Optimization of Fermentation Culture Conditions for Macrolactins Yield of Bacillus siamensis Using Response Surface Methodology

doi:10.13560/j.cnki.biotech.bull.1985.2024-0040

Abstract

Abstract:

【Objective】Macrolactins（MLNs）are a class of 24-membered macrolides formed by cyclization of polyketones, which have a variety of biological activities such as antibacterial, antifungal, antiviral, and anti-tumor. However, due to the low yield of macrolides in traditional fermentation, its development and application are limited. Therefore, this article aims to increase the yield of MLNs synthesized by Bacillus siamensis K53, in order to provide scientific basis for the mass production of MLNs.【Method】Based on previous research, single factor experiments was used to study the effects of carbon, nitrogen and amino acids on the MLNs yield of strain K53 and to determine the key nutritional factors. Response surface analysis was used to optimize the key nutritional factors for obtaining the optimal parameters in fermenting MLNs.【Result】In the single factor experiment, glucose was the optimal carbon source in the fermentation medium of strain K53. The yeast extract and (NH₄)₂SO₄ were the optimal organic and inorganic nitrogen sources, respectively. The addition of histidine and valine increased the yield of MLNs. In response surface optimization, the optimal fermentation medium for strain K53 was 22.0 g/L glucose, 3.0 g/L (NH₄)₂SO₄, 1.0 g/L yeast extract, 6.1 g/L valine, and 5.9 g/L histidine. The valine and histidine were precursors for the synthesis of MLNs. The MLNs yield in this medium reached 5.37×10² μg/mL, which was very close to the maximum value（5.46×10² μg/mL）predicted by theory, and 4.10 times higher than that in the non-optimized medium（1.31×10² μg/mL）.【Conclusion】The optimized medium formula by response surface methodology is conducive to the MLNs yield of the strain K53，and the results provide theoretical bases for the consequent development and application of the strain K53.

Key words: macrolactins, Bacillus siamensis, fermentation medium, precursor substance, optimization via response surface

ZHANG Meng-fei, YU Lian, LI Fei, LI Zhe, SU Xin-ying, LAN Cai-bi, ZHU Hu, QIN Ying. Optimization of Fermentation Culture Conditions for Macrolactins Yield of Bacillus siamensis Using Response Surface Methodology[J]. Biotechnology Bulletin, 2024, 40(8): 299-308.

Figures/Tables 9

References 37

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因素Factor/（g·L^-1）	低水平Low level（-1）	高水平High level（1）
葡萄糖（X₁）	10	20
酵母提取物（X₂）	0.6	1.0
(NH₄)₂SO₄（X₃）	1.0	3.0
缬氨酸（X₄）	2.0	6.0
组氨酸（X₅）	2.0	6.0

因素Factor/（g·L^-1）	低水平Low level（-1）	高水平High level（1）
葡萄糖（X₁）	10	20
酵母提取物（X₂）	0.6	1.0
(NH₄)₂SO₄（X₃）	1.0	3.0
缬氨酸（X₄）	2.0	6.0
组氨酸（X₅）	2.0	6.0

水平 Level	葡萄糖Glucose X_1/（g·L^-1）	酵母提取物Yeast extract X₂/（g·L^-1）	缬氨酸Valine X₄/（g·L^-1）	组氨酸Histidine X₅/（g·L^-1）
-1	15	0.8	5.0	5.0
0	20	1.0	6.0	6.0
1	25	1.2	7.0	7.0

水平 Level	葡萄糖Glucose X_1/（g·L^-1）	酵母提取物Yeast extract X₂/（g·L^-1）	缬氨酸Valine X₄/（g·L^-1）	组氨酸Histidine X₅/（g·L^-1）
-1	15	0.8	5.0	5.0
0	20	1.0	6.0	6.0
1	25	1.2	7.0	7.0

来源Source	自由度Freedom	离均差平方和Sum of squares of deviation from mean	均方差Standard deviation	F值 F value	P值P value
模型	5	0.170	0.034	15.76	0.002 1
X₁	1	0.014	0.014	6.28	0.046 2
X₂	1	0.089	0.089	40.88	0.000 7
X₃	1	0.001	0.001	0.64	0.453 0
X₄	1	0.047	0.047	21.65	0.003 5
X₅	1	0.021	0.021	9.37	0.022 2
残差	6	0.013	0.002
总离差	11	0.190