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Enzymatic pre-treatment of microalgae cells for enhanced extraction of proteins

机译:酶预处理微藻细胞以增强蛋白质提取

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摘要

© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim Crude proteins and pigments were extracted from different microalgae strains, both marine and freshwater. The effectiveness of enzymatic pre-treatment prior to protein extraction was evaluated and compared to conventional techniques, including ultrasonication and high-pressure water extraction. Enzymatic pre-treatment was chosen as it could be carried out at mild shear conditions and does not subject the proteins to high temperatures, as with the ultrasonication approach. Using enzymatic pre-treatment, the extracted proteins yields of all tested microalgae strains were approximately 0.7 mg per mg of dry cell weight. These values were comparable to those achieved using a commercial lytic kit. Ultrasonication was not very ef fective for proteins extraction from Chlorella sp., and the extracted proteins yields did not exceed 0.4 mg per mg of dry cell weight. For other strains, similar yields were achieved by both treatment methods. The time-course effect of enzymatic incubation on the proteins extraction efficiency was more evident using laccase compared to lysozyme, which suggested that the former enzyme has a slower rate of cell disruption. The crude extracted proteins were fractionated using an ion exchange resin and were analyzed by the electrophoresis technique. They were further tested for their antioxidant activity, the highest of which was about 60% from Nannochloropsis sp. The total phenolic contents in the selected strains were also determined, with Chlorella sp. showing the highest content reaching 17 mg/g. Lysozyme was also found to enhance the extraction of pigments, with Chlorella sp. showing the highest pigments contents of 16.02, 4.59 and 5.22 mg/g of chlorophyll a, chlorophyll b and total carotenoids, respectively.
机译:©2016 WILEY-VCH Verlag GmbH&Co. KGaA,Weinheim Weinheim粗蛋白和色素是从海洋和淡水不同微藻菌株中提取的。评估了蛋白质提取之前酶促预处理的有效性,并将其与常规技术(包括超声和高压水提取)进行了比较。选择酶促预处理是因为它可以在温和的剪切条件下进行,并且不会像超声波方法那样使蛋白质经受高温。使用酶预处理,所有测试的微藻菌株的提取蛋白产量约为每毫克干细胞重量0.7毫克。这些值与使用商业裂解试剂盒获得的值相当。超声波对从小球藻中提取蛋白质不是很有效,并且提取的蛋白质产量不超过每毫克干细胞重量0.4毫克。对于其他菌株,两种处理方法均获得了相似的产量。与溶菌酶相比,使用漆酶的酶促孵育对蛋白质提取效率的时程影响更为明显,这表明前者的酶对细胞的破坏速度较慢。使用离子交换树脂对提取的粗蛋白进行分级分离,并通过电泳技术进行分析。进一步测试了它们的抗氧化活性,其中最高的约为Nannochloropsis sp。的60%。还用小球藻(Chlorella sp。)测定了所选菌株中的总酚含量。显示最高含量达到17 mg / g。还发现溶菌酶能增强小球藻的色素提取。分别显示出最高的色素含量,分别为16.02、4.59和5.22 mg / g叶绿素a,叶绿素b和总类胡萝卜素。

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