J Food Bioact

Journal of Food Bioactives, ISSN 2637-8752 print, 2637-8779 online

Journal website www.isnff-jfb.com

Original Research

Volume 20, December 2022, pages 17-28

Flavor compounds, free amino acids, and proteins in Agaricus bisporus mushroom powder

Figure 1. Mushroom powder protein isolation scheme. All extractions were 20 min. 50% NaOH and 1 N HCl were used as needed to adjust extraction pH back to setpoint immediately after mushroom powder addition.

Figure 3. SDS-PAGE characterization of the following samples: A. Molecular weight standard (3–198 kDa); B. Soy protein isolate (1.5% solution, w/w); C. Mycelia (33.3% solution, w/w); D. Mushroom powder 2 (4.6% solution, w/w); E. Mushroom powder 1 (4.6% solution, w/w); F. Alkaline extraction supernatant 1 (18.4% solution, w/w)-pH 10, 20 min, ambient; G. Alkaline extraction supernatant 2 (18.4% solution, w/w)-pH 12, 20 min, ambient; H. Alkaline extraction supernatant 3 (18.4% solution, w/w)-pH 10, 20 min, 50 °C; I. Alkaline extraction supernatant 4 (18.4% solution, w/w)-pH 12, 20 min, 50 °C; and J. Soy protein isolate (1.5% solution, w/w).

**Table 1.** Content (mean ± SD, mg/g dry matter) of five soluble sugars, five organic acids, and five 5′-nucleotides in four samples
	Soy protein isolate	Mycelium	Mushroom powder 2	Mushroom powder 1
Different letters (a–d) within each row indicate significant differences according to one-way ANOVA with Tukey’s HSD (p < 0.05) comparison across four samples.
Glucose	15.78 ± 0.18 b	3.43 ± 0.21 a	62.27 ± 1.25 d	38.75 ± 1.01 c
Fructose	14.46 ± 0.54 d	8.19 ± 0.10 c	4.30 ± 0.50 a	5.58 ± 0.13 b
Sucrose	25.25 ± 0.38 b	9.95 ± 0.03 a	34.83 ± 0.25 c	44.00 ± 0.15 d
Mannitol	1.48 ± 0.04 a	2.89 ± 0.02 b	55.77 ± 1.09 c	69.48 ± 0.18 d
Ribose	1.86 ± 0.05 a	3.64 ± 0.13 b	8.10 ± 0.21 d	6.90 ± 0.22 c
Malic acid	0.38 ± 0.02 b	0.13 ± 0.01 a	1.54 ± 0.09 c	1.91 ± 0.10 d
Acetic acid	0.53 ± 0.03 a	0.57 ± 0.04 a	2.77 ± 0.16 b	3.54 ± 0.23 c
Oxalic acid	0.59 ± 0.02 a	0.75 ± 0.05 a	3.00 ± 0.19 b	4.82 ± 0.13 c
Fumaric acid	0.01 ± 0.00 a	0.00 ± 0.00 a	0.10 ± 0.01 b	0.15 ± 0.01 c
Citric acid	0.28 ± 0.02 a	0.36 ± 0.02 a	1.11 ± 0.08 b	1.86 ± 0.14 c
5′-IMP	0.03 ± 0.00 a	0.02 ± 0.00 a	0.21 ± 0.02 c	0.14 ± 0.00 b
5′-GMP	0.05 ± 0.00 a	0.02 ± 0.00 a	0.75 ± 0.07 b	1.08 ± 0.07 c
5′-AMP	0.17 ± 0.01 b	0.04 ± 0.00 a	0.19 ± 0.01 b	0.36 ± 0.02 c
5′-CMP	0.04 ± 0.00 a	0.02 ± 0.00 a	0.66 ± 0.05 c	0.24 ± 0.01 b
5′-UMP	0.03 ± 0.00 a	0.01 ± 0.00 a	0.29 ± 0.03 b	0.29 ± 0.02 b

Table 1. Content (mean ± SD, mg/g dry matter) of five soluble sugars, five organic acids, and five 5′-nucleotides in four samples

Soy protein isolate

Mycelium

Mushroom powder 2

Mushroom powder 1

Different letters (a–d) within each row indicate significant differences according to one-way ANOVA with Tukey’s HSD (p < 0.05) comparison across four samples.

