J Food Bioact

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

Journal website www.isnff-jfb.com

Original Research

Volume 18, June 2022, pages 109-116

Bioactivities of Calocybe indica protein combined with cell-free supernatant of Lactobacilli from a fermented cereal against free radicals and microorganisms

Figure 1. ABTS radical scavenging activity. CIP1: 0.2 mg/ml protein; CIP2: 0.6 mg/ml protein; CFS1: L. lactis CFS; CFS2: L. fermentum CFS; CFS3: L. brevis CFS; BHT: butylated hydroxytoluene (positive control). Error bar is standard error.

Figure 2. DPPH radical scavenging activity. CIP1: 0.2 mg/ml protein; CIP2: 0.6 mg/ml protein; CFS1: L. lactis CFS; CFS2: L. fermentum CFS; CFS3: L. brevis CFS; BHT: butylated hydroxytoluene (positive control). Error bar is standard error

Figure 3. Hydroxy radical scavenging activity. CIP1: 0.2 mg/ml protein; CIP2: 0.6 mg/ml protein; CFS1: L. lactis CFS; CFS2: L. fermentum CFS; CFS3: L. brevis CFS; BHT: butylated hydroxytoluene (positive control). Error bar is standard error

Figure 4. Fe²⁺ chelating activity. CIP1: 0.2 mg/ml protein; CIP2: 0.6 mg/ml protein; CFS1: L. lactis CFS; CFS2: L. fermentum CFS; CFS3: L. brevis CFS; BHT: butylated hydroxytoluene (positive control). Error bar is standard error

**Table 1.** Protein content of *C. indica* and *Lactobacilli* cell-free supernatant
Fraction	Protein concentration (mg/ml)
*Lyophilized protein after dialysis.
C. indica protein (CIP)	7.38 ± 0.00
C. indica protein (CIP)*	0.60 ± 0.01
L. lactis cell-free supernatant (CFS1)	10.58 ± 0.09
L. fermentum cell-free supernatant (CFS2)	9.97 ± 0.14
L. brevis cell-free supernatant (CFS3)	11.89 ± 0.02

Table 1. Protein content of C. indica and Lactobacilli cell-free supernatant

Fraction

Protein concentration (mg/ml)

*Lyophilized protein after dialysis.

C. indica protein (CIP)

7.38 ± 0.00

C. indica protein (CIP)*

0.60 ± 0.01

L. lactis cell-free supernatant (CFS1)

10.58 ± 0.09

L. fermentum cell-free supernatant (CFS2)

9.97 ± 0.14

L. brevis cell-free supernatant (CFS3)

11.89 ± 0.02

**Table 2.** Zones of inhibition (mm) displayed by CIP and CFS of *Lactobacilli* against microorganisms isolated form fruits
Microorganisms	CIP1	CIP2	CFS1	CFS2	CFS3	CIP1 + CFS1	CIP1 + CFS2	CIP1 + CFS3	CIP2 + CFS1	CIP2 + CFS2	CIP2 + CFS3	AMO/KET
^aNo zones of inhibition observed at selected concentration. CIP1: C. indica protein at 0.2 mg/ml, CIP2: C. indica protein at 0.6 mg/ml, CFS1: cell-free supernatant of L. lactis, CFS2: cell-free supernatant of L. fermentum, CFS3: cell-free supernatant of L. brevis; AMO: Amoxicillin (200 µg/ml); KET: ketoconazole (100 µg/ml) against bacteria and fungi, respectively.
Bacillus cereus	5.0 ± 0.0	7.2 ± 0.1	0.0^a	0.0^a	0.0^a	6.6 ± 0.0	8.4 ± 0.1	8.0 ± 0.0	8.1 ± 0.0	10.3 ± 0.8	9.2 ± 0.0	16.0 ± 3.1
Serratia marcescens	5.3 ± 0.2	8.2 ± 0.1	5.7 ± 0.0	0.0^a	5.8 ± 0.3	6.5 ± 0.2	8.0 ± 0.2	8.3 ± 0.2	10.3 ± 1.3	9.5 ± 0.4	9.0 ± 0.1	10.2 ± 1.1
Pseudomonas aeruginosa	0.0^a	0.0^a	0.0^a	0.0^a	6.4 ± 0.0	7.4 ± 0.03	7.0 ± 0.4	7.4 ± 0.3	7.2 ± 0.4	9.2 ± 0.3	9.7 ± 0.1	12.6 ± 1.6
Geotrichum candidum	5.1 ± 0.0	6.5 ± 0.3	5.7 ± 0.0	7.0 ± 0.1	6.4 ± 0.2	7.1 ± 0.2	9.6 ± 0.3	10.2 ± 0.4	9.0 ± 0.2	8.4 ± 04	8.3 ± 0.3	10.8 ± 0.3
Pichia fermentans	5.0 ± 0.1	5.8 ± 0.0	6.0 ± 0.0	0.0^a	0.0^a	8.0 ± 0.2	7.8 ± 0.3	8.5 ± 0.2	10.3 ± 0.3	9.3 ± 0.3	11.2 ± 0.4	15.5 ± 2.0
Aspergillus niger	0.0^a	5.3 ± 0.0	5.1 ± 0.0	6.3 ± 0.2	7.3 ± 0.0	6.8 ± 0.3	8.0 ± 0.03	7.9 ± 0.3	9.7 ± 0.1	11.3 ± 0.3	9.4 ± 0.2	17.9 ± 1.7
Aspergillus flavus	0.0^a	6.7 ± 0.1	0.0^a	5.0 ± 0.0	0.0^a	7.0 ± 0.1	8.1 ± 0.2	10.4 ± 0.1	9.3 ± 0.0	12.1 ± 1.0	10.4 ± 0.4	13.3 ± 0.7
Fusarium solani	0.0^a	0.0^a	6.0 ± 0.0	5.8 ± 0.1	0.0^a	7.3 ± 0.1	7.5 ± 0.1	9.7 ± 0.0	10.6 ± 1.2	10.5 ± 0.7	9.5 ± 0.0	16.1 ± 1.8

