Bioactive chemical constituents from the marine-derived fungus Cladosporium sp.DLT-5

A new isochromanone,cladosporinisochromanone(1),accompanied by 15 known compounds(2–16)were obtained from secondary metabolites produced by marine-derived fungus Cladosporium sp.DLT-5.NMR and HRESIMS spectra elucidation determined the planar structure of 1.Subsequent electronic circular dichroism(ECD)experiment assigned the absolute configuration of 1.Compounds 1,2,4–6,and 10 displayed different degrees of neuroprotective activities on human neuroblastoma cells SH-SY5Y.Five compounds(1,3–5,and 13)emerged resistance to protein tyrosine phosphatase 1B(PTP1B),further kinetic analysis and molecular docking study indicated that the most potent compound 13(IC50value of 10.74±0.61μmol/L)was found as a noncompetitive inhibitor for PTP1B.Surface plasmon resonance(SPR)and molecular docking studies also demonstrated the interaction between compound 12 and Niemann-Pick C1 Like 1(NPC1L1),which has been identified as significant therapeutic target for hypercholesteremia.In addition,compounds 3,6,and 14 showed attractive inhibitory activity against the phytopathogenic fungi:Colletotrichum capsici.Therefore,library of Cladosporium metabolites is enriched and new active uses of known compounds are explored.

The Antitumor Components from Marine-derived Bacterium Streptoverticillium luteoverticillatum 11014 Ⅱ

Eight known compounds were isolated from a marine-derived bacterium Streptoverticillium luteoverticillatum 11014 using bioassay-guided fractionations. Their structures were identified by spectral analysis as bis （4-hydroxybenzyl） ether （1）, p-hydroxyphenylethyl alcohol （2）, N-（4-hydroxyphenethyl） acetamide （3）, indole-3 carboxylic acid methyl ester （4）, dibenzo[b,e] [1,4]dioxine （5）, thymine （6）, cytosine deoxyribonucleoside （7） and 2, 3-butanediol （8）. These compounds were evaluated for their cytotoxic activities against K562 cell line with the SRB method for the first time. Compounds 2 and 4 showed cytotoxcities with IC50 values of 101.1 and 165.3 μolL^-1, respectively. All compounds were isolated from S. luteoverticillatum 11014 for the first time.

The Cytotoxic Constituents from Marine-derived Streptomyces 3320^(#)

The present work studies the chemical constituents from marine-derived streptomyces 3320^# and their antitumor activities. The n-BuOH extract of the ferment broth of 3320^# was chromatographed on silica gel, Sephadex LH-20, ODS columns and HPLC to separate the compounds with antitoumor activities. Their structures were identified using IR, UV, NMR, MS spectroscopic techniques and compared with published data. The antitumor activities of the isolates were assayed using SRB method and flow cytometry assay, accompanied with the morphological observation of the cells under light micro- scope against mammalian tsFT210 cells. Ten compounds, cyclo-（Ala-Leu） 1, cyclo-（Ala-Ile） 2, cyclo-（Ala-Val） 3, cyclo- （Phe-Pro） 4, cyclo-（Phe-Gly）5, cyclo-（Leu-Pro）6, 1-methyl-1, 2, 3, 4-tetrahydro-β-carboline-3-carboxylic acid 7, N-（4- hydroxyphenethyl） acetamide 8, 4-methyoxy-l-（2-hydroxy） ethylbenzene 9 and uridine 10, were isolated from the ferment broth of streptomyces 3320^# . Among them, compounds 6, 7, 8 and 10 showed potent cytotoxicity against the tsFT210 cell with the IC50 values of 3.6, 7.2, 5.2 and 1.6 mmol L-1, respectively. Compounds 8, 10 also exhibited apoptosis inducing activity under 2.0 mmol L-1. Compounds 6, 7, 8 and 10 are the principle bioactive constituents responsible for the antitumor activities of marine streptomyces 3320^#. Compound 7 was isolated from this species for the first time.

