Nicotinil Dibuthyl Glutamate Ester (NDBGE) is a synthetic compound that is an analog of antibiotic UK 3A from mycelium of Streptomyces sp.517-02. This compound was estimated to have anticancer activity similar to UK-3A. The aim of this research was to know the NDBGEs apoptotic inducing potency to human breast cancer T47D in vitro. DMEM + 1% PS + 10% FBS was added to human breast cancer T47D cells until cell concentration reached 1-5x/ 05eells/ml. NDBGE then was diluted by DMSO until 100ppm concentration. Propidium iodide with 50pg/m1 concentration was added to the T47D cells. The analysis was conducted by flow cytometry method. The result was that the apoptosis percentage of the cells was 12,35%.

Abstract Sea cucumber Holothuria scabra is a potential export commodity in the fisheries sub-sector. Some are used indirectly in extract form. Selection of the extraction method is carried out by considering its activity. One of the benefits that is often claimed in sea cucumber products is antimicrobial activity. This study aims to determine the antimicrobial activity of sea cucumbers after the extraction process using the reflux method. The ethanol 70% was used as solvent with citric acid as catalyst. Selection of the extraction solvent and catalyst is carried out by considering its safety for human consumption. The catalyst is used to hydrolyze polysaccharides. With the presence of a catalyst combine with various contact times, and temperature we try to assay its microbial activity. We used a fixed amount of solvent and the catalyst concentration was set from 0 to 1% (w/v), contact time 6 to 18 hours, temperature 25 to 55 °C. The experiment design was using factorial analysis at two levels. All the solution was filtered and evaporated to get the dried extract and freeze-dried before it was tested in S. aureus, B. subtilis, and E. coli to see its inhibition zone. The result showed that on SA catalyst and temperature had a significant effect on the inhibition zone. However, on BS and EC, only the catalyst has a significant effect on the inhibition zone. The best-obtained result was estimated using higher catalyst concentration, shorter extraction times, and the lowest temperature.

Current molecular PCR-based methods for identifying Streptococcus pneumoniae, the primary cause of pneumonia, meningitis, and other invasive diseases, are accurate but require expensive infrastructure and have a long run time, which restricts their use, particularly in developing countries. LAMP, or loop-mediated isothermal amplification, is a low-cost alternative to polymerase chain reaction (PCR) that can be used quickly because the reaction only takes place at a constant temperature. We aimed to develop a LAMP assay for rapid detection of Streptococcus pneumoniae serotypes in nasopharynx swab samples. The LAMP primers were designed using a conserved region of the lytA gene. An incubation time range of 30 to 60 minutes was studied to optimize the LAMP reaction. The real-time fluorescence intensity was monitored during the amplification reaction. Clinical nasopharynx swab samples identified as Streptococcus pneumoniae and their serotypes were tested to evaluate the performance of LAMP. To investigate the specificity of the LAMP, Streptococcus species samples and non-Streptococcus species samples were analyzed. In conclusion, the optimized LAMP assay is capable of detecting Streptococcus pneumoniae and its serotypes in nasopharynx swab samples.

The current study introduces a one-pot technique for synthesizing an environmentally benign and cheap composite adsorbent, namely ZnO-PPAC, for the adsorption of Pb(II). The designated adsorbent was prepared by incorporating green synthesized zinc oxide (ZnO) nanoparticles on activated carbon-derived from pineapple peel. The prepared adsorbents were characterized using XRD, SEM, EDS, FTIR, and BET techniques. The XRD pattern verifies that the ZnO was successfully synthesized and immobilized onto the PPAC in a one pot synthesis system. The surface areas of ZnOPPAC and PPAC adsorbents were 13.62 m2/g and 961.96 m2/g, respectively. The FTIR evaluation of the ZnO-PPAC adsorbent revealed several characteristic absorption peaks corresponding to -OH groups, C-O groups, C=C groups, C-N groups, and M-O groups. It was revealed that the adsorption of Pb(II) on the ZnO/PPAC adsorbent would not require any pH adjustment. The adsorption kinetics demonstrated that the adsorption of Pb(II) ions on PPAC and ZnO-PPAC better fitted pseudo-second-order kinetics (R2 = 0.999, both for PPAC and ZnO/PPAC) and followed the Freundlich model (R2 = 0.989 and 0.987 for PPAC and ZnO/PPAC adsorbent). According to the Freundlich model, the adsorption of Pb(II) ions onto the designated adsorbents involves a multilayer process. The maximum adsorption capacities of ZnO/PPAC and PPAC were calculated as 769 mg/g and 667 mg/g, respectively. Thermodynamic analysis indicated an exothermic and spontaneous nature, as suggested by the negative values of ΔHº and ΔGº. In summary, both the prepared adsorbents greatly exhibited a high adsorption capacity of Pb(II) ions that can be used for environmental remediation.

