Optimization of Fermentation Conditions of Antagonistic Actinomycetes FS-4

doi:10.3969/j.issn.1000-2561.2020.02.018

Abstract

Abstract:

The screening of antagonistic bacteria from soil microorganisms in the areas with banana Fusarium wilt is of great practical significance for controlling this disease. In this report, a strain, designated FS-4, was isolated from healthy banana rhizosphere soil in the area affected by Fusarium wilt. Based on Gause’s No. 1 synthetic medium, the optimization of medium, single factor experiments and response surface experiments were conducted to maximize the production of the antibacterial substances of actinomycete FS-4. The best fermentation medium and fermentation conditions were as follows: sucrose 2.4%, peptone 0.5%, K₂HPO₄0.05%, NaCl 0.05%, MgSO₄0.05%, fermentation temperature 28 ℃, initial pH 7. Under the conditions, the inhibition zone diameter of actinomycete FS-4 fermentation filtrate reached 27.1 mm on the test plates of Bacillus subtilis after 62 h incubation

Key words: actinomycetes FS-4, fermentation conditions, response surface method

CLC Number:

Q949.748.5

DUAN Yajie,MEI Zhigang,SUN Dequan,LI Weiming,PANG Zhencai,HU Huigang. Optimization of Fermentation Conditions of Antagonistic Actinomycetes FS-4[J]. Chinese Journal of Tropical Crops, 2020, 41(2): 339-345.

Figures/Tables 14

Tab. 1 Box-Behnken experimental for factor and level

因素Factor	水平Level
因素Factor	-1	0	1
A蛋白胨/%	0.3	0.4	0.5
B蔗糖/%	1	2	3
C时间/h	24	48	72

Fig. 1 Effect of carbon sources on activity of antimicrobial substance produced by FS-4 Difference lowercase letters represent 5% significant level.

Fig. 2 Effect of sucrose concentration on activity of antimicrobial substance produced by FS-4 Difference lowercase letters represent 5% significant level.

Fig. 3 Effect of nitrogen sources on activity of antimicrobial substance produced by FS-4 Different lowercase letters represent 5% significant level.

Fig. 4 Effect of nitrogen sources concentration on activity of antimicrobial substance produced by FS-4 Difference lowercase letters represent 5% significant level.

Fig. 5 Effect of sodium chloride, dipotassium hydrogen phosphate and magnesium sulfate concentration on activity of antimicrobial substance produced by fS-4 Difference lowercase letters represent 5% significant level.

Fig.6 Effect of pH on activity of antimicrobial substance produced by FS-4 Difference lowercase letters represent 5% significant level.

Fig. 7 Effect of temperature on activity of antimicrobial substance produced by FS-4 Difference lowercase letters represent 5% significant level.

Fig. 8 Effect of fermentation time on activity of antimicrobial substance produced by FS-4 Difference lowercase letters represent 5% significant level.

Tab. 2 Response surface design experimental results

实验号 Experiment number	A	B	C	抑菌圈直径 Inhibition zone diameter/mm
1	0	0	0	25.4
2	1	0	-1	21.9
3	0	1	-1	20.9
4	-1	-1	0	21.4
5	0	0	0	24.9
6	-1	1	0	22.9
7	-1	0	1	21.4
8	0	0	0	25.4
9	1	-1	0	26.4
10	0	0	0	25.9
11	0	1	1	25.4
12	1	0	1	26.4
13	0	-1	1	24.4
14	1	1	0	26.9
15	0	-1	-1	21.4
16	-1	0	-1	20.4
17	0	0	0	26.7

Tab. 3 ANOVA for model analysis of variance

方差来源 Sources of variation	平方和 Quadratic sum	自由度 Freedom	均方 Mean square	F值 F value	P	显著性 Conspicuousness
模型	82.73	9	9.19	18.85	0.0004	**
A蛋白胨	30.03	1	30.03	61.57	0.0001	**
B蔗糖	0.78	1	0.78	1.60	0.2462
C时间	21.13	1	21.13	43.31	0.0003	**
AB	0.25	1	0.25	0.51	0.4972
AC	3.06	1	3.06	6.28	0.0407	*
BC	0.56	1	0.56	1.15	0.3185
A²	3.26	1	3.26	6.68	0.0362	*
B²	0.61	1	0.61	1.25	0.3011
C²	21.41	1	21.41	43.89	0.0003	**
残差	3.41	7	0.49
失拟项	1.56	3	0.52	1.12	0.4386
纯误差	1.85	4	0.46
总和	86.15	16

Fig. 9 Effect of protein and sucrose content on activity of antimicrobial substance produced by FS-4

Fig. 10 Effect of protein content and time on activity of antimicrobial substance produced by FS-4

Fig. 11 Effect of sucrose content and time on activity of antimicrobial substance produced by FS-4

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