文献类型：期刊文献

英文题名：The screening for marine fungal strains with high potential in alkaloids production by in situ colony assay and LC-MS/MS based secondary metabolic profiling

作者：Lu, Tiantian[1];Liu, Yayue[1,2,3];Zhou, Longjian[1,2,3];Liao, Qingnan[1];Nie, Yingying[1];Wang, Xingyuan[1];Lei, Xiaoling[1,2,3];Hong, Pengzhi[1,2,3];Feng, Yan[1];Hu, Xueqiong[1,2,3];Zhang, Yi[1,2,3]

机构：[1]Guangdong Ocean Univ, Shenzhen Inst, Coll Food Sci & Technol, Guangdong Prov Engn Technol,Research Ctr Seafood, Zhanjiang, Peoples R China;[2]Southern Marine Sci & Engn Guangdong Lab Zhanjiang, Zhanjiang, Peoples R China;[3]Dalian Polytech Univ, Prov Minist Collaborat Innovat Ctr Key Technol Mar, Dalian, Peoples R China

年份：2023

卷号：14

外文期刊名：FRONTIERS IN MICROBIOLOGY

收录：SCI-EXPANDED(收录号：WOS:000990007700001)、、WOS

基金：This study was supported by the Guangdong Provincial Natural Science Foundation (Grant Number: 2022A1515010783), the Guangdong Provincial Special Project in Science and Technology (Grant Number: 2021A05240), the National Natural Science Foundation (Grant Number: 21807015), the Special project in key fields of Guangdong provincial higher education institutions (Biomedicine and health care) (Grant Number: 2021ZDZX2064), the Basic Research Project of Shenzhen Science and Technology Innovation Commission (Grant Numbers: JCYJ20220530162014032 and JCYJ20190813105005619), the Zhanjiang Marine Youth Talent Innovation Project (Grant Number: 2022E05010), the Shenzhen Dapeng New District Scientific and Technological Research and Development Fund (Grant Number: KJYF202001-07), the Science and Technology Development Special Fund Competitive Allocation Project of Zhanjiang City (Grant Number: 2020A01030), the Program for Scientific Research Start-up Funds of Guangdong Ocean University (Grant Numbers: R18008 and 060302042201), the Innovation and Development Project about Marine Economy Demonstration of Zhanjiang City (Grant Number: Zhan-Hai-Chuang XM-202008-01B1), and the Graduate Student Education Innovation Project of Guangdong Ocean University (Grant Number: 202232).

语种：英文

外文关键词：alkaloids; in situ colony screening; LC-MS/MS; Penicillium mallochii; azaphilone; anti-neuroinflammatory

外文摘要：Background: Alkaloids are the second primary class of secondary metabolites (SMs) from marine organisms, most of which have antioxidant, antitumor, antibacterial, anti-inflammatory, and other activities. However, the SMs obtained by traditional isolation strategies have drawbacks such as highly reduplication and weak bioactivity. Therefore, it is significantly important to establish an efficient strategy for screening strains and mining novel compounds. Methods: In this study, we utilized in situ colony assay combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS) to identify the strain with high potential in alkaloids production. The strain was identified by genetic marker genes and morphological analysis. The secondary metabolites from the strain were isolated by the combine use of vacuum liquid chromatography (VLC), ODS column chromatography, and Sephadex LH-20. Their structures were elucidated by 1D/2D NMR, HR-ESI-MS, and other spectroscopic technologies. Finally, these compounds bioactivity were assay, including anti-inflammatory and anti-beta aggregation. Results: Eighteen marine fungi were preliminarily screened for alkaloids production by in situ colony assay using Dragendorff reagent as dye, and nine of them turned orange, which indicated abundant alkaloids. By thin-layer chromatography (TLC), LC-MS/MS, and multiple approaches assisted FeatureBased Molecular Networking (FBMN) analysis of fermentation extracts, a strain ACD-5 (Penicillium mallochii with GenBank accession number OM368350) from sea cucumber gut was selected for its diverse alkaloids profiles especially azaphilones. In bioassays, the crude extracts of ACD-5 in Czapek-dox broth and brown rice medium showed moderate antioxidant, acetylcholinesterase inhibitory, anti-neuroinflammatory, and anti-b aggregation activities. Three chlorinated azaphilone alkaloids, compounds 1-3 (sclerotioramine, isochromophilone VI, and isochromophilone IX, respectively), were isolated from the fermentation products of ACD-5 in brown rice medium guided by bioactivities and mass spectrometry analysis. Compound 1 had shown remarkable anti-neuroinflammatory activity in liposaccharide induced BV-2 cells. Conclusion: In summary, in situ colony screening together with LC-MS/MS, multi-approach assisted FBMN can act as an efficient screening method for strains with potential in alkaloids production.