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微藻(绿藻门)中蛋白质的比色测定:研磨与SDS处理联合用于总蛋白提取

Colorimetric protein determination in microalgae (Chlorophyta): association of milling and SDS treatment for total protein extraction.

作者信息

Mota Maria Fernanda S, Souza Marcella F, Bon Elba P S, Rodrigues Marcoaurelio A, Freitas Suely Pereira

机构信息

Bioethanol Laboratory, Federal University of Rio de Janeiro, Av. Pedro Calmon, s/n, 21941-596, Rio de Janeiro, RJ, Brazil.

Laboratory of Microalgae Biotechnology and Photosynthesis, Federal University of Rio de Janeiro, Av. Athos da Silveira Ramos, 149, bl A, lab 532, 21941-909, Rio de Janeiro, RJ, Brazil.

出版信息

J Phycol. 2018 Aug;54(4):577-580. doi: 10.1111/jpy.12754. Epub 2018 Jun 19.

Abstract

The use of colorimetric methods for protein quantification in microalgae is hindered by their elevated amounts of membrane-embedded intracellular proteins. In this work, the protein content of three species of microalgae was determined by the Lowry method after the cells were dried, ball-milled, and treated with the detergent sodium dodecyl sulfate (SDS). Results demonstrated that the association of milling and SDS treatment resulted in a 3- to 7-fold increase in protein quantification. Milling promoted microalgal disaggregation and cell wall disruption enabling access of the SDS detergent to the microalgal intracellular membrane proteins and their efficient solubilization and quantification.

摘要

比色法用于微藻蛋白质定量时,会受到其大量膜嵌入细胞内蛋白质的阻碍。在这项工作中,三种微藻细胞经干燥、球磨并用去污剂十二烷基硫酸钠(SDS)处理后,采用洛瑞法测定其蛋白质含量。结果表明,球磨和SDS处理相结合使蛋白质定量增加了3至7倍。球磨促进了微藻的解聚和细胞壁的破坏,使SDS去污剂能够接触到微藻细胞内膜蛋白,并实现其有效溶解和定量。

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