Burdějová Lenka, Dadajová Pavlína, Kudláčková Barbora, Duša Filip
Institute of Analytical Chemistry of the Czech Academy of Sciences, Brno, Czech Republic.
Department of Chemistry, Faculty of Science, Masaryk University, Brno, Czech Republic.
Phytochem Anal. 2025 Mar;36(2):458-466. doi: 10.1002/pca.3486. Epub 2024 Dec 10.
Cyanobacterium Arthrospira platensis (AP) (Nordstedt) Gomont contains high content of phycobiliproteins (PBP), which are an important source for food industry. Methods effectively extracting proteins contained in AP cells are demanded to provide a supply of the material. Water-based extraction methods are advisable due to the high solubility of the PBP.
Extraction techniques such as ultrasound assisted extraction (UAE) and pressurized water extraction (PWE) are popular due to their environmental friendliness, better extraction efficiency, and faster extraction process. In this paper, efficiency of the two methods is compared.
PWE along with UAE is utilized for release of PBP from the AP cells. The extraction parameters including time, temperature, pressure, and ultrasound intensity are tested to obtain the most efficient setup. The methods were evaluated using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and the replicates of PWE extracts were further analyzed by capillary isoelectric focusing with laser-induced fluorescence (cIEF-LIF).
The developed PWE method using higher pressure treatment at lower temperature was significantly faster than UAE methods, and the SDS-PAGE results showed a high content of phycobiliproteins in the extracts. cIEF-LIF analysis showed that the sequential PWE of individual samples was repeatable, and the mild extraction provided a fluorescent profile similar to the commercially available C-phycocyanin standard.
Pressurized water extraction was shown to be an efficient, rapid, and well-automated extraction method for AP proteins in general, including bioactive phycobiliproteins. Obtained results encourage the use of PWE in small-scale analytical applications for primary extraction of proteins.
钝顶节旋藻（AP）（诺德施泰特）戈蒙特含有高含量的藻胆蛋白（PBP），是食品工业的重要原料来源。需要有效的方法来提取AP细胞中所含的蛋白质，以提供该物质的供应。由于PBP的高溶解性，水基提取方法是可取的。
超声辅助提取（UAE）和加压水提取（PWE）等提取技术因其环境友好、提取效率更高和提取过程更快而受到欢迎。本文比较了这两种方法的效率。
采用PWE和UAE从AP细胞中释放PBP。测试提取参数，包括时间、温度、压力和超声强度，以获得最有效的设置。使用十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）对方法进行评估，PWE提取物的重复样品通过毛细管等电聚焦-激光诱导荧光（cIEF-LIF）进一步分析。
所开发的在较低温度下进行更高压力处理的PWE方法明显比UAE方法更快，SDS-PAGE结果显示提取物中藻胆蛋白含量高。cIEF-LIF分析表明，单个样品的连续PWE是可重复的，温和提取提供了与市售C-藻蓝蛋白标准品相似的荧光图谱。
总体而言，加压水提取是一种高效、快速且自动化程度高的提取AP蛋白（包括生物活性藻胆蛋白）的方法。所得结果鼓励在小规模分析应用中使用PWE进行蛋白质的初步提取。