Glucose

15.78 ± 0.18 b

3.43 ± 0.21 a

62.27 ± 1.25 d

38.75 ± 1.01 c

Fructose

14.46 ± 0.54 d

8.19 ± 0.10 c

4.30 ± 0.50 a

5.58 ± 0.13 b

Sucrose

25.25 ± 0.38 b

9.95 ± 0.03 a

34.83 ± 0.25 c

44.00 ± 0.15 d

Mannitol

1.48 ± 0.04 a

2.89 ± 0.02 b

55.77 ± 1.09 c

69.48 ± 0.18 d

Ribose

1.86 ± 0.05 a

3.64 ± 0.13 b

8.10 ± 0.21 d

6.90 ± 0.22 c

Malic acid

0.38 ± 0.02 b

0.13 ± 0.01 a

1.54 ± 0.09 c

1.91 ± 0.10 d

Acetic acid

0.53 ± 0.03 a

0.57 ± 0.04 a

2.77 ± 0.16 b

3.54 ± 0.23 c

Oxalic acid

0.59 ± 0.02 a

0.75 ± 0.05 a

3.00 ± 0.19 b

4.82 ± 0.13 c

Fumaric acid

0.01 ± 0.00 a

0.00 ± 0.00 a

0.10 ± 0.01 b

0.15 ± 0.01 c

Citric acid

0.28 ± 0.02 a

0.36 ± 0.02 a

1.11 ± 0.08 b

1.86 ± 0.14 c

5′-IMP

0.03 ± 0.00 a

0.02 ± 0.00 a

0.21 ± 0.02 c

0.14 ± 0.00 b

5′-GMP

0.05 ± 0.00 a

0.02 ± 0.00 a

0.75 ± 0.07 b

1.08 ± 0.07 c

5′-AMP

0.17 ± 0.01 b

0.04 ± 0.00 a

0.19 ± 0.01 b

0.36 ± 0.02 c

5′-CMP

0.04 ± 0.00 a

0.02 ± 0.00 a

0.66 ± 0.05 c

0.24 ± 0.01 b

5′-UMP

0.03 ± 0.00 a

0.01 ± 0.00 a

0.29 ± 0.03 b

0.29 ± 0.02 b

**Table 2.** Content (mean ± SD, mg/100g dry matter) of 23 free amino acids in four samples
		Soy protein isolate	Mycelium	Mushroom powder 2	Mushroom powder 1
“–” means not available. Different letters (a–d) within each row indicate significant differences according to one-way ANOVA with Tukey’s HSD (p < 0.05) comparison across four samples.
Umami	Asp	1.82 ± 0.18 a	2.66 ± 0.09 a	271.50 ± 27.95 b	672.05 ± 48.82 c
Umami	Glu	4.90 ± 0.70 a	3.62 ± 0.23 a	1,430.30 ± 104.64 c	1,094.77 ± 124.81 b
Sweet	Ala	5.95 ± 0.19 a	2.71 ± 0.27 a	457.79 ± 18.18 b	450.45 ± 16.93 b
Sweet	Gly	1.43 ± 0.25 a	0.93 ± 0.11 a	97.71 ± 2.09 c	80.97 ± 3.57 b
Sweet	Gln	0.00 ± 0.00 a	5.04 ± 0.47 a	2,070.91 ± 167.63 b	86.62 ± 13.20 a
Sweet	Pro	1.03 ± 0.06 a	1.72 ± 0.27 a	304.62 ± 2.26 c	186.61 ± 3.15 b
Sweet	Ser	0.00 ± 0.00 a	0.00 ± 0.00 a	401.96 ± 43.15 b	455.76 ± 76.09 b
Sweet, E	Thr	0.00 ± 0.00 a	0.49 ± 0.84 a	83.13 ± 7.80 b	135.98 ± 17.17 c
Bitter	Arg	18.17 ± 1.48 a	19.48 ± 0.20 a	18.14 ± 3.01 a	32.86 ± 0.85 b
Bitter, E	His	1.38 ± 1.19 a	0.00 ± 0.00 a	66.59 ± 4.68 c	14.80 ± 1.24 b
Bitter, E	Ile	4.25 ± 0.49 a	0.66 ± 0.08 a	157.74 ± 5.10 b	165.46 ± 9.99 b
Bitter, E	Leu	10.59 ± 1.11 a	1.49 ± 1.10 a	116.00 ± 5.03 c	93.17 ± 5.24 b
Bitter, E	Met	3.57 ± 0.43 b	0.24 ± 0.41 a	2.72 ± 0.36 b	0.00 ± 0.00 a
Bitter, E	Phe	10.93 ± 0.77 a	0.91 ± 0.07 a	153.23 ± 7.05 c	61.61 ± 5.98 b
Bitter	Tyr	4.79 ± 0.34 a	1.52 ± 0.40 a	4.13 ± 0.89 a	54.56 ± 3.21 b
Bitter, E	Val	4.46 ± 0.61 a	1.52 ± 0.27 a	170.27 ± 5.77 c	155.98 ± 3.20 b
Tasteless	Asn	1.79 ± 0.11 a	2.30 ± 0.27 a	306.71 ± 7.01 c	238.92 ± 5.04 b
Tasteless, E	Lys	3.02 ± 0.34 a	1.24 ± 0.08 a	73.46 ± 10.57 c	23.79 ± 1.78 b
Tasteless, E	Trp	5.50 ± 0.32 a	0.00 ± 0.00 a	62.26 ± 10.94 c	26.20 ± 3.27 b
–	Aaa	0.00 ± 0.00 a	0.00 ± 0.00 a	46.78 ± 4.33 b	0.00 ± 0.00 a
–	Aba	0.53 ± 0.03 b	0.03 ± 0.05 a	6.13 ± 0.09 d	4.57 ± 0.21 c
–	Orn	0.84 ± 0.07 a	0.59 ± 0.05 a	196.62 ± 8.36 c	84.66 ± 1.38 b
–	aIle	0.04 ± 0.07 a	0.00 ± 0.00 a	1.50 ± 0.84 b	0.00 ± 0.00 a
	Total	85.01 ± 6.04 a	46.83 ± 2.34 a	6,535.07 ± 53.49 c	3,958.23 ± 100.94 b

