Variation of Bioactivities and Secondary Metabolomics of Marine Fungus Aspergillus unguis DLEP2008001 Cultured under Different Salinities

doi:10.13560/j.cnki.biotech.bull.1985.2024-0150

Abstract

Abstract:

【Objective】 A marine fungus Aspergillus unguis DLEP2008001 was used for investigating the influence of salinity on the bioactivity and secondary metabolites. 【Method】 The crude extract samples of this strain cultured under different salinities in solid and liquid mediums for 14 and 28 days were analyzed by thin layer chromatography and antioxidative, acetylcholinesterase（AChE）inhibitory, and antimicrobial bioautographies to compare the bioactive secondary metabolites. The secondary metabolite diversity was further investigated by high performance liquid chromatography and “Multi-approach Assisted Feature Based Molecular Network（FBMN）” metabolomic analytical method based on liquid chromatography-tandem mass spectrometry（LC-MS/MS）. 【Result】 The salinity regulated the yield and diversity of antioxidant, AChE inhibitory ingredients and anti-microbial. In seawater-potato-sucrose liquid medium, the strain presented the most fertile feature metabolites when cultured for 28 d under salinity of 35 g/L, while in rice solid medium, it produced the most abundant predominant compounds under salinity of 5 g/L and 35 g/L as the second best condition. The average molecular weights of the predominant metabolites from solid culture generally deceased with the ascending of the salinity and they were mostly small molecules with multiple hydroxyl and amino groups, which are speculated to involve the adjustment of osmotic pressure. 【Conclusion】 Salinity exerts a significant influence on the bioactivity and secondary metabolites of this strain.

Key words: different salinities, marine fungi, molecular network, secondary metabolites, biological activity

KANG Xiao-bo, ZHANG Jing-xi, LU Tian-tian, LIU Ya-yue, ZHOU Long-jian, ZHANG Yi. Variation of Bioactivities and Secondary Metabolomics of Marine Fungus Aspergillus unguis DLEP2008001 Cultured under Different Salinities[J]. Biotechnology Bulletin, 2024, 40(11): 296-311.

Figures/Tables 11

References 44

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培养类型Culture type	培养时间Culture time/d	提取物 Extracts	不同手段展示的主要差异斑点（比移值）及其对应盐度 The main different spots（shown by R_fvalues）displayed by different approaches and the corresponding salinities
培养类型Culture type	培养时间Culture time/d	提取物 Extracts	254 nm紫外光 254 nm ultraviolet	365 nm紫外光 365 nm ultraviolet	抗氧化活性自显影 Antioxidant bioautography	AChE抑制活性自显影 AChE inhibitory bioautography	茴香醛显色 Anisaldehyde coloration	铁氰化钾-三氯化铁显色Potassium ferricyanide-FeCl₃ coloration
液体培养Liquid culture	14	菌丝体提取物 Mycelial extract	—	—	—	R_f（0.6, 0.8）: 10, 15 g/L	R_f（0.2, 1.0）: 2, 5, 10 g/L	—
	14	发酵液提取物 Broth extract	—	R_f（0.6, 1.0）: 5 g/L	R_f（0.4, 0.8）: 0.5, 2, 5 g/L	R_f（0.8, 1.0）: 0, 0.5, 2, 5 g/L	—	—
	28	菌丝体提取物 Mycelial extract	—	—	—	—	R_f（0.0, 1.0）: 5 g/L	—
	28	发酵液提取物 Broth extract	R_f（0.8, 1.0）: 0, 0.5 g/L	—	R_f（0.5）: 0, 0.5 g/L	—	—	—
固体培养Solid culture	14	总提取物 Total extract	R_f（0.4, 1.0）: 5, 10 g/L	—	R_f（0.4, 1.0）: 5, 10 g/L	R_f（0.4, 0.6）: 0, 0.5, 2 g/L R_f（0.6, 0.8）: 5, 10 g/L	R_f（0.0, 1.0）: 5 g/L	R_f（0.4, 1.0）: 5, 10 g/L
固体培养Solid culture	28	总提取物 Total extract	R_f（0.6, 1.0）: 5, 10 g/L	—	R_f（0.2, 0.5）: 5, 10 g/L	R_f（0.8, 1.0）: 5 g/L	R_f（0.0, 1.0）: 5 g/L	R_f（0.6, 0.8）: 5, 10 g/L R_f（0.2, 0.4）: 5 g/L