High Ethanol Production by Marine-Derived Yeasts-<i>Saccharomyces cerevisiae</i>under Stress Pressures

For practical applications of bioethanol, the uses of both highly concentrated biomass materials and their effective fermentation by yeasts are indispensable in order to produce ethanol at low costs. However, as the saccharified products of those biomass generally contain abundant sugars, the yeasts are affected by the compounds and are inclined to decrease their physiological activities. In the process of fermentation, ethanol is gradually produced by the yeasts in the culture;the concentrated metabolic product also damages itself, and inhibition of the fermentation frequently occurs. The application of yeasts with high fermentative activities under stress pressures such as sugars and ethanol is thus desired for bioethanol production. In this study, various types of high-fermentative yeasts under stress pressures were isolated mainly from coastal waters in Japan and characterized. All yeast strains with high fermentative activities under 20% v/v ethanol were found to be Saccharomyces cerevisiae. The HK21 strain isolated from Tokyo Bay and identified as S. cerevisiae had the highest fermentation activity under 30% w/v sorbitol and under 20% v/v ethanol, and it produced approx. 70 g/l (9% v/v) ethanol from the 15% w/v glucose solution at 25 oC within 5 days.

Strain and Culture Medium Optimization for Production Enhancement of Prodiginines from Marine-Derived Streptomyces sp. GQQ-10

A mutant (GQQ-M6) of a Sponge-Derived streptomyces sp. GQQ-10 obtained by UV-induced mutation was used for producing prodiginines (PGs). Single factor experiments and orthogonal array design (OAD) methods were employed for medium optimization. In the single factor method, the effects of soluble starch, glucose, soybean flour, yeast extract and sodium acetate on PGs production were investigated individually. In the subsequent OAD experiments, the concentrations of these 5 key nutritional components combined with salinity were further adjusted. The mutant strain GQQ-M6 gave a 2.2-fold higher PGs production than that of the parent strain; OAD experiments offered a PGs yield of 61mg L-1, which was 10 times higher than that of the initial GQQ-10 strain under the original cultivation mode.

放线菌19G-317菌株发酵产物抑菌活性初步研究

【目的】明确放线菌19G-317菌株发酵产物对植物病原菌的抑制活性,为该菌株在农业病害防治中的开发应用及后续研究奠定基础。【方法】采用生长速率法、孢子萌发法、活体组织法和盆栽试验,测定放线菌19G-317菌株发酵产物的抑菌效果。【结果】生长速率法试验表明,放线菌19G-317菌株发酵产物对供试22种植物病原菌菌丝生长均有不同程度的抑制作用,发酵液对其中11种植物病原菌的抑制率大于60%,菌丝提取物对其中8种植物病原菌的抑制率大于90%;发酵液和菌丝提取物对油菜菌核病菌菌丝生长的EC50分别为84.30和18.80 mg/L。孢子萌发试验表明,发酵液和菌丝提取物对番茄灰霉病菌孢子的萌发均有较强的抑制作用,在0.5 mg/mL质量浓度下,其抑制率分别为94.26%和96.20%;组织法试验表明,在10 mg/mL质量浓度下,发酵液和菌丝提取物对番茄灰霉病的药效分别为72.17%和76.95%;盆栽试验表明,19G-317菌株发酵液和菌丝提取物对辣椒疫霉病均有一定的保护和治疗作用。【结论】放线菌19G-317菌株抑菌谱较广,对植物病菌的防治有很大的研究开发潜力。

抗肿瘤活性海洋放线菌的筛选及菌株HGF26的初步鉴定

对采自连云港海域的海泥样品进行放线菌选择性分离,用其发酵液进行抗肿瘤活性筛选,并对活性较好的菌株HGF26进行了初步鉴定。从海泥样品中共分离得到放线菌78株,以人肝癌细胞HepG2为靶标,活性筛选得到细胞毒活性达60%以上的阳性菌株3株,以其他5株肿瘤细胞为靶标的复筛表明,菌株HGF、26的发酵液对多种肿瘤细胞具有显著的细胞毒活性,其中对胃癌细胞BGC823的细胞毒活性为79%,活性产物具有较好的酸碱和热稳定性。通过对菌株的培养特征、形态特征、生理生化特征和细胞壁成分分析,将菌株HGF26初步鉴定为微白黄链霉菌的海洋变种。

