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

doi:10.19586/j.2095-2341.2025.0072

Current Biotechnology ›› 2025, Vol. 15 ›› Issue (6): 1040-1049.DOI: 10.19586/j.2095-2341.2025.0072

• Articles • Previous Articles Next Articles

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

Abstract

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

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

CLC Number:

Q935

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.

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.

Figures/Tables 16

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

Fig. 2 Phylogenetic tree of ZJ353

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

Fig. 4 Effect of water addition on crude extract properties

Fig. 5 Effect of time on crude extract properties

Fig. 6 Effect of temperature on crude extract properties

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

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

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

Fig. 9 Positive ions scanning mass spectrometry of compound 1

Fig. 10 IR spectra of compound 1

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

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

Fig. 13 Chemical structure formula of compound 1

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)

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

References 28

[1]	VEZZI A, CAMPANARO S, D'ANGELO M, et al.. Life at depth: Photobacterium profundum genome sequence and expression analysis[J]. Science, 2005, 307(5714): 1459-1461.
[2]	王成尧,李雪岩,杨怡侠.780株临床分离多重耐药菌分布及耐药性分析[J].内蒙古医学杂志,2024,56(4):390-396.
	WANG C Y, LI X Y, YANG Y X. Distribution and antibiotic resistance analysis of 780 clinical isolates of multidrug-resistant bacteria[J]. Inn. Mong. Med. J., 2024, 56(4): 390-396.
[3]	PARKES R J, CRAGG B A, WELLSBURY P. Recent studies on bacterial populations and processes in subseafloor sediments: a review[J]. Hydrogeol. J., 2000, 8(1): 11-28.
[4]	赵昌会,叶德赞,魏文铃.深海微生物的研究进展[J].微生物学通报,2006,33(3):142-146.
	ZHAO C H, YE D Z, WEI W L. Research on deep-sea microbiology[J]. Microbiol. China, 2006, 33(3): 142-146.
[5]	陈皓文,孙丕喜,高爱国.微生物学向深海进军[J].海洋地质动态,2009,25(2):14-20.
	CHEN H W, SUN P X, GAO A G. Microbiology marches into the deep sea[J]. Mar. Geol. Lett., 2009, 25(2): 14-20.
[6]	BULL A T, STACH J E M. Marine actinobacteria: new opportunities for natural product search and discovery[J]. Trends Microbiol., 2007, 15(11): 491-499.
[7]	ISHII A, NAKASONE K, SATO T, et al.. Isolation and characterization of the dew cluster from the piezophilic deep-sea bacterium Shewanella violacea [J]. J. Biochem., 2002, 132(2): 183-188.
[8]	KATO C, NOGI Y. Correlation between phylogenetic structure and function: examples from deep-sea Shewanella [J]. FEMS Microbiol. Ecol., 2001, 35(3): 223-230.
[9]	杨加庚,梅益勤,裴月湖.海洋真菌次级代谢产物化学成分及生物活性的研究进展[J].沈阳药科大学学报,2013,30(1):72-82.
	YANG J G, MEI Y Q, PEI Y H. Research progress on the second metabolites produced by marine fungi and their bioactivities[J]. J. Shenyang Pharm. Univ., 2013, 30(1): 72-82.