培养类型Culture type	培养时间Culture time/d	提取物 Extracts	不同手段展示的主要差异斑点（比移值）及其对应盐度 The main different spots（shown by R_fvalues）displayed by different approaches and the corresponding salinities
培养类型Culture type	培养时间Culture time/d	提取物 Extracts	254 nm紫外光 254 nm ultraviolet	365 nm紫外光 365 nm ultraviolet	抗氧化活性自显影 Antioxidant bioautography	AChE抑制活性自显影 AChE inhibitory bioautography	茴香醛显色 Anisaldehyde coloration	铁氰化钾-三氯化铁显色Potassium ferricyanide-FeCl₃ coloration
液体培养Liquid culture	14	菌丝体提取物 Mycelial extract	—	—	—	R_f（0.6, 0.8）: 10, 15 g/L	R_f（0.2, 1.0）: 2, 5, 10 g/L	—
	14	发酵液提取物 Broth extract	—	R_f（0.6, 1.0）: 5 g/L	R_f（0.4, 0.8）: 0.5, 2, 5 g/L	R_f（0.8, 1.0）: 0, 0.5, 2, 5 g/L	—	—
	28	菌丝体提取物 Mycelial extract	—	—	—	—	R_f（0.0, 1.0）: 5 g/L	—
	28	发酵液提取物 Broth extract	R_f（0.8, 1.0）: 0, 0.5 g/L	—	R_f（0.5）: 0, 0.5 g/L	—	—	—
固体培养Solid culture	14	总提取物 Total extract	R_f（0.4, 1.0）: 5, 10 g/L	—	R_f（0.4, 1.0）: 5, 10 g/L	R_f（0.4, 0.6）: 0, 0.5, 2 g/L R_f（0.6, 0.8）: 5, 10 g/L	R_f（0.0, 1.0）: 5 g/L	R_f（0.4, 1.0）: 5, 10 g/L
固体培养Solid culture	28	总提取物 Total extract	R_f（0.6, 1.0）: 5, 10 g/L	—	R_f（0.2, 0.5）: 5, 10 g/L	R_f（0.8, 1.0）: 5 g/L	R_f（0.0, 1.0）: 5 g/L	R_f（0.6, 0.8）: 5, 10 g/L R_f（0.2, 0.4）: 5 g/L

培养方式及时间 Type and time of culture	盐度Salinity/（g·L^-1）	对不同指示菌的抗菌活性强度 Antimicrobial intensity to different indicator strains
培养方式及时间 Type and time of culture	盐度Salinity/（g·L^-1）	BS	SA	EC	PA	CA
液体培养14 d Liquid culture for 14 d	0	-	-	-	-	-
	0.5	-	-	-	-	-
	2	++	-	-	+	+
	5	+++	-	-	++	-
	10	+++	-	-	+	-
	15	-	-	-	-	-
	20	-	-	-	-	-
	35	-	-	-	-	-
液体培养28 d Liquid culture for 28 d	0	-	-	-	-	-
	0.5	-	-	-	-	-
	2	+++	-	-	+	-
	5	+++	-	-	++	-
	10	+++	-	-	+++	-
	15	-	-	-	-	-
	20	-	-	+	-	-
	35	-	-	+	-	-
固体培养14 d Solid culture for 14 d	0	+++	-	-	++	++
	0.5	+++	-	-	++	#
	2	-	-	-	++	#
	5	-	-	-	-	+
	10	-	-	-	-	+
	15	-	-	-	-	+++
	20	-	-	-	-	+++
	35	+++	-	-	++	++
固体培养28 d Solid culture for 28 d	0	+++	-	-	-	+++
	0.5	++	-	+	+	+++
	2	+++	-	+++	+++	++
	5	++	-	++	++	#
	10	+++	-	-	-	#
	15	-	-	-	-	#
	20	-	-	-	-	#
	35	-	-	-	-	+++