Myxobacteria are Gram-negative bacteria renowned for their valuable production of secondary metabolites; however, their diverse protease enzymes remain less explored. In this investigation, a novel strain was isolated from mangrove soil that produces alkaline protease. Using 16S rRNA sequence analysis, the strain was identified as Myxococcus virescens species. The extracellular alkaline protease MvCP of strain S2-2 was produced using growth media containing agro-industrial by-products such as soybean meal and tofu dregs. MvCP exhibited optimum activity within a pH range from 9 to 10 and temperatures ranging from 30 °C to 50 °C. MvCP activity was improved with Ca2⁺, Mg2⁺, and Zn2⁺ ions, whereas EDTA and β-mercaptoethanol at concentrations of 0.5 and 1.25 mM had no significant effect. MvCP was stable in chloroform, acetone, Triton X-100, Tween-20, and Tween 80. MvCP can be considered an eco-friendly option for use in household cleaning products and chitin extraction processes.

Citronella oil (CTO) is extracted from citronella leaves by maceration or steam distillation process, which has antibacterial and insect-repellent activities. However, the use of CTO is limited and requires modification in other formulations, such as microemulsion (ME), to increase its bioactivities. ME consists of oil, water, surfactant and/or cosurfactant and is commonly applied in food and beverages, cosmetics, and carrier for drug delivery applications. CTO was used as the oil phase for ME with nonionic surfactant and ethanol as a cosurfactant for lowering interfacial tension between oil and water phase. Subsequent observations regarding stability and antibacterial tests were carried out on ME formulations with surfactant/cosurfactant mixture of 2 due to its largest ME area. A hydrodynamic diameter analysis was also carried out to see the stability of the ME within a period of 50 d. ME with 10% CTO, 30% surfactant mixture, and 60% water showed the best formulation observed from the consistent hydrodynamic diameter measurement. In addition, ME with different formulations could inhibit the growth of Escherichia coli and Staphylococcus aureus by more than 90%. From this research, CTO-based ME potentially improve and develop drug carrier applications, especially via topical route.

The Gram-positive bacteria Streptomyces davaonensis and Streptomyces cinnabarinus have been the only organisms known to produce roseoflavin, a riboflavin (vitamin B 2 ) derived red antibiotic. Using a selective growth medium and a phenotypic screening, we were able to isolate a novel roseoflavin producer from a German soil sample. The isolation procedure was repeated twice, that is, the same strain could be isolated from the same location in Berlin 6 months and 12 months after its first isolation. Whole genome sequencing of the novel roseoflavin producer revealed an unusual chromosomal arrangement and the deposited genome sequence of the new isolate (G + C content of 71.47%) contains 897 genes per inverted terminal repeat, 6190 genes in the core and 107 genes located on an illegitimate terminal end. We identified the roseoflavin biosynthetic genes rosA , rosB and rosC and an unusually high number of riboflavin biosynthetic genes. Overexpression of rosA , rosB and rosC in Escherichia coli and enzyme assays confirmed their predicted functions in roseoflavin biosynthesis. A full taxonomic analysis revealed that the isolate represents a previously unknown Streptomyces species and we propose the name Streptomyces berlinensis sp. nov. for this roseoflavin producer.