[10]	朱伟明,王俊锋.海洋真菌生物活性物质研究之管见[J].菌物学报,2011,30(2):218-228.
	ZHU W M, WANG J F. A review on studies of secondary metabolites from marine fungi[J]. Mycosystema, 2011, 30(2): 218-228.
[11]	SCHUEFFLER A, ANKE T. Fungal natural products in research and development[J]. Nat. Prod. Rep., 2014, 31(10): 1425-1448.
[12]	CHEN G, WANG H F, PEI Y H. Secondary metabolites from marine-derived microorganisms[J]. J. Asian Nat. Prod. Res., 2014, 16(1): 105-122.
[13]	耐甲氧西林金黄色葡萄球菌感染防治专家委员会.耐甲氧西林金黄色葡萄球菌感染防治专家共识2011年更新版[J].中华实验和临床感染病杂志(电子版),2011,5(3):372-384.
	Expert consensus on prevention and treatment of methicillin-resistant Staphylococcus aureus infection: 2011 update[J]. Chin. J. Exp. Clin. Infect. Dis. Electron. Ed., 2011, 5(3): 66-72.
[14]	CAMPANARO S, VEZZI A, VITULO N, et al.. Laterally transferred elements and high pressure adaptation in Photobacterium profundum strains[J/OL]. BMC Genom., 2005, 6(1): 122[2025-11-06]. .
[15]	OTTO M. Staphylococcus aureus toxins[J]. Curr. Opin. Microbiol., 2014, 17: 32-37.
[16]	MAMALIS N. Ocular methicillin-resistant Staphylococcus aureus [J]. J. Cataract Refract. Surg., 2014, 40(11): 1757-1758.
[17]	TARMAN K, LINDEQUIST U, WENDE K, et al.. Isolation of a new natural product and cytotoxic and antimicrobial activities of extracts from fungi of Indonesian marine habitats[J]. Mar. Drugs, 2011, 9(3): 294-306.
[18]	魏景超遗.真菌鉴定手册[M].上海:上海科学技术出版社,1979.
[19]	庄文颖. 中国真菌志[M]. 北京:科学出版社,2004.
[20]	邵律成,盛春泉,张万年.新结构类型抗真菌先导化合物的研究进展[J].药学学报,2007,42(11):1129-1136.
	SHAO L, SHENG C Q, ZHANG W N. Recent advances in the study of antifungal lead compounds with new chemical scaffolds[J]. Acta Pharm. Sin., 2007, 42(11): 1129-1136.
[21]	刘晴晴,叶晓洁,齐然,等.邻苯二甲酸二丁酯的GC检测及其合成动力学研究[J].皮革与化工,2020,37(2):19-22.
	LIU Q Q, YE X J, QI R, et al.. Study on the synthesis of dibutyl phthalate by gas chromatography[J]. Leather Chem., 2020, 37(2): 19-22.
[22]	马桂珍,付泓润,吴少杰,等.海洋多粘类芽孢杆菌L1-9抗菌活性产物的分离与结构鉴定[J].植物病理学报,2014,44(5):486-496.
	MA G Z, FU H R, WU S J, et al.. Isolation and structure elucidation of antifungal metabolites from marine Paenibacillus polymyxa strain L1-9[J]. Acta Phytopathol. Sin., 2014, 44(5): 486-496.
[23]	刘伟,徐涛,蔡敬民,等.海洋小链霉菌DY2741抗菌物质的溶解性质及分离纯化[J].食品科学,2010,31(15):177-180.
	LIU W, XU T, CAI J M, et al.. Isolation, purification and solubility of antibacterial substances from fermentation supernatant of marine Streptomyces parvus DY2741[J]. Food Sci., 2010, 31(15): 177-180.
[24]	BI Y, LIU G, YU Q, et al.. Anti-Vibrio dibutyl phthalate from marine-derived Streptomyces sp. S073[J]. Res. Vet. Sci., 2021, 140: 198-202.
[25]	李元鹏,牟晓玲,马家易,等.邻苯二甲酸二丁酯对镰刀菌化感抑制机制研究[J].甘肃农业大学学报,2023,58(5):113-121.
	LI Y P, MU X L, MA J Y, et al.. Allelopathy inhibition and its mechanism by dibutylphthalate on Fusarium [J]. J. Gansu Agric. Univ., 2023, 58(5): 113-121.
[26]	孙正昊.邻苯二甲酸二丁酯对黑土微生物群落及大豆生长发育的影响[D].哈尔滨:中国科学院大学,2024.
[27]	王沛芳,张艺璇,董跃,等.邻苯二甲酸二丁酯对电活性生物膜性能影响及机制[J].河海大学学报(自然科学版),2021,49(2):113-120.
	WANG P F, ZHANG Y X, DONG Y, et al.. Effect and mechanism of dibutyl phthalate on performance of electroactive biofilm[J]. J. Hohai Univ. Nat. Sci., 2021, 49(2): 113-120.
[28]	焦裕怀.邻苯二甲酸二异丁酯暴露对蚯蚓的氧化损伤及其作用机理研究[D].泰安:山东农业大学,2021.

