式下,反应器以0.01 h⁻¹的稀释率运行,生物质生产率为每天0.1 g l⁻¹。对反应器进出口物流的元素分析表明,64.9%的碳转化为微藻生物质,34.9%主要以无机化合物的形式留在上清液中。仅3.8%注入的碳有效地固定为目标标记产物(EPA)。关于脂肪酸的同位素组成,结果表明脂肪酸的标记比例不同,碳数越高,13C的百分比越低。EPA的同位素组成范围为36.5%至53.5%,这取决于所使用的方法(气相色谱 - 质谱法、元素分析仪 - 同位素比率质谱仪或气相色谱 - 燃烧 - 同位素比率质谱仪(GC - IRMS))。低碳吸收效率与13CO₂的高成本使得有必要重新定义设计的培养系统,以提高13CO₂转化为目标产物的效率。因此,研究了从新鲜培养基中去除12C以及回收和循环利用上清液中的无机碳和来自EPA耗尽生物质中的有机碳的可能性。通过酸化并用N₂汽提去除新鲜培养基中的无机碳。上清液中的无机碳也通过酸化并随后用N₂汽提回收。对该步骤的操作条件进行了优化,以确定气体流速和接触器类型。基于使用CuO(10 wt.%)作为催化剂在富氧气氛(80% O₂分压)下催化氧化为CO₂,设计了一个针对耗尽生物质的碳回收步骤。通过这种方式,碳损失减少了80.2%,EPA中碳的转化效率提高到19.5%,接近理论最大值。只有通过额外回收生物质中存在的其他标记副产物:蛋白质、碳水化合物、脂质和色素,才能进一步提高13CO₂的使用效率。