Table 2. Zones of inhibition (mm) displayed by CIP and CFS of Lactobacilli against microorganisms isolated form fruits

Microorganisms

CIP1

CIP2

CFS1

CFS2

CFS3

CIP1 + CFS1

CIP1 + CFS2

CIP1 + CFS3

CIP2 + CFS1

CIP2 + CFS2

CIP2 + CFS3

AMO/KET

^aNo zones of inhibition observed at selected concentration. CIP1: C. indica protein at 0.2 mg/ml, CIP2: C. indica protein at 0.6 mg/ml, CFS1: cell-free supernatant of L. lactis, CFS2: cell-free supernatant of L. fermentum, CFS3: cell-free supernatant of L. brevis; AMO: Amoxicillin (200 µg/ml); KET: ketoconazole (100 µg/ml) against bacteria and fungi, respectively.

Bacillus cereus

5.0 ± 0.0

7.2 ± 0.1

0.0^a

6.6 ± 0.0

8.4 ± 0.1

8.0 ± 0.0

8.1 ± 0.0

10.3 ± 0.8

9.2 ± 0.0

16.0 ± 3.1

Serratia marcescens

5.3 ± 0.2

8.2 ± 0.1

5.7 ± 0.0

0.0^a

5.8 ± 0.3

6.5 ± 0.2

8.0 ± 0.2

8.3 ± 0.2

10.3 ± 1.3

9.5 ± 0.4

9.0 ± 0.1

10.2 ± 1.1

Pseudomonas aeruginosa

0.0^a

6.4 ± 0.0

7.4 ± 0.03

7.0 ± 0.4

7.4 ± 0.3

7.2 ± 0.4

9.2 ± 0.3

9.7 ± 0.1

12.6 ± 1.6

Geotrichum candidum

5.1 ± 0.0

6.5 ± 0.3

5.7 ± 0.0

7.0 ± 0.1

6.4 ± 0.2

7.1 ± 0.2

9.6 ± 0.3

10.2 ± 0.4

9.0 ± 0.2

8.4 ± 04

8.3 ± 0.3

10.8 ± 0.3

Pichia fermentans

5.0 ± 0.1

5.8 ± 0.0

6.0 ± 0.0

0.0^a

8.0 ± 0.2

7.8 ± 0.3

8.5 ± 0.2

10.3 ± 0.3

9.3 ± 0.3

11.2 ± 0.4

15.5 ± 2.0

Aspergillus niger

0.0^a

5.3 ± 0.0

5.1 ± 0.0

6.3 ± 0.2

7.3 ± 0.0

6.8 ± 0.3

8.0 ± 0.03

7.9 ± 0.3

9.7 ± 0.1

11.3 ± 0.3

9.4 ± 0.2

17.9 ± 1.7

Aspergillus flavus

0.0^a

6.7 ± 0.1

0.0^a

5.0 ± 0.0

0.0^a

7.0 ± 0.1

8.1 ± 0.2

10.4 ± 0.1

9.3 ± 0.0

12.1 ± 1.0

10.4 ± 0.4

13.3 ± 0.7

Fusarium solani

0.0^a

6.0 ± 0.0

5.8 ± 0.1

0.0^a

7.3 ± 0.1

7.5 ± 0.1

9.7 ± 0.0

10.6 ± 1.2

10.5 ± 0.7

9.5 ± 0.0

16.1 ± 1.8