Snail ( Achatina fulica ) is one of the sources of duck fodder commonly used by local breeders to meet protein needs of poultry farms. However, its shells as a by-product are rarely used and just become waste. To overcome this problem, we try to utilize the waste to be used as a source of bioceramic. Snail shells contain a lot of calcium which can be used as a calcium precursor to make bioceramics. One of the bioceramics that has the potential are tricalcium phosphate (TCP) and calcium pyrophosphate (CPP). They are mostly used as material for bone regeneration. It has advantages in terms of biocompatibility and osteoinductivity. In this study, we synthesized and characterized calcium phosphate from snails by coprecipitation method. It was then analyzed using X-Ray fluorescence (XRF), X-Ray diffraction (XRD), fourier transform infrared (FTIR), energy dispersive X-Ray (EDX), and scanning electron microscope (SEM) analysis to see its characteristic. Based on XRF analysis, snail shell (raw material) contains 81.83% of calcium and calcium phosphate mixed bioceramics (TCP and CPP). Average crystal size is 17.00 nm. The surface shape of the formed particles was irregular, and also flux, sphere, and fracture shaped. Nevertheless, interfering compounds were still found, hence a further purification process needs to be conducted.

A facile and eco-friendly procedure was developed to fabricate zinc oxide nanoparticles (ZnO NPs) using an aqueous extract of mango fruit peel (MFP), a by-product of agroindustry. The ZnO NPs were fabricated using zinc acetate as a precursor and MFP extract as a reducing and capping agent in a neutral environment (pH 7). The UV-visible spectrum supported the formation of ZnO NPs, showing a distinctive absorption peak at 368 nm. The presence of a ZnO crystalline phase was identified by X-ray diffraction (XRD) analysis. The Fourier transform infrared (FTIR) evaluation demonstrated that the biomolecules present in the MFP extract actively contributed to zinc ion reduction. According to the scanning electron microscopy (SEM) image, the surface morphology of ZnO NPs showed a mixture of spherical and flake-like shapes with particle sizes ranging from ~20 to ~90 nm. Based on antibacterial analysis using the agar diffusion method, the biosynthesized ZnO NPs at 3% w/v were active against Escherichia coli, Bacillus subtilis , and Staphylococcus aureus with diameter inhibitions of 8, 19, and 10 mm, respectively. In summary, this present work highlights that ZnO NPs can be synthesized using a by-product of agroindustry. More importantly, the biosynthesized ZnO NPs can be applied as an antibacterial agent.

Actinobacteria are mainly found in soil and some of them have properties that are common to bacteria and fungi, despite having quite distinct characteristics. Unlike bacterial colonies in general, which are clearly slimy and grow rapidly, some groups of actinobacteria colonies grow slowly by showing a powdery consistency and are tightly attached to the agar surface. Observations on a colony under the microscope showed that many of them form asexual spores for their reproduction. Many metabolite compounds generated by actinobacteria have promising activities like antioxidant and antagonistic activity against bacteria and fungi. The production of these compounds depends not only on the strain of the organism but also on the medium in which it is grown and the growth conditions. Moreover, agricultural by-products such as soybean meal are known to have high protein content, thus it can be potentially used as an alternative media for actinobacteria. In this research, ten actinobacterial strains were isolated from the soil. After seven days of cultivation with the medium containing soybean meal, the cultures were subjected to ethyl acetate extraction. Five extracts exhibited antibacterial properties against Bacillus subtilis with a zone of inhibition ranging from 10–14 mm. One extract could strongly inhibit Staphylococcus aureus with an inhibition zone of 21 mm. However, none of them were active against Escherichia coli . Five extracts demonstrated antioxidant DPPH radical scavenging activity with more than 40%.

