海洋真菌Meira nashicola ZJ353的固态发酵及其代谢产物的分离与鉴定

doi:10.19586/j.2095-2341.2025.0072

生物技术进展 ›› 2025, Vol. 15 ›› Issue (6): 1040-1049.DOI: 10.19586/j.2095-2341.2025.0072

• 研究论文 • 上一篇

海洋真菌Meira nashicola ZJ353的固态发酵及其代谢产物的分离与鉴定

曹静雯¹(), 包艳春¹, 毛远朝¹, 杨馨源¹, 崔海栋², 侯熙彦¹, 金黎明¹()

^1.大连民族大学，生物技术与资源利用教育部重点实验室，辽宁大连 116600
^2.青岛卫辽医用生物材料有限公司，山东青岛 266109

收稿日期:2025-06-30 接受日期:2025-08-21 出版日期:2025-11-25 发布日期:2026-01-04
通讯作者: 金黎明
作者简介:曹静雯 E-mail:1392361708@qq.com；
基金资助:
辽宁省自然科学基金计划面上项目(2024-MS-019);辽宁省科学技术厅应用基础研究计划(2022020332-JH2);大连民族大学大学生创新创业训练计划项目

Solid-state Fermentation of Marine Fungus Meira nashicola ZJ353 and the Isolation and Identification of its Metabolites

Jingwen CAO¹(), Yanchun BAO¹, Yuanchao MAO¹, Xinyuan YANG¹, Haidong CUI², Xiyan HOU¹, Liming JIN¹()

^1.Key Laboratory of Biotechnology and Bioresources Utilization of Ministry of Education，Dalian Minzu University，Liaoning Dalian 116600，China
^2.Qingdao ValueValue Medical Biomaterial Co. ，Ltd. ，Shandong Qingdao 266109，China

Received:2025-06-30 Accepted:2025-08-21 Online:2025-11-25 Published:2026-01-04
Contact: Liming JIN

摘要/Abstract

摘要：

从海洋中开发新型抗生素已成为近年的研究热点。以分离自北冰洋海泥中的21株真菌菌株为研究对象，通过琼脂扩散法筛选对金黄色葡萄球菌具有抑制作用的3株真菌，发现编号ZJ353的菌株抗菌活性最佳。经过形态学鉴定和26S rDNA菌种鉴定，命名为Meira nashicola ZJ353。对其固态发酵条件进行优化，并对代谢产物进行分离鉴定，确定了最佳发酵条件：大米培养基、培养基初始加水量为90%、发酵时间为11 d、发酵温度为30 ℃。对M. nashicola ZJ353进行扩大培养，乙酸乙酯超声浸提其发酵产物，通过硅胶柱层析和高效液相色谱分离出一个化合物单体。结合红外光谱、精确质量飞行时间质谱液质联用技术、¹H NMR和¹³C NMR对化合物的结构进行分析，鉴定出化合物的分子式为C₁₆H₂₂O₄。研究结果为海洋微生物源抗菌物质开发提供了一定的理论基础。

关键词: 海洋真菌, 发酵条件优化, 代谢产物, 分离, 鉴定

Abstract:

In recent years， the development of new antibiotics from the ocean has become a research hotspot. Using 21 fungi strains isolated from arctic sea mud as the research objects， three strains of fungi with inhibitory effects on Staphylococcus aureus were screened out by the agar diffusion method， among which the strain numbered ZJ353 exhibited the best antibacterial activity. Through morphological identification and 26S rDNA species identification， it was named Meira nashicola ZJ353. The solid-state fermentation conditions were optimized， and the metabolites were isolated and identified to determine the optimal fermentation conditions： rice medium， initial water addition of the medium at 90%， fermentation time of 11 days， and fermentation temperature of 30 ℃. Then， the M. nashicola ZJ353 was expanded for cultivation， and its fermentation products were extracted by ultrasonic treatment with ethyl acetate， a compound monomer was isolated through column chromatography on silica gel and HPLC. Using infrared spectroscopy， precise mass time-of-flight liquid chromatography-mass spectrometry， ¹H NMR and ¹³C NMR spectra， the structure of the compound was analyzed， and the molecular formula of the compound was determined to be C₁₆H₂₂O₄. The results provides a certain theoretical basis for the development of antibacterial substances from marine microorganisms.