Table 2. Content (mean ± SD, mg/100g dry matter) of 23 free amino acids in four samples

Soy protein isolate

Mycelium

Mushroom powder 2

Mushroom powder 1

“–” means not available. Different letters (a–d) within each row indicate significant differences according to one-way ANOVA with Tukey’s HSD (p < 0.05) comparison across four samples.

Umami

Asp

1.82 ± 0.18 a

2.66 ± 0.09 a

271.50 ± 27.95 b

672.05 ± 48.82 c

Umami

Glu

4.90 ± 0.70 a

3.62 ± 0.23 a

1,430.30 ± 104.64 c

1,094.77 ± 124.81 b

Sweet

Ala

5.95 ± 0.19 a

2.71 ± 0.27 a

457.79 ± 18.18 b

450.45 ± 16.93 b

Sweet

Gly

1.43 ± 0.25 a

0.93 ± 0.11 a

97.71 ± 2.09 c

80.97 ± 3.57 b

Sweet

Gln

0.00 ± 0.00 a

5.04 ± 0.47 a

2,070.91 ± 167.63 b

86.62 ± 13.20 a

Sweet

Pro

1.03 ± 0.06 a

1.72 ± 0.27 a

304.62 ± 2.26 c

186.61 ± 3.15 b

Sweet

Ser

0.00 ± 0.00 a

401.96 ± 43.15 b

455.76 ± 76.09 b

Sweet, E

Thr

0.00 ± 0.00 a

0.49 ± 0.84 a

83.13 ± 7.80 b

135.98 ± 17.17 c

Bitter

Arg

18.17 ± 1.48 a

19.48 ± 0.20 a

18.14 ± 3.01 a

32.86 ± 0.85 b

Bitter, E

His

1.38 ± 1.19 a

0.00 ± 0.00 a

66.59 ± 4.68 c

14.80 ± 1.24 b

Bitter, E

Ile

4.25 ± 0.49 a

0.66 ± 0.08 a

157.74 ± 5.10 b

165.46 ± 9.99 b

Bitter, E

Leu

10.59 ± 1.11 a

1.49 ± 1.10 a

116.00 ± 5.03 c

93.17 ± 5.24 b

Bitter, E

Met

3.57 ± 0.43 b

0.24 ± 0.41 a

2.72 ± 0.36 b

0.00 ± 0.00 a

Bitter, E

Phe

10.93 ± 0.77 a

0.91 ± 0.07 a

153.23 ± 7.05 c

61.61 ± 5.98 b

Bitter

Tyr

4.79 ± 0.34 a

1.52 ± 0.40 a

4.13 ± 0.89 a

54.56 ± 3.21 b

Bitter, E

Val

4.46 ± 0.61 a

1.52 ± 0.27 a

170.27 ± 5.77 c

155.98 ± 3.20 b

Tasteless

Asn

1.79 ± 0.11 a

2.30 ± 0.27 a

306.71 ± 7.01 c

238.92 ± 5.04 b

Tasteless, E

Lys

3.02 ± 0.34 a

1.24 ± 0.08 a

73.46 ± 10.57 c

23.79 ± 1.78 b

Tasteless, E

Trp

5.50 ± 0.32 a

0.00 ± 0.00 a

62.26 ± 10.94 c

26.20 ± 3.27 b

–

Aaa

0.00 ± 0.00 a

46.78 ± 4.33 b

0.00 ± 0.00 a

–

Aba

0.53 ± 0.03 b

0.03 ± 0.05 a

6.13 ± 0.09 d

4.57 ± 0.21 c

–

Orn

0.84 ± 0.07 a

0.59 ± 0.05 a

196.62 ± 8.36 c