Pineapple peel waste contains antioxidant molecules. This paper describes a facile, efficient, and environmentally friendly approach for synthesizing ZnO nanoparticles using pineapple peel aqueous extract. This method aims to reduce the use of harmful chemicals in producing ZnO nanoparticles using zinc nitrate as a salt precursor and evaluating their antibacterial. Pineapple peel aqueous extract was obtained by macerating pineapple peel powder in distilled water. Engaging Zn(NO 3 ) 2 , pineapple peel aqueous extract, and NaOH, ZnO NPs were successfully prepared by biosynthesis. X-ray diffraction (XRD) analysis and Fourier transform infrared (FTIR) spectroscopy was utilized to confirm the formation of ZnO NPs, while scanning electron microscopy (SEM) was employed to see the morphology and size distribution of nanoparticles. The antibacterial activity of pineapple peel extract-ZnO nanoparticles (pp-ZnO NPs) against Gram-positive and Gram-negative bacteria was evaluated by an agar disc diffusion method. The SEM images showed that by applying pineapple peel aqueous extract, the pp-ZnO with an average size below 100 nm with a mixture of flakes and rod-like shapes were obtained. The XRD pattern confirmed the wurtzite phase of ZnO, while antibacterial assay results showed that the pp-ZnO actively inhibited B. subtilis and E.coli . at concentrations of 5 and 10 wt%. Therefore, phyto-assisted synthesis mediated by pineapple peel wastes is a promising green approach to preparing ZnO nanoparticles.

Protease is a type of enzyme that hydrolyzes proteins into peptides and amino acids. The demand for protease for green technology in some industries like detergent, food, and leather is increasing nowadays. Some soil bacteria, especially the Bacillus strains, are known to have the ability to produce protease. Compared to other sources like plant and animal origin, microbial protease has more advantages as it can be produced at an industrial scale, short cultivation time, and are easy to harvest. This research aimed to screen the proteolytic bacteria from the bulk soil samples collected under mango trees and to study the optimum condition for protease production using the response surface methodology (RSM). Three bacterial strains (SH2CR, SH3CR, and SC4CR) were isolated and shown to have proteolytic activity. Based on 16S rRNA gene analysis, the strain SH2CR was close to Priestia megaterium NBRC 15308 T (96.97% similarity), while the strains SH3CR and SC4CR were related to Bacillus zanthoxyli strain 1433 T (100% similarity). One of them, SH2CR, was further studied using a fermenter at a one-liter production scale with the medium containing skim milk at 30°C. The best condition for protease production by SH2CR was achieved at 48 h incubation time, 300 rpm of agitation, and 1.25% skim milk.

Sponges and their associated microbial communities are known to produce chemically diverse metabolites that in nature serve as functional compounds required by the host. These compounds were also found to carry therapeutic importance as antimicrobial, anticancer and antioxidant agents. Actinomycetes are well known for their capability to produce many biologically active compounds, especially those from the genus Streptomyces. Due to the high demand for novel antibacterial drugs to combat antibiotic-resistant bacteria, finding novel bioactive compounds produced by actinomycetes that are isolated from extreme or underexplored habitats, is of importance and may pave the way to novel drug discovery. Novel or rare species of species of actinomycetes from marine sponges with altered biosynthetic pathways to adapt such environments might also provide novel scaffold molecules for drugs. Therefore, isolation of bioactive compounds from sponge symbiotic actinomycetes and getting information from their genomes on their biosynthetic genes are of importance. The chapter thus provides an overview on the sponge symbiotic and bioactive actinomycetes with examples from Atlantic and Pacific Oceans.

A novel myxobacterial strain ZKHCc1 1396T was isolated in 2017 from a soil sample collected along Chalus Road connecting Tehran and Mazandaran, Iran. It was a Gram-negative, rod-shaped bacterial strain that displayed the general features of Corallococcus, including gliding and fruiting body formation on agar and microbial lytic activity. Strain ZKHCc1 1396T was characterized as an aerobic, mesophilic, and chemoheterotrophic bacterium resistant to many antibiotics. The major cellular fatty acids were branched-chain iso-C17:0 2-OH, iso-C15:0, iso-C17:1, and iso-C17:0. The strain showed the highest 16S rRNA gene sequence similarity to Corallococcusterminator CA054AT (99.67%) and C. praedator CA031BT (99.17%), and formed a novel branch both in the 16S rRNA gene sequence and phylogenomic tree. The genome size was 9,437,609 bp, with a DNA G + C content of 69.8 mol%. The strain had an average nucleotide identity (ANI) value lower than the species cut-off (95%), and with the digital DNA–DNA hybridization (dDDH) below the 70% threshold compared to the closest type strains. Secondary metabolite and biosynthetic gene cluster analyses revealed the strain’s potential to produce novel compounds. Based on polyphasic taxonomic characterization, we propose that strain ZKHCc1 1396T represents a novel species, Corallococcus soli sp. nov. (NCCB 100659T = CIP 111634T).