培养方式及时间 Type and time of culture	盐度Salinity/（g·L^-1）	对不同指示菌的抗菌活性强度 Antimicrobial intensity to different indicator strains
培养方式及时间 Type and time of culture	盐度Salinity/（g·L^-1）	BS	SA	EC	PA	CA
液体培养14 d Liquid culture for 14 d	0	-	-	-	-	-
	0.5	-	-	-	-	-
	2	++	-	-	+	+
	5	+++	-	-	++	-
	10	+++	-	-	+	-
	15	-	-	-	-	-
	20	-	-	-	-	-
	35	-	-	-	-	-
液体培养28 d Liquid culture for 28 d	0	-	-	-	-	-
	0.5	-	-	-	-	-
	2	+++	-	-	+	-
	5	+++	-	-	++	-
	10	+++	-	-	+++	-
	15	-	-	-	-	-
	20	-	-	+	-	-
	35	-	-	+	-	-
固体培养14 d Solid culture for 14 d	0	+++	-	-	++	++
	0.5	+++	-	-	++	#
	2	-	-	-	++	#
	5	-	-	-	-	+
	10	-	-	-	-	+
	15	-	-	-	-	+++
	20	-	-	-	-	+++
	35	+++	-	-	++	++
固体培养28 d Solid culture for 28 d	0	+++	-	-	-	+++
	0.5	++	-	+	+	+++
	2	+++	-	+++	+++	++
	5	++	-	++	++	#
	10	+++	-	-	-	#
	15	-	-	-	-	#
	20	-	-	-	-	#
	35	-	-	-	-	+++

培养类型Culture type	盐度 Salinity /（g·L^-1）	代谢物编号 No.of metabolite	离子质荷比Mass-to-charge ratio of the precursor ion（m/z）	缀合离子形式Form of adduct ions	分子式 Molecular formula	化合物名称 Compound name	已有报道真菌来源 Previously reported fungal origin	真菌来源文献 Literature for fungal origin
液体培养基 Liquid culture	0.5	化合物1 Compound 1	417.1435	[M+Na]⁺	C₂₁H₂₇ClO₅	Isochromophilones IV	Penicillium multicolor	[21]
液体培养基 Liquid culture	2	化合物2 Compound 2	477.3148	[M+H-H₂O]⁺	C₂₈H₄₆O₇	20-Hydroxyecdysone	Tapinella panuoides	[22]
固体培养基 Solid culture	5	化合物3 Compound 3	304.1362	[M+H]⁺	C₁₃H₂₁NO₇	麦角菌素类 Mycosporin	Gnomonia leptostyla	[23]
		化合物4 Compound 4	152.0088	[M+H]⁺	C₇H₅NOS	2-苯并噻唑酮 2- Benzothiazolone	Dipodascus sp.	[24]
		化合物5 Compound 5	233.1478	[M+H-H₂O]⁺	C₁₅H₂₂O₃	倍半萜类 Sesquiterpenoid	Drechslera gigantea	[25]
		化合物6 Compound 6	169.0424	[M+H]⁺	C₈H₈O₄	苔色酸 Orsellinic acid	Aspergillus sp.	[26-27]
		化合物7 Compound 7	237.1435	[M+H]⁺	C₁₄H₂₂O₄	Gliocladic acid	Trichoderma virens	[28]
	10	化合物8 Compound 8	277.0707	[M+H]⁺	C₁₄H₁₂O₆	Talaroflavone	Talaromyces flavus et al	[29-30]
	35	化合物9 Compound 9	214.0542	[M+H]⁺	C₆H₁₃N₃O₃	瓜氨酸 Citrulline	Schizosaccharomyces pombe	[31]
		化合物10 Compound 10	327.1181	[M+H]⁺	C₁₉H₁₈O₅	Unguinol	Aspergillus unguis	[12,32]
		化合物11 Compound 11	380.1190	[M+H]⁺	C₁₃H₂₁N₃O₈S	S-D-乳酰谷胱甘肽 S-Lactoylglutathione	Saccharomyces cerevisiae	[33]