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

RichHTML

PDF

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 16

References 28

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	Tieliang ZHAO, Zhongying JIANG, Mingyun SHI. Research Progress on the Expansion and Rupture of Giant Unilamellar Vesicles Under Osmotic Stress [J]. Current Biotechnology, 2025, 15(5): 791-797.
[2]	Lulu ZHAO, Tian HONG, Yiran HAO, Erning CHEN, Jingwen LI, Meihong DU. Research Progress of Immunomagnetic Separation Technology in Detection of Circulating Tumor Cell [J]. Current Biotechnology, 2025, 15(4): 606-614.
[3]	Haixia PENG, Shijuan WANG, Zhuanxia XIN, Li MEI, Meng MA. A Low-cost, Efficient and Easy-to-operate Method for Transgenic Plant Identification [J]. Current Biotechnology, 2024, 14(6): 1016-1023.
[4]	Haoran DONG, Ning JIANG, Huan LU, Yang FU, Qiaozhen LI, Hailong YU. Research Progress on Structure and Function of Lentinan [J]. Current Biotechnology, 2024, 14(6): 911-919.
[5]	Shaoting PAN, Boxuan WANG, Jiaxin CHEN, Jiajun CAI, Yanshen LIN, Lingzhi TANG, Xuan HONG. Research Progress on Polyketides from Marine Fungi [J]. Current Biotechnology, 2024, 14(6): 993-1003.
[6]	Guizhen GAO, Yungu ZHAI, Lubin ZHANG, Jinmei CHANG, Haihua LUO, Xiaoming WU. Advances on Germplasm Identification and Variety Breeding to Saline-alkali Stress of Rapeseed [J]. Current Biotechnology, 2022, 12(5): 647-654.
[7]	Wenjing ZHANG, Ye TONG, Xiwen YANG, Yanjin CAO, Jidong WEI. Screening and Optimization of Highyield Neutral Proteaseproducing Strains and Pilot Scaleup [J]. Current Biotechnology, 2022, 12(1): 112-119.
[8]	Rong XIAO, Yunxiao WEI, Yuan WANG, Zhigang MENG, Chengzhen LIANG, Quanjia CHEN, Rui ZHANG. Exploring of Authenticity Identification Method of Transgenic Cotton Plant by Shoot Tip [J]. Current Biotechnology, 2022, 12(1): 83-89.
[9]	Nana ZHOU, Xiaoyan WANG, Yuan ZHANG, Jing WANG, Guomiao ZHAO, Chao WEI, Kai YANG, Tai AN. Progress on the Production Technology of Recombinant Therapeutic Proteins [J]. Current Biotechnology, 2021, 11(6): 724-731.
[10]	Wenling ZHAI, Caiyun LIU, Ying LIU, Bisheng FU, Jin CAI, Wei GUO, Qiaofeng ZHANG, Jizhong WU. Phenotypic and Molecular Identification of New Wheat Germplasm Resistant to Fusarium Head Blight [J]. Current Biotechnology, 2021, 11(5): 581-589.
[11]	GAO Jiaqi1, CHEN Shuo1, WANG Di2, LONG Yan2, LI Liang2, ZHANG Xiao1. Progress on Development and Detection of Internal Reference Genes in Six Crops [J]. Curr. Biotech., 2020, 10(6): 613-622.
[12]	CHEN Shuo1, GAO Jiaqi1, WANG Di2, LONG Yan2, LI Liang2, ZHANG Xiao1. DNA Tetrahedral Nanostructure and its Application Progress in Biotechnology [J]. Curr. Biotech., 2020, 10(6): 661-667.
[13]	ZHONG Caixia¹, TIAN Jiaxue², WANG Xiaoqi², WEN Tong^2*. Isolation, Identification of an Acetic Acid Producing Strain from Tomato Surface and the Optimization of Acid Producing Conditions [J]. Curr. Biotech., 2020, 10(5): 564-569.
[14]	LI Xuyang, HUANG Liling, GUO Qiannan, GAO Ruyu, ZHANG Wei, CHEN Ming, LU Wei, ZHOU Zhengfu*. Isolating, Screening and Identification Involved in 2,4-D Degradation [J]. Curr. Biotech., 2019, 9(4): 384-395.
[15]	CHEN Zeli1, YANG Linyi2, CHEN Lu2, SUN Yan2, WEI Zhaoxia2, LI Yongzhong3, ZHAO Mingfu2, WEN Guosong1. Detection and Identification of BCMV Infecting Polygonatum kingianum [J]. Curr. Biotech., 2019, 9(2): 122-128.