Papachristou Ioannis, Nazarova Natalja, Wüstner Rüdiger, Lina Robin, Frey Wolfgang, Silve Aude
Institute for Pulsed Power and Microwave Technology (IHM), Karlsruhe Institute of Technology (KIT), Eggenstein-Leopoldshafen, Germany.
Biotechnol Biofuels Bioprod. 2025 Jan 28;18(1):12. doi: 10.1186/s13068-025-02608-7.
The gradual extrusion of water-soluble intracellular components (such as proteins) from microalgae after pulsed electric field (PEF) treatment is a well-documented phenomenon. This could be utilized in biorefinery applications with lipid extraction taking place after such an 'incubation' period, i.e., a post-PEF-treatment step during which the biomass is left undisturbed before any further processing. The goal of this work was to further explore how this incubation could improve lipid extraction.
Experiments were conducted on wet, freshly harvested Auxenochlorella protothecoides, treated with 0.25 or 1.5 MJ/kg and incubated for 24 h. Lipid extraction took place with a monophasic ethanol:hexane:water, 1:0.41:0.04 vol/vol/vol mixture with a 75.6 mL solvent per 1 g of dry biomass ratio. The kinetics of the extraction were studied with samples taken between 10 and 1080 min from fresh and incubated biomass. The yields at 10 min were significantly increased with incubation compared to without (31.2% dry weight compared to 1.81%, respectively). The experimental data were fitted with the Patricelli model where extraction occurs in two steps, a rapid washing of immediate available lipids and a slower diffusion one. During Nile-Red staining of microalgae and microscopy imaging, a shift of emission from both GFP and RFP channels to mostly RFP was observed indicating an increase in the polarity of the environment of Nile-Red. These led to an adaption of a biphasic ethanol:hexane:water 1:6:0.4 vol/vol/vol solvent with 37 mL solvent per 1 g of dry biomass ratio which while ineffective on fresh biomass, achieved a 27% dry weight yield from incubated microalgae. The extraction efficiency in the biphasic route was lower compared to the monophasic (i.e., 69% and 95%, respectively). It was compensated however, by the significant solvent reduction (37 mL to 75.6 mL respectively), in particular the ethanol minimization. For the extraction of 1 L lipids, it was estimated that the energy consumption ratio for the biphasic process was 1.6 compared to 9.9 for monophasic, making clearly the most preferential one.
This biphasic approach significantly reduces solvent consumption and the respective energy requirement for solvent recovery. Incubation thus could majorly improve the commercialization prospects of the process.
脉冲电场(PEF)处理后,微藻中水溶性细胞内成分(如蛋白质)会逐渐挤出,这是一个有充分记录的现象。这可用于生物炼制应用,即在这样一个“孵育”期后进行脂质提取,即PEF处理后的步骤,在此期间生物质在进一步加工之前保持不受干扰。这项工作的目标是进一步探索这种孵育如何能提高脂质提取率。
对新鲜收获的湿的原壳小球藻进行实验,用0.25或1.5兆焦/千克处理,并孵育24小时。脂质提取使用单相乙醇:己烷:水,1:0.41:0.04体积/体积/体积的混合物,每1克干生物质使用75.6毫升溶剂。从新鲜和孵育后的生物质中在10至1080分钟之间取样研究提取动力学。与未孵育相比,孵育后10分钟时的产量显著增加(分别为干重的31.2%和1.81%)。实验数据用Patricelli模型拟合,其中提取分两步进行,快速洗涤即时可用的脂质,以及较慢的扩散步骤。在微藻的尼罗红染色和显微镜成像过程中,观察到绿色荧光蛋白(GFP)和红色荧光蛋白(RFP)通道的发射都向主要是RFP转变,表明尼罗红环境的极性增加。这导致采用双相乙醇:己烷:水1:6:0.4体积/体积/体积的溶剂,每1克干生物质使用37毫升溶剂,虽然对新鲜生物质无效,但从孵育后的微藻中获得了27%的干重产量。双相途径的提取效率低于单相途径(分别为69%和95%)。然而,通过显著减少溶剂用量(分别从75.6毫升减少到37毫升),特别是乙醇用量的最小化,得到了补偿。对于提取1升脂质,估计双相过程的能量消耗比为1.6,而单相过程为9.9,显然双相过程是最优先选择的。
这种双相方法显著降低了溶剂消耗以及溶剂回收所需的相应能量。因此,孵育可以极大地改善该工艺的商业化前景。