84.66 ± 1.38 b

–

aIle

0.04 ± 0.07 a

0.00 ± 0.00 a

1.50 ± 0.84 b

0.00 ± 0.00 a

Total

85.01 ± 6.04 a

46.83 ± 2.34 a

6,535.07 ± 53.49 c

3,958.23 ± 100.94 b

**Table 3.** Content (mean ± SD, % dry matter) of the proximate moisture, protein, ash, and fiber, as well as estimated carbohydrate content in 12 samples
	Moisture	Protein	Ash	Fiber	Estimated carbohydrate
“–” means not available. Different letters (a–h) within each column indicate significant differences according to one-way ANOVA with Tukey’s HSD (p < 0.05) comparison across 12 samples. Conversion factor for protein in soy was 6.25, while for the remaining three mushroom samples used 4.38.
Soy protein isolate	6.29 ± 0.14 c	86.56 ± 1.04 f	4.73 ± 0.08 b	3.61 ± 0.34 b	−1.18
Mycelium	2.97 ± 0.15 a	5.34 ± 0.18 a	1.64 ± 0.14 a	2.67 ± 0.20 ab	87.37
Mushroom powder 2	6.58 ± 0.18 d	19.90 ± 0.05 d	12.02 ± 0.12 de	18.21 ± 0.32 c	43.30
Mushroom powder 1	5.59 ± 0.14 b	23.88 ± 0.10 e	11.32 ± 0.09 cde	21.57 ± 1.08 d	37.64
Supernatant 1	–	18.34 ± 0.30 c	30.61 ± 1.00 g	1.84 ± 0.12 a	49.22
Supernatant 2	–	18.25 ± 0.02 c	47.98 ± 0.59 h	2.58 ± 0.21 ab	31.19
Supernatant 3	–	18.22 ± 0.29 c	21.49 ± 1.13 f	2.69 ± 0.16 ab	57.61
Supernatant 4	–	18.49 ± 0.16 c	47.26 ± 2.67 h	2.69 ± 0.31 ab	31.56
Pellet 1	–	17.86 ± 0.09 c	9.97 ± 0.02 cd	44.76 ± 1.10 f	27.41
Pellet 2	–	15.75 ± 0.15 b	13.81 ± 0.17 e	42.18 ± 0.66 e	28.26
Pellet 3	–	17.35 ± 0.07 c	9.22 ± 0.06 c	46.81 ± 0.76 g	26.62
Pellet 4	–	15.88 ± 0.13 b	13.05 ± 0.08 e	43.97 ± 0.75 f	27.11

Table 3. Content (mean ± SD, % dry matter) of the proximate moisture, protein, ash, and fiber, as well as estimated carbohydrate content in 12 samples

Moisture

Protein

Ash

Fiber

Estimated carbohydrate

“–” means not available. Different letters (a–h) within each column indicate significant differences according to one-way ANOVA with Tukey’s HSD (p < 0.05) comparison across 12 samples. Conversion factor for protein in soy was 6.25, while for the remaining three mushroom samples used 4.38.

