Analysis of soluble proteins in natural Cordyceps sinensis from different producing areas by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and two-dimensional electrophoresis

Abstract

Background:As one of the bioactive components in Cordyceps sinensis (CS), proteins were rarely used as index components to study the correlation between the protein components and producing areas of natural CS.Objective:Protein components of 26 natural CS samples produced in Qinghai, Tibet, and Sichuan provinces were analyzed and compared to investigate the relationship among 26 different producing areas.Materials and Methods:Proteins from 26 different producing areas were extracted by Tris-HCl buffer with Triton X-100, and separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional electrophoresis (2-DE).Results:The SDS-PAGE results indicated that the number of protein bands and optical density curves of proteins in 26 CS samples was a bit different. However, the 2-DE results showed that the numbers and abundance of protein spots in protein profiles of 26 samples were obviously different and showed certain association with producing areas.Conclusions:Based on the expression values of matched protein spots, 26 batches of CS samples can be divided into two main categories (Tibet and Qinghai) by hierarchical cluster analysis.SUMMARY The number of protein bands and optical density curves of proteins in 26 Cordyceps sinensis samples were a bit different on the sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein profilesNumbers and abundance of protein spots in protein profiles of 26 samples were obvious different on two-dimensional electrophoresis mapsTwenty-six different producing areas of natural Cordyceps sinensis samples were divided into two main categories (Tibet and Qinghai) by Hierarchical cluster analysis based on the values of matched protein spots. Abbreviations Used: SDS-PAGE: Sodium dodecyl sulfate polyacrylamide gel electrophoresis, 2-DE: Two-dimensional electrophoresis, Cordyceps sinensis: CS, TCMs: Traditional Chinese medicines

Identification of Ophiocordyceps sinensis and Its Artificially Cultured Ophiocordyceps Mycelia by Ultra-Performance Liquid Chromatography/Orbitrap Fusion Mass Spectrometry and Chemometrics

“…The fungal protein is a special protein that is differentiated by the type of fungus [40]. One efficient method for proteomic analysis is biological mass spectrometry [39,53,54].…”

“…In recent years, many studies have focused on the fungal proteins. Two dimensional electrophoresis (2-DE) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was used to examine and identify O. sinensis [40,55]. Then, five proteins were identified using MALDI-TOF-TOF/MS.…”

“…The results from the measurements of the aforementioned five samples revealed the following information on the marker peptides: TLLEAIDSIEPPK was only present in wild O. sinensis , AVLSDAITLVR was only detected in fermented O. sinensis mycelium powder, FAELLEK was only observed in fermented Ophiocordyceps mycelium powder, LESVVTSFTK was only present in artificial O. mycelium powder, and VPSSAVLR was only found in O. mortierella mycelium powder. Previous studies revealed the proteins from 26 different producing areas were obviously different in the numbers and abundance of protein spots of protein profiles, and this showed certain association with producing areas [40]. Another report revealed five proteins of O. sinensis were identified in 2-DE, but the specific protein was not reported [55].…”

“…Moreover, these proteins are among the bioactive components in O.S. and, to the best of our knowledge, have rarely been studied and reported [40]. Biological mass spectrometry has been developed as an efficient method for proteomic analysis that exhibits superior mass accuracy and ultra-high resolution, and employs a segmented quadruple mass filter with improved selectivity and ion transmissibility [38,39,41,42,43,44,45,46].…”

Media Optimization for Production of Recombinant Carrier Protein (CRM197) in <i>Escherichia coli</i>

“…Reducing SDS PAGE technique was performed for separation of the expressed protein and analysis as per standard protocol [13]. 12 % tris-glycine gel was used under reducing condition.…”