Evaluation of sample extraction methods for proteomics analysis of green algae Chlorella vulgaris

Abstract

Many protein extraction methods have been developed for plant proteome analysis but information is limited on the optimal protein extraction method from algae species. This study evaluated four protein extraction methods, i.e. direct lysis buffer method, TCA-acetone method, phenol method, and phenol/TCA-acetone method, using green algae Chlorella vulgaris for proteome analysis. The data presented showed that phenol/TCA-acetone method was superior to the other three tested methods with regards to shotgun proteomics. Proteins identified using shotgun proteomics were validated using sequential window acquisition of all theoretical fragment-ion spectra (SWATH) technique. Additionally, SWATH provides protein quantitation information from different methods and protein abundance using different protein extraction methods was evaluated. These results highlight the importance of green algae protein extraction method for subsequent MS analysis and identification.

Comparative proteomic analysis reveals the regulatory network of theveAgene during asexual and sexual spore development ofAspergillus cristatus

“…Protein was extracted from the spores according to a protocol described previously [ 20 ]. Briefly, spores of the WT, agamotype, and teleomorph strains were collected from three biological cultures grown on low osmotic pressure solid medium, and washed in a phosphate buffer solution (pH 7.2).…”

Extracting protein from microalgae (Tetraselmis chuii) for proteome analysis

“…Nonetheless, although protein extraction efficiency is an important parameter it is not correlated with protein integrity an important factor for bioactivity and proteomics analysis. Indeed, the reported microalgae protein extraction methods are for analytical and functional purposes, and few have focused on the proteome [ 12 , 13 , [15] , [16] , [17] , [22] , [23] , [24] , [25] , [26] ]. In proteomics studies it is essential to use protein extracts that give representative and consistent, high-quality results.…”

Recent Advances in Microalgae Peptides: Cardiovascular Health Benefits and Analysis

“…Therefore, the unbiased nature of data-independent acquisition makes it the best technique for discovery proteomics. This new methodology has been recently applied as a different method for shotgun proteomics to identify proteins of C. vulgaris in order to select the best extraction protocols . The ability to comprehensively identify peptides in very complex matrices over a large dynamic range and in an extremely reproducible mode opens up a world of application possibilities to bioactive peptide analysis by DIA.…”