Novel Alkaloids from Marine Actinobacteria: Discovery and Characterization

The marine environment is an excellent resource for natural products with therapeutic potential. Its microbial inhabitants, often associated with other marine organisms, are specialized in the synthesis of bioactive secondary metabolites. Similar to their terrestrial counterparts, marine Actinobacteria are a prevalent source of these natural products. Here, we discuss 77 newly discovered alkaloids produced by such marine Actinobacteria between 2017 and mid-2021, as well as the strategies employed in their elucidation. While 12 different classes of alkaloids were unraveled, indoles, diketopiperazines, glutarimides, indolizidines, and pyrroles were most dominant. Discoveries were mainly based on experimental approaches where microbial extracts were analyzed in relation to novel compounds. Although such experimental procedures have proven useful in the past, the methodologies need adaptations to limit the chance of compound rediscovery. On the other hand, genome mining provides a different angle for natural product discovery. While the technology is still relatively young compared to experimental screening, significant improvement has been made in recent years. Together with synthetic biology tools, both genome mining and extract screening provide excellent opportunities for continued drug discovery from marine Actinobacteria.

SECONDARY METABOLITES WITH ANTIBIOTIC ACTIVITY OF MARINE ACTINOBACTERIA

Marine actinobacteria are active producers and an unused rich source of various biologically active secondary metabolites, such as antibiotics, antitumor, antiviral and antiinflammatory compounds, biopesticides, plant growth hormones, pigments, enzymes, enzyme inhibitors.In this review describes data from current literature sources for the period from 2017 to 2021 about various bioactive compounds that produce marine actinobacteria, their antibiotic activity and biotechnological potential, the main groups of secondary metabolites and their producers.

Multifaceted bioproperties of Streptomyces bacillaris ANS2 isolated from Andaman and Nicobar Islands, India

The rich warehouse called marine actinobacteria has enticed researchers for several decades. The assessment of their metabolites has evidenced several diverse and multiple biological properties vacillating from antibacterial, antituberculosis, antioxidant, anticancer to mosquitocidal. Therefore, the present study has been aimed to reconnoitre the isolation and portrayal of the promising marine actinobacteria named Streptomyces bacillaris ANS2 isolated from the mangrove sediment collected from Andaman and Nicobar islands. The ethyl acetate extract of ANS2 showed broad spectrum activity against various pathogens including Escherichia coli (18.0±0.3 mm), Salmonella paratyphi (16.0±0.6 mm) and Klebsiella pneumonia (14.8±0.7 mm). It showed maximum activity against drug sensitive strain of Mycobacterium tuberculosis (98.2±0.36%) followed by M. tuberculosis MDR strain (97.04±1.32%) and M. tuberculosis H37Rv (91.80±0.45%). The antioxidant activity by DPPH assay of ANS2 showed 29.9±0.38 - 61.35±0.78 % free radical scavenging at 100μg/ml - 500μg/ml as compared to the standard ascorbic acid which showed 96±1.0% at 100μg/ml concentration. The anticancer activity showed 88.12±0.9% and 89.5±0.26% inhibition against HT 29 (colon cancer) and HeLa (cervical cancer) cell line at 1000μg/ml concentration. The maximum larval mortality was recorded on Culex quinquefasciatus(LC50 =1100.134 μg/ml and r2= 0.99) and Aedes aegypti (LC50 =690.620 μg/ml and r2= 0.99).The biochemical, morphological and 16S rRNA gene analysis study clearly confirmed that the promising strain belonged to the genus Streptomyces bacillaris and the phylogenetic tree confirmed it as S. bacillaris. Results showed that S.bacillaris ANS2 showed multidimensional bioproperties.

Isolation and Structure Elucidation of Desferrioxamine B and the New Desferrioxamine B2 Antibiotics from a Brown Marine Macroalga Carpodesmia tamariscifolia Associated Streptomyces Isolate

Today, emerging infectious diseases caused by multidrug-resistant bacteria (MDRB) are a major public health problem. These bacteria are gradually becoming more resistant to conventional antimicrobial agents. Thus, there is an urgent requirement to explore new antimicrobial compounds. This study focuses on a screening program of marine actinobacteria for useful bioactive compounds against MDRB, and four endophytic actinobacteria strain isolated from the unexploited marine brown alga Carpodesmia tamariscifolia, harvested from the Atlantic coast of Morocco, were screened for their antimicrobial activities using the agar diffusion assay. Fermentation broths of the two selected promising isolates KC179 and KC180 were extracted with different organic solvents and showed antibacterial activity against methicillin-resistant Staphylococcus aureus, Acinetobacter baumannii, and Pseudomonas aeruginosa. From the butanolic extract of KC180's culture broth and in addition to the known metabolite desferrioxamine B, a new desferrioxamine derivative, desferrioxamine B2, was purified using flash chromatography and reversed-phase HPLC, and its structure was elucidated using HRMS and NMR spectroscopy. The 16S rRNA molecular taxonomic characterization of the producing strain KC180 showed Streptomyces albidoflavus as the nearest relative, with a sequence similarity of 99.71 %.