Key words: marine fungi, fermentation conditions optimization, metabolic products, separation, identification

中图分类号:

Q935

曹静雯, 包艳春, 毛远朝, 杨馨源, 崔海栋, 侯熙彦, 金黎明. 海洋真菌Meira nashicola ZJ353的固态发酵及其代谢产物的分离与鉴定[J]. 生物技术进展, 2025, 15(6): 1040-1049.

Jingwen CAO, Yanchun BAO, Yuanchao MAO, Xinyuan YANG, Haidong CUI, Xiyan HOU, Liming JIN. Solid-state Fermentation of Marine Fungus Meira nashicola ZJ353 and the Isolation and Identification of its Metabolites[J]. Current Biotechnology, 2025, 15(6): 1040-1049.

图/表 16

图1 ZJ353在不同培养基培养25 d的菌落形态

Fig. 1 Colony morphology of 25 d cultured in different medium of strain ZJ353

图2 ZJ353系统发育树图

Fig. 2 Phylogenetic tree of ZJ353

图3 培养基对发酵粗提物的影响A：培养基对发酵提取物质量的影响；B:不同培养基发酵粗提物对指示菌的抑菌活性

Fig. 3 Effects of culture medium on fermentation crude extract characteristics

图4 加水量对粗提物的影响

Fig. 4 Effect of water addition on crude extract properties

图5 时间对粗提物的影响

Fig. 5 Effect of time on crude extract properties

图6 温度对粗提物的影响

Fig. 6 Effect of temperature on crude extract properties

图7 化合物1的高效液相色谱图

Fig. 7 The high performance liquid chromatography of the compound 1

图8 化合物1对金黄色葡萄球菌的抑菌效果图

Fig. 8 Antibacterial activity of compound 1 against S. aureus

表1 化合物1对金黄色葡萄球菌的最小抑菌浓度测定结果

Table 1 MIC of compound 1 against S. aureus

化合物浓度/(mg·mL^-1)	OD₆₀₀值	化合物浓度/(mg·mL^-1)	OD₆₀₀值
0.156	0.108±0.035	0.009	0.429±0.024
0.078	0.129±0.024	阳性对照	0.698±0.011
0.039	0.145±0.019	阴性对照	0.047±0.017
0.019	0.247±0.031

图9 化合物1正离子扫描质谱图

Fig. 9 Positive ions scanning mass spectrometry of compound 1

图10 化合物1的红外光谱图

Fig. 10 IR spectra of compound 1

图11 化合物1的1H NMR图

Fig. 11 1H nuclear magnetic resonance spectra of compound 1

图12 化合物1的13C NMR图

Fig. 12 13C nuclear magnetic resonance spectra of compound 1

图13 化合物1的化学结构式

Fig. 13 Chemical structure formula of compound 1

表2 化合物1的1H NMR数据

Table 2 1H NMR data for compound 1

位置	δ_H/ppm
aromatic C-H	7.73(s, 1H)
aromatic C-H	7.54(s, 1H)
-O-CH-	4.32(s, 1H)
-CH₂	1.70(s, 1H)
-CH₂	1.43(s, 1H)
-CH₃	0.94(s, 1H)

表3 化合物1的13C NMR数据

Table 3 13C NMR data for compound 1

位置	δ_C/ppm
1	166.0, s
2	132.34, s
3	130.89,s
4	128.84, s
5	65.57, s
6	30.59, s
7	19.20, s
8	13.73, s

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海洋真菌Meira nashicola ZJ353的固态发酵及其代谢产物的分离与鉴定

Solid-state Fermentation of Marine Fungus Meira nashicola ZJ353 and the Isolation and Identification of its Metabolites

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