抗多重耐药金黄色葡萄球菌土壤放线菌筛选

[目的]从土壤中发现拮抗多重耐药金黄色葡萄球菌的放线菌。[方法]将土壤稀释后在高氏一号培养基上划线分离放线菌,并测试抗菌活性。[结果]获得放线菌一株B0602具有很强的抗菌活性。[结论]放线菌B0602可能开发出高活性的抗生素制品。

抑制黄瓜蔓枯菌的放线菌筛选

以黄瓜蔓枯菌为指示菌,采用平板稀释法和对峙法获得拮抗放线菌菌株9株,并以常见的10种蔬菜病原菌为目标菌进行复筛,发现菌株A9、T111拮抗谱较宽,对黄瓜蔓枯菌(Ascochyta citrullina Smith)、黄瓜枯萎菌(Fusarium.oxysporum)、黄瓜菌核((Sclerotinia scleotiorum)等有很好的抑制作用,并具有产几丁质酶的能力。经形态、培养特性等初步鉴定为链霉菌属(Streplomyces)不同类群。A9、T111发酵液在病原菌活体定植抑制测定试验中,对黄瓜蔓枯病菌抑制效果分别达56.7%和65.4%。

新疆南部土壤放线菌区系研究

从新疆南部巴州和克州两地区8个县(市)采集农田、原始森林、草甸、戈壁、荒漠和盐碱土壤51份,使用6种分离培养基,9种分离方法分离,并对分离得到的放线菌进行了鉴定。共得到9个属的放线菌,其中链霉菌为优势菌,并可分为13个类群。同时,较为全面、系统地研究了两地区之间和不同类型土壤中放线菌区系组成和异同。

海洋放线菌抗菌活性菌株的筛选及南-148菌株的鉴定

以桉树青枯病菌和金黄色葡萄球菌为指示菌,对从广东湛江近海海泥中分离得到的135株放线菌的抗菌活性进行测定,并对南-148菌株进行形态观察和生理生化特性鉴定。结果表明,135株海洋放线菌中有15株对桉树青枯病菌有抑制作用,24株对金黄色葡萄球菌有抑制作用,12株对两种病菌均有抑制作用,其中南-148菌株的抑菌效果最好,对桉树青枯病菌的抑菌圈直径为20.0mm,对金黄色葡萄球菌的抑菌圈直径为37.3mm;对南-148菌株进行了鉴定,初步确定南-148菌株为弗氏链霉菌(Streptomyces fradiae)。

Lj20和St24发酵液粗提物对番茄体内防御性酶活性的影响

为明确植物内生放线菌对植物体内防御性酶活性的影响,以植物内生放线菌Lj20和St24发酵液粗提物处理番茄植株,测定其根、茎、叶中过氧化氢酶(CAT)、多酚氧化酶(PPO)、过氧化物酶(POD)和超氧化物歧化酶(SOD)的活性变化。测定结果表明,用Lj20和St24粗提物处理番茄植株后,除SOD处理和对照没有明显差异外,CAT、PPO和POD酶的活性都有所增强,但酶活性随时间变化规律各不相同。其中Lj20粗提物处理的番茄茎和叶中PPO的活性增加最明显,在测定时间内呈直线上升趋势。这表明Lj20和St24的代谢物不仅对病原菌具有直接抑制作用,而且可通过提高植物体内防御性酶活性来提高植物的抗病性。

Establishment of an efficient transformation protocol and its application in marine-derived Bacillus strain