Streptomyces tardus sp. nov.: A Slow-Growing Actinobacterium Producing Candicidin, Isolated From Sediments of the Trondheim Fjord

Marine environments are home to an extensive number of microorganisms, many of which remain unexplored for taxonomic novelty and functional capabilities. In this study, a slow-growing Streptomyces strain expressing unique genomic and phenotypic characteristics, P38-E01T, was described using a polyphasic taxonomic approach. This strain is part of a collection of over 8,000 marine Actinobacteria isolates collected in the Trondheim fjord of Norway by SINTEF Industry (Trondheim, Norway) and the Norwegian University of Science and Technology (NTNU, Trondheim, Norway). Strain P38-E01T was isolated from the sediments of the Trondheim fjord, and phylogenetic analyses affiliated this strain with the genus Streptomyces, but it was not closely affiliated with other described species. The closest related type strains were Streptomyces daliensis YIM 31724T (98.6%), Streptomyces rimosus subsp. rimosus ATCC 10970T (98.4%), and Streptomyces sclerotialus NRRL ISP-5269T (98.3%). Predominant fatty acids were C16:0 iso, C16:0, and Summed Feature 3, and the predominant respiratory quinones were MK-10(H6), MK-10(H4), and MK9(H4). The main polar lipids were identified as diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, and phosphoglycolipid. The whole-cell sugars were glucose, ribose, and in minor amounts, mannose. The cell wall peptidoglycan contained LL-diaminopimelic acid. The draft genome has a size of 6.16 Mb, with a %G + C content of 71.4% and is predicted to contain at least 19 biosynthetic gene clusters encoding diverse secondary metabolites. Strain P38-E01T was found to inhibit the growth of the pathogenic yeast Candida albicans ATCC 90028 and a number of Gram-positive bacterial human and plant pathogens. Metabolites extracted from cultures of P38-E01T were analyzed by mass spectrometry, and it was found that the isolate produced the antifungal compound candicidin. Phenotypic and chemotaxonomic signatures, along with phylogenetic analyses, distinguished isolate P38-E01T from its closest neighbors; thus, this isolate represents a novel species of the genus Streptomyces for which the name Streptomyces tardus sp. nov. (P38-E01T = CCM 9049T = DSM 111582T) is proposed.

Degradation and Detoxification of Congo Red azo dye by Immobilized Laccase of Streptomyces sviceus

The discharge of textile effluents enriched with reactive azo dyes is of critical importance owing to inability of the dyes to degrade in waste water and their carcinogenic, mutagenic effects to various organisms. This study initiated based on the need to gaze into molecular mechanism of marine bacterial bioremediation process to develop strategies for the decolorization and detoxification of the synthetic azo dyes. The experimental work carried out to explore decolorization and degradation efficacy of laccase derived from marine actinobacteria, Streptomyces sviceus by choosing Congo red-21 as model azo dye. The extracellular production of laccase was confirmed with plate assay in medium supplemented with ABTS as substrate. Laccase was purified to homogeneity from 72hrs culture of Streptomyces sviceus by Fast performance liquid chromatography and the molecular size of laccase was noticed as 60 kDa. The purified laccase was immobilized with an efficiency of 82% by Calcium alginate method. The crude, purified and immobilized forms of the laccase enzyme was used to decolorize the Congo red-21. Crude laccase enzyme showed 69% of decolorization of Congo red-21 after 48h where as purified and immobilized laccase represented 78% and 92% of colour removal after 24 h respectively. Fourier-transform infrared spectroscopy, High Performance Liquid Chromatography and Gas chromatography–mass spectrometry were used to unravel the molecular mechanism of dye detoxification and also identify nontoxic products released from Congo Red-21 upon administration with immobilized laccase. Based on GC-MS data, it may deduce that immobilized laccase of Streptomyces sviceus cleaves the Congo red-21 dye followed by oxidative cleavage, desulfonation, deamination, demethylation process.

Screening of Enzyme Producers with Keratinase Activity among Marine Actinobacteria

About 2 million tons of feathers are produced annually around the world as a by-product of poultry farming. Due to the lack of funds and the complexity of processing, they have become one of the main environmental pollutants. The biodegradation of feathers by keratinolytic microorganisms has proven to be an effective, environmentally friendly and cost-effective method of bioconverting feather waste into a nutritious, balanced and easily digestible product that contains free amino acids, peptides and ammonium ions. Aim. To investigate the ability of marine actinobacteria to synthesize enzymes with keratinolytic activity and to study some of the physicochemical properties of the most active enzyme preparation. The object of the study was 10 strains of actinobacteria isolated from bottom sediments in the area of the Pradneprovsky trench of the Black Sea shelf. Methods. Caseinolytic (general proteolytic) activity was determined by the Anson method modified by Petrova, based on the quantitative determination of tyrosine, which is formed during the enzymatic hydrolysis of casein. Keratinase activity was determined by UV absorption at 280 nm of the hydrolysis products of keratin-containing raw materials. The cultivation of actinobacteria was carried out in a liquid nutrient medium with the addition of defatted chicken feathers as the main source of carbon and nitrogen. Results. The ability to hydrolyze keratin was found in five cultures. Moreover, all the strains studied were practically unable to break down casein. The Acty 9 strain (12 U/ml) showed the highest keratinase activity. Additional introduction of NaCl to the nutrient medium did not have a positive effect on the enzymes synthesis. The study of the physicochemical properties of the enzyme preparation Acty 9 showed that the pH and thermooptimum were 9.0 and 60°C, respectively. It retained 100% of the initial activity in the range of pH 7.0–10.0 after 3 h and 95% activity at pH 8.0 after 24 h of incubation. The studied enzyme preparation was thermostable, since it remained active for 3 h at 50°C and 1 h at 60°C. Conclusions. The extracellular keratinase synthesized by actinobacterium Acty 9 is promising for further research, since the enzyme is pH and thermostable and is not inferior in its physicochemical properties to those previously described in the literature.