Investigation of Deep-Sea Ecosystems Using Marker Fatty Acids: Sources of Essential Polyunsaturated Fatty Acids in Abyssal Megafauna

Abyssal seafloor ecosystems cover more than 50% of the Earth’s surface. Being formed by mainly heterotrophic organisms, they depend on the flux of particulate organic matter (POM) photosynthetically produced in the surface layer of the ocean. As dead phytoplankton sinks from the euphotic to the abyssal zone, the trophic value of POM and the concentration of essential polyunsaturated fatty acids (PUFA) decrease. This results in pronounced food periodicity and limitations for bottom dwellers. Deep-sea invertebrate seston eaters and surface deposit feeders consume the sinking POM. Other invertebrates utilize different food items that have undergone a trophic upgrade, with PUFA synthesized from saturated and monounsaturated FA. Foraminifera and nematodes can synthesize arachidonic acid (AA), eicosapentaenoic acid (EPA), while some barophylic bacteria produce EPA and/or docosahexaenoic acid. FA analysis of deep-sea invertebrates has shown high levels of PUFA including, in particular, arachidonic acid, bacterial FA, and a vast number of new and uncommon fatty acids such as 21:4(n-7), 22:4(n-8), 23:4(n-9), and 22:5(n-5) characteristic of foraminifera. We suppose that bacteria growing on detritus having a low trophic value provide the first trophic upgrading of organic matter for foraminifera and nematodes. In turn, these metazoans perform the second-stage upgrading for megafauna invertebrates. Deep-sea megafauna, including major members of Echinodermata, Mollusca, and Polychaeta display FA markers characteristic of bacteria, foraminifera, and nematodes and reveal new markers in the food chain.     

Characterization of Bioactivities and Biosynthesis of Angucycline/Angucyclinone Derivatives Derived from Gephyromycinifex aptenodytis gen. nov., sp. nov.

Strain NJES-13^T is the type strain and currently the only species of the newly established actinobacteria genera Aptenodytes in the family Dermatophilaceae isolated from the gut microbiota of the Antarctic emperor penguin. This strain demonstrated excellent bioflocculation activity with bacteria-derived exopolysaccharides (EPSs). Moreover, it produced bioactive angucycline/angucyclinone derivatives (ADs) and contained one type III polyketide synthase (T3PKS), thus demonstrating great potential to produce novel bioactive compounds. However, the low productivity of the potential new AD metabolite was the main obstacle for its chemical structure elucidation. In this study, to increase the concentration of targeted metabolites, the influence of cellular morphology on AD metabolism in strain NJES-13^T was determined using glass bead-enhanced fermentation. Based on the cellular ultra-structural observation driven by bacterial EPSs, and quantitative analysis of the targeted metabolites, the successful increasing of the productivity of three AD metabolites was achieved. Afterward, a new frigocyclinone analogue was isolated and then identified as 2-hydroxy-frigocyclinone, as well as two other known ADs named 2-hydroxy-tetrangomycin (2-HT) and gephyromycin (GPM). Three AD metabolites were found to demonstrate different bioactivities. Both C-2 hydroxyl substitutes, 2-hydroxy-tetrangomycin and 2-hydroxy-frigocyclinone, exhibited variable inhibitory activities against Staphylococcus aureus, Bacillus subtilis and Candida albicans. Moreover, the newly identified 2-hydroxy-frigocyclinone also showed significant cytotoxicity against three tested human-derived cancerous cell lines (HL-60, Bel-7402 and A549), with all obtained IC₅₀ values less than 10 µM. Based on the genetic analysis after genomic mining, the plausible biogenetic pathway of the three bioactive ADs in strain NJES-13^T was also proposed.     

Enzyme Bioprospection of Marine-Derived Actinobacteria from the Chilean Coast and New Insight in the Mechanism of Keratin Degradation in Streptomyces sp. G11C

Marine actinobacteria are viewed as a promising source of enzymes with potential technological applications. They contribute to the turnover of complex biopolymers, such as pectin, lignocellulose, chitin, and keratin, being able to secrete a wide variety of extracellular enzymes. Among these, keratinases are a valuable alternative for recycling keratin-rich waste, which is generated in large quantities by the poultry industry. In this work, we explored the biocatalytic potential of 75 marine-derived actinobacterial strains, focusing mainly on the search for keratinases. A major part of the strains secreted industrially important enzymes, such as proteases, lipases, cellulases, amylases, and keratinases. Among these, we identified two streptomycete strains that presented great potential for recycling keratin wastes—Streptomyces sp. CHA1 and Streptomyces sp. G11C. Substrate concentration, incubation temperature, and, to a lesser extent, inoculum size were found to be important parameters that influenced the production of keratinolytic enzymes in both strains. In addition, proteomic analysis of culture broths from Streptomyces sp. G11C on turkey feathers showed a high abundance and diversity of peptidases, belonging mainly to the serine and metallo-superfamilies. Two proteases from families S08 and M06 were highly expressed. These results contributed to elucidate the mechanism of keratin degradation mediated by streptomycetes.     