Marine-derived Bacillus strains have been proved to be a very promising source for natural product leads.However,transformation of environmental strains is much more difficult than that of domesticated strains.Here,we report the development of an efficient and robust electroporation-based transformation system for marine-derived Bacillus marinus B-9987,which is a macrolactin antibiotics producer and a very promising biological control agent against fungal plant diseases.The transformation efficiency was greatly enhanced 103-fold by using unmethylated plasmid to bypass modification-restriction barrier,and using glycine betaine to protect cells from electrical damages during electroporation.Addition of HEPES and 2 mmol L?1MgCl2 further improved the efficiency by additional 2-fold,with a maximum value of 7.1×104 cfu/μg pHT3101.To demonstrate the feasibility and efficiency of the protocol,a green fluorescent protein reporter system was constructed;furthermore,phosphopantetheinyl transferase gene sfp,which is essential to the biosynthesis of polyketides and nonribosomal peptides,was overexpressed in B-9987,leading to increased production of macrolactin A by about 1.6-fold.In addition,this protocol is also applicable to marine-derived Bacillus licheniforms EI-34-6,indicating it could be a reference for other undomesticated Bacillus strains.To our knowledge,this is the first report regarding the transformation of marine-derived Bacillus strain.

Isolation and characterization of cyclic lipopeptides from the marine-derived Bacillus licheniformis

Chromatographic separation of the marine-derived bacterium Bacillus licheniformis resulted in the isolation of two new cyclic lipopeptides named ai-Cl6 surfactin （1） and ai-Cl4 surfactin （2）, together with iso-Cm5 surfactin and iso-Cl6 surfactin. The structures of the new cyclic lipopeptides were determined through extensive spectroscopic analysis. The sequences of the amino acids in cyclic nucleus were established by the ESI-MS/MS fragmentation, which provided an efficient method to detect lipopeptides from bacterium extracts without senaration

Penicacids E-G, three new mycophenolic acid derivatives from the marine-derived fungus Penicillium parvum HDN17-478

Three new mycophenolic acid derivatives, penicacids E-G(1-3), together with three known analogues, mycophenolic acid(4), 4′-hydroxy-mycophenolic acid(5) and mycophenolic methyl ester(6), were isolated from a marine-derived fungus Penicillium parvum HDN17-478 from a South China Sea marine sediment sample. The structures of compounds 1-3 were elucidated by HRMS, NMR, and Mosher’s method. Among them, compounds 1 and 2 were the first examples of mycophenolic acid analogs with a double bond at C-3′/C-4′ position. The cytotoxicity of 1-6 was evaluated against the HCT-116, BEL-7402, MGC-803, SH-SY5 Y, HO-8910 and HL-60 cell lines, and compounds 4 and 6 showed potent cytotoxicity with IC50 values ranging from 1.69 to 12.98 μmol·L–1.

Burial fluxes and source apportionment of carbon in culture areas of Sanggou Bay over the past 200 years

In this study, we assessed the burial fluxes and source appointment of different forms of carbon in core sediments collected from culture areas in the Sanggou Bay, and preliminarily analyzed the reasons for the greater proportion of inorganic carbon burial fluxes （BFTIc）. The average content of total carbon （TC） in the Sanggou Bay was 2.14%. Total organic carbon （TOC） accounted for a small proportion in TC, more than 65% of which derived from terrigenous organic carbon （Ct）, and while the proportion of marine-derived organic carbon （Ca） increased significantly since the beginning of large-scale aquaculture. Total inorganic carbon （TIC） accounted for 60%-75% of TC, an average of which was 60%, with a maximum up to 90% during flourishing periods （1880-1948） of small natural shellfish derived from seashells inorganic carbon （SheU-IC）. The TC burial fluxes ranged from 31 g/（m2.a） to 895 g/（m2.a） with an average of 227 g/（m2.a）, which was dominated by TIC （about 70%）. Shell-IC was the main source of TIC and even TC. As the main food of natural shellfish, biogenic silica （BSi） negatively correlated with BFTIc through affecting shellfish breeding. BFTIc of Sta. S1, influenced greatly by the Yellow Sea Coastal Current, had a certain response to Pacific Decadal Oscillation （PDO） in some specific periods.

