Effective protein extraction from mycelium and fruiting body of Auricularia auricula for proteomics studies

Figures & data

Figure 1. Graphical representative of four protein extraction methods.

Table 1. Protein yields obtained from mycelium and fruiting body of A. auricula using four different extraction methods.

Extraction method	Protein yield (mg/g)
	Mycelium	Fruiting body
Method 1	5.21 ± 0.08^a	0.27 ± 0.01^a
Method 2	4.85 ± 0.25^a	0.24 ± 0.01^a
Method 3	5.07 ± 0.24^a	0.24 ± 0.01^a
Method 4	3.89 ± 0.28^b	0.20 ± 0.01^b

Data expressed as mean ± SD (n = 3). Different letters in the same row within each fraction indicate significant difference between mean values (One-way ANOVA followed by SPSS Inc test, p < 0.05).

Figure 2. SDS-PAGE seperation of proteins from mycelium (A) and fruiting body (B) using different protein extraction methods, including direct extraction (Method 1), TCA precipitation extraction (Method 2), TCA/acetone precipitation extraction (Method 3), and phenol-based extraction (Method 4). Totally, 80 μg proteins are resolved using 12.5% polyacrylamide gel and visualized with CBB.

Table 2. Total numbers of detectable spots from mycelium and fruiting body based on different protein extraction methods.

Extraction method	Total numbers of detectable spots
	Mycelium	Fruiting body
Method 1	144 ± 14	-
Method 2	1365 ± 55	1035 ± 33
Method 4	1380 ± 30	1038 ± 127
Method 2*	-	759 ± 91
Method 4*	-	833 ± 45

Method 2* and Method 4* were Method 2 and Method 4 in combination with protein purification procedure.

Data expressed as mean ± SD (n = 3).

Figure 3. 2-DE analysis of extracted proteins from mycelium tissues. Equal amounts (1300 μg) of proteins were loaded on 24 cm pH 3–10 IPG strips. (A) direct extraction method, (B) TCA precipitation extraction method, (C) phenol-based extraction method.

Figure 4. 2-DE analysis of extracted proteins from fruiting body. Equal amounts (1300 μg) of proteins were loaded on 24 cm pH 3–10 IPG strips. (A) TCA precipitation extraction method, (B) phenol/ammonium acetate extraction method, (C) TCA precipitation extraction in combination with protein purification (D) phenol-based extraction in combination with protein purification.