西岛内港沉积物放线菌的分离及其抗菌活性研究

从西岛内港采集沉积物,采用自然风干后,湿热60℃、处理10 min的预处理方法,稀释涂布于GA培养基上,成功地分离得到44株海洋源的放线菌。经初步形态显微观察判断,其中18株为链霉菌,26株为非链霉菌,链霉菌中以小单孢菌为主。18株链霉菌抗香蕉枯萎病测试结果显示,菌株090314表现出一定的抗香蕉枯萎病菌(4号生理小种,FOC 4)活性。     

公海深海底层渔业国际管理进展

以底拖网为代表的深海底层渔业对深海脆弱海洋生态系统的危害受到国际社会的热切关注。2003年以来联合国大会多次通过决议,呼吁各国各自并通过RFMO/As采取行动,根据预防性原则,采用基于生态系统的管理方法,评估深海底层渔业对脆弱海洋生态系统的影响,若评估表明确有重大不利影响,则应采取有效措施限制深海底层渔业以降低这种影响;FAO主要从技术角度制定了《公海深海渔业管理国际指南》,为管理公海深海渔业和保护脆弱海洋生态系统提出了技术标准和管理框架;RFMO承担着具体执行深海底层渔业管理措施和监督管理的责任,在北大西洋、地中海、南太平洋的公海和南极水域,相关RFMO已采取了暂停部分区域底拖网渔业活动、收集数据、评估底拖网对脆弱海洋生态系统的影响等措施,在北太平洋,新成立的北太平洋渔业管理委员会将公海底层渔业管理作为首要目标。环保非政府组织和部分科学家呼吁禁止公海深海底层渔业,但各国对此的立场尚不一致,产业界大多持反对立场。近期来看,尚难以全面禁止公海的深海底层渔业。中国正在发展公海大洋渔业,需对此密切关注,加强跟踪研究以支撑决策,并应发展和使用选择性渔具和对生态环境无害的作业方式,防止对脆弱海洋生态系统产生损害性影响。     

Two New Cytotoxic Indole Alkaloids from a Deep-Sea Sediment Derived Metagenomic Clone

Two new indole alkaloids, metagenetriindole A (1) and metagenebiindole A (2), were identified from deep-sea sediment metagenomic clone derived Escherichia coli fermentation broth. The structures of new compounds were elucidated by spectroscopic methods. The two new indole alkaloids demonstrated moderately cytotoxic activity against CNE2, Bel7402 and HT1080 cancer cell lines in vitro.     

新疆硝尔库勒盐湖放线菌分离方法的初步研究

为了研究新疆硝尔库勒盐湖中的放线菌多样性,以微生物抗高渗透压的生理机制为指导,结合盐湖高盐、寡营养的特点,设计了含有在嗜盐微生物细胞中起渗透调节作用的多种碳源和氮源的分离培养基,以TP培养基、GW1培养基和F6培养基为对照,对新疆硝尔库勒湖沉积物样品进行分离。结果表明,海藻糖-肌酸培养基(B3)分离效果最好,能分离到10个属16个不同种的放线菌;海藻糖-天冬氨酸培养基(B1)和海藻糖-谷氨酸培养基(B2)分离效果次之,均能分离到5个属7个不同种的放线菌,并有2个潜在的放线菌新物种被发现(菌株编号为41376、41293)。这说明以渗透调节物作为碳源和氮源是高盐环境放线菌分离比较理想的选择,为以后高盐环境微生物的分离培养提供了理论指导。     

Isolation,Characterization, and Bioactivity Evaluation of 3-((6-Methylpyrazin-2-yl)methyl)-1H-indole,a New Alkaloid from a Deep-Sea-Derived Actinomycete Serinicoccus profundi sp. nov

One new alkaloid, 3-((6-methylpyrazin-2-yl)methyl)-1H-indole (1) was obtained from the deep-sea actinomycete Serinicoccus profundi sp. nov., along with five known compounds (2–6). Their structures were determined on the basis of detailed analysis of the 1D and 2D NMR as well as MS data. The new indole alkaloid displayed weak antimicrobial activity against Staphylococcus aureus ATCC 25923 with an MIC value of 96 μg/mL. It showed no cytotoxicity on a normal human liver cell line (BEL7402) and a human liver tumor cell line (HL-7702).     

Chemical Constituents of the Deep-Sea-Derived Penicillium solitum

A systematic chemical investigation of the deep-sea-derived fungus Penicillium solitum MCCC 3A00215 resulted in the isolation of one novel polyketide (1), two new alkaloids (2 and 3), and 22 known (4–25) compounds. The structures of the new compounds were established mainly on the basis of exhaustive analysis of 1D and 2D NMR data. Viridicatol (13) displayed moderate anti-tumor activities against PANC-1, Hela, and A549 cells with IC₅₀ values of around 20 μM. Moreover, 13 displayed potent in vitro anti-food allergic activity with an IC₅₀ value of 13 μM, compared to that of 92 μM for the positive control, loratadine, while indole-3-acetic acid methyl ester (9) and penicopeptide A (10) showed moderate effects (IC₅₀ = 50 and 58 μM, respectively).