Chemical diversity and biological function of indolediketopiperazines from marine-derived fungi

Natural products from marine-derived fungi have attracted considerable attention in the recent two decades.Indolediketopiperazines are one of the most important classes of marine natural products,mainly discovered from the fungal genera Penicillium,Aspergillus and Eurotium.These compounds span a wide range of chemical structures and bioactivities.This review summarizes 155 indolediketopiperazines that were discovered from marine-derived fungi from 2000 to early 2019 and primarily focuses on their chemical diversity and biological function.

Safe antifungal lipopeptides derived from Bacillus marinus B-9987 against grey mold caused by Botrytis cinerea

Agricultural application studies, including field experiments and acute toxicity tests, were conducted for lipopeptides secreted by marine-derived Bacillus marinus B-9987. Benefiting from commercially available scaled-up lipopeptide purification, the sample of impurities（isolated from target lipopeptides）, raw extracted sample（purity： 9.08%）, partially purified sample（purity： 20.86%）, and highly purified sample（purity： 87.51%） were prepared from B. marinus B-9987 fermentation broth, and used in lab-scale antagonism tests, field experiments, swarming motility tests, and acute toxicity tests. Operations and conditions in field experiments were consistent with the Pesticide-Guidelines for the Field Efficacy Trials（GB/T 17980.28-2000）, and acute toxicity tests were executed according to Toxicological Test Methods of Pesticides for Registration（GB 15670-1995）. In agar diffusion tests in vitro and pot tests in vivo, all lipopeptide samples with different purities significantly inhibited Botrytis cinerea; meanwhile the sample of impurities isolated from target lipopeptides were not effective against B. cinerea. Results of lab-scale tests showed that the target lipopeptides were effective substances against B. cinerea. Thus, partially purified and raw extracted samples were used in field experiments instead of the highly purified sample for cost saving. In the field experiments against rose grey mold, biological control efficacy of 500 mg L–1 lipopeptides reached 67.53%, slightly lower than 74.05% reached by the agrochemical pyrimethanil. However, pyrimethanil severely suppressed B. marinus B-9987, whereas the lipopeptides promoted swarming motility and biocontrol efficacy of Bacillus biomass. Lipopeptides at 87.51% purity were tested for systemic acute toxicity and confirmed as low-toxicity substances. In conclusion, low-toxicity lipopeptides were potential alternatives to agrochemicals, and they also performed good promotion when combined with homologous biological control microorganism. There were 2 breakthroughs in this research：（1） marine-derived bacterial lipopeptides inhibited grey mold caused by B. cinerea in field experiments; and（2） purified bacterial lipopeptides（sample purity： 〉87.51%） were determined to be low-toxicity substances by systemic acute toxicity tests, satisfying the strict requirement of pesticide registration in China（required purity： 〉85%）. This study provides support for using extracellularBacillus-derived lipopeptides commercially similar to Bacillus-based biological control agents.

Marine-derived fungi as a source of bioactive indole alkaloids with diversifed structures

Marine-derived fungi are well known as rich sources of bioactive natural products.Growing evidences indicated that indole alkaloids,isolated from a variety of marine-derived fungi,have attracted considerable attention for their diverse,challenging structural complexity and promising bioactivities,and therefore,indole alkaloids have potential to be pharmaceutical lead compounds.Systemic compilation of the relevant literature.In this review,we demonstrated a comprehensive overview of 431 new indole alkaloids from 21 genera of marine-derived fungi with an emphasis on their structures and bioactivities,covering literatures published during 1982–2019.

Polysubstituted Phenyl Glucosides Produced by the Fungus Metarrhizium anisopliae

MetarhizosidesA-G(1-7),seven new polysubstituted phenyl glucosides,were isolated from the extracts of solid rice medium of a marine-derived fungus Metarrhizium anisopliae.Compounds 1-7 all contain a polysubstituted phenyl group and the sugar unit is identified as 4'-O-methyl-β-D-glucopyranose.Their structures were elucidated by NMR spectroscopy and chemical method.These compounds were evaluated for anti-inflammatory activity by using LPS-stimulated murine macrophage RAW 264.7 cells and the cytotoxicities against four human cancer cell lines.