Soy protein isolate

6.29 ± 0.14 c

86.56 ± 1.04 f

4.73 ± 0.08 b

3.61 ± 0.34 b

−1.18

Mycelium

2.97 ± 0.15 a

5.34 ± 0.18 a

1.64 ± 0.14 a

2.67 ± 0.20 ab

87.37

Mushroom powder 2

6.58 ± 0.18 d

19.90 ± 0.05 d

12.02 ± 0.12 de

18.21 ± 0.32 c

43.30

Mushroom powder 1

5.59 ± 0.14 b

23.88 ± 0.10 e

11.32 ± 0.09 cde

21.57 ± 1.08 d

37.64

Supernatant 1

–

18.34 ± 0.30 c

30.61 ± 1.00 g

1.84 ± 0.12 a

49.22

Supernatant 2

–

18.25 ± 0.02 c

47.98 ± 0.59 h

2.58 ± 0.21 ab

31.19

Supernatant 3

–

18.22 ± 0.29 c

21.49 ± 1.13 f

2.69 ± 0.16 ab

57.61

Supernatant 4

–

18.49 ± 0.16 c

47.26 ± 2.67 h

2.69 ± 0.31 ab

31.56

Pellet 1

–

17.86 ± 0.09 c

9.97 ± 0.02 cd

44.76 ± 1.10 f

27.41

Pellet 2

–

15.75 ± 0.15 b

13.81 ± 0.17 e

42.18 ± 0.66 e

28.26

Pellet 3

–

17.35 ± 0.07 c

9.22 ± 0.06 c

46.81 ± 0.76 g

26.62

Pellet 4

–

15.88 ± 0.13 b

13.05 ± 0.08 e

43.97 ± 0.75 f

27.11

**Table 4.** Content (mean ± SD, %) of the soluble solids and soluble protein for 12 samples
	Soluble solids	Soluble protein
Different letters (a–h) within each column indicate significant differences according to one-way ANOVA with Tukey’s HSD (p < 0.05) comparison across 12 samples.
Soy protein isolate	31.28 ± 0.08 d	5.61 ± 0.63 d
Mycelia	4.95 ± 0.36 a	0.10 ± 0.01 a
Mushroom powder 2	67.83 ± 1.13 f	3.61 ± 0.01 c
Mushroom powder 1	63.08 ± 2.32 e	3.65 ± 0.02 c
Supernatant 1	72.68 ± 0.26 h	0.59 ± 0.02 b
Supernatant 2	71.21 ± 0.26 gh	0.54 ± 0.03 b
Supernatant 3	70.06 ± 0.51 g	0.45 ± 0.01 b
Supernatant 4	68.10 ± 0.68 f	0.48 ± 0.05 b
Pellet 1	25.73 ± 0.00 b	0.23 ± 0.00 a
Pellet 2	29.65 ± 0.00 cd	0.20 ± 0.02 a
Pellet 3	26.21 ± 0.00 b	0.19 ± 0.01 a
Pellet 4	28.88 ± 0.00 c	0.17 ± 0.02 a

Table 4. Content (mean ± SD, %) of the soluble solids and soluble protein for 12 samples

Soluble solids

Soluble protein

Different letters (a–h) within each column indicate significant differences according to one-way ANOVA with Tukey’s HSD (p < 0.05) comparison across 12 samples.

Soy protein isolate

31.28 ± 0.08 d

5.61 ± 0.63 d

Mycelia

4.95 ± 0.36 a

0.10 ± 0.01 a

Mushroom powder 2

67.83 ± 1.13 f

3.61 ± 0.01 c

Mushroom powder 1

63.08 ± 2.32 e

3.65 ± 0.02 c

Supernatant 1

72.68 ± 0.26 h

0.59 ± 0.02 b

Supernatant 2

71.21 ± 0.26 gh

0.54 ± 0.03 b

Supernatant 3

70.06 ± 0.51 g

0.45 ± 0.01 b

Supernatant 4

68.10 ± 0.68 f

0.48 ± 0.05 b

Pellet 1

25.73 ± 0.00 b

0.23 ± 0.00 a

Pellet 2

29.65 ± 0.00 cd

0.20 ± 0.02 a

Pellet 3

26.21 ± 0.00 b

0.19 ± 0.01 a

Pellet 4

28.88 ± 0.00 c

0.17 ± 0.02 a