Abstract
ABSTRACTAuricularia auricula is one of the most popular edible mushrooms in China. Limited information is available regarding genes and proteins related to A. auricula development from mycelium to mature fruiting body and response to environmental stimuli. With the development of proteomics, this technology platform for addressing this problem is available. A critical step of a good proteome is protein extraction which can gives the best quality and a wide coverage of total proteins. To establish an effective protein extraction protocol suitable for proteomics analysis in mycelium and fruiting body of A. auricula, four previously reported protein extraction protocols including direct extraction, trichloroacetic acid (TCA) precipitation, TCA/acetone precipitation and phenol-based extraction, were compared based on the protein yield and 2-DE patterns. The efficiency of the four methods was evaluated by comparing protein yield and 2-DE patterns. Of the four methods, both TCA precipitation and phenol-based extraction method for A. auricula mycelia protein extraction yielded the good protein separation pattern and higher number of protein spots in 2-DE analysis. Furthermore, phenol-based extraction is superior to the other tested methods for A. auricula fruiting body proteome analysis .
Highlights
Auricularia auricula forms mutualistic symbioses with many tree species, commonly known as tree-ear, which is an edible mushroom found worldwide
Four different protein extraction methods were evaluated with their consistency and reproducibility in extracting total protein samples from mycelium and fruiting body of A. auricula
There was no significant difference in the protein yield extracted via direct extraction method (Method 1), trichloroacetic acid (TCA) precipitation method (Method 2) and TCA/acetone precipitation method (Method 3) from mycelium and fruiting body (Table 1)
Summary
Auricularia auricula forms mutualistic symbioses with many tree species, commonly known as tree-ear, which is an edible mushroom found worldwide. Several studies have reported diverse biological activities of this mushroom, including anti-tumor and anti-coagulant activity owing to its enrichment of polysaccharide and polyphenolic compounds.[1,2,3] The consumption of A. auricula may potentially contribute to the decreased risk of hypercholesterolemia and improve heart function.[4,5] Much attention has been paid to the bioactive compounds isolation, identification, and characterization in A. auricula, due to their numerous health beneficial effects.[4,6,7,8,9] With the development of modern chromatographic and spectrometric techniques, bioactive compounds in A. auricula has been analyzed easier than before[10,11,12,13], but the regulation of this metabolic process is poorly understood and will require targeted, well-designed and appropriately research to gain more information.
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