A marine Alphaproteobacterium designated as strain NZ-96T was isolated in February 2021, from a sponge species (Demospongiae) collected in muddy sediments with boulders and old chimneys in Otago/Canterbury Slope, Pacific Ocean, New Zealand. The isolate was found to be Gram-negative, rod-shaped, aerobic, motile, and produced yellow-colored colonies. The isolate was positive for alkaline phosphatase, leucine arylamidase, trypsin, catalase, and oxidase and negative for α-galactosidase and urease. It was resistant to many antibiotics including hygromycin, trimethoprim, spectinomycin, ampicillin, oxytetracycline, cephalosporin, bacitracin, and polymyxin. The 16S rRNA gene-based phylogenetic analyses exhibited that strain NZ-96T belonged to the genus Qipengyuania and showed 98.3–98.8% 16S rRNA gene sequence similarity to its closest relatives. The major respiratory quinone was ubiquinone-10 (Q-10). The polar lipid profile consisted of phosphatidylcholine, sphingoglycolipid, phosphatidylglycerol, one unknown polar lipid, and three unknown glycolipids. The major fatty acids were C18:1ω12t, C16:0, C16:1ω7c, C17:1ω6c, C16:02-OH, and C14:0 2-OH. Carotenoid were produced. The crude extract showed pronounced activity against Staphylococcus aureus Newman and Bacillus subtilis DSM 10. Pairwise ANI and dDDH values of strain NZ-96T and closely related phylogenetic hits were below the threshold values of 95% and 70%, respectively. Genes for trehalose biosynthesis, aspartate-semialdehyde dehydrogenase, flagellar biosynthesis, fatty acid biosynthesis, and antibiotics resistance were present, which aids in isolate survival in a sea or ocean environment. The DNA G+C content was 60.8% (by genome). Based on data obtained by the polyphasic approach, strain NZ-96T (= DSM 112811T = NCCB 100842T) represents a novel species of the genus Qipengyuania, for which the name Qipengyuania pacifica sp. nov. is proposed.

Cream colored bacteria from marine agar, strain WH24, WH77, and WH80 were isolated from the gill of the Crassostrea gigas a Pacific oyster with a filter-feeding habit that compels accompanying bacteria to demonstrate a high metabolic capacity, has proven able to colonize locations with changing circumstances. Based on the 16S rRNA gene sequence, all strains had high similarity to Photobacterium arenosum CAU 1568T (99.72%). This study involved phenotypic traits, phylogenetic analysis, antimicrobial activity evaluation, genome mining, Co-cultivation experiments, and chemical studies of crude extracts using HPLC and LC-HRESIMS. Photobacterium arenosum WH24 and Zooshikella harenae WH53Twere co-cultivated for 3 days in a rotary shaker at 160 rpm at 30 °C, and LC-MS monitored the chemical profiles of the co-cultures on the third day. The UV chromatograms of the extracts of the co-cultivation experiments show that Zooshikella harenae WH53T could be inhibited by strain WH24. The high virulence of Photobacterium arenosum WH24 was confirmed by genome analysis. Gene groups with high virulence potential were detected: tssA (ImpA), tssB (ImpB/vipA), tssC (ImpC/vipB), tssE, tssF (ImpG/vasA), tssG (ImpH/vasB), tssM (IcmF/vasK), tssJ (vasD), tssK (ImpJ/vasE), tssL (ImpK/vasF), clpV (tssH), vasH, hcp, lapP, plpD, and tpsB family.

Three new bacterial strains, WHY3T, WH131T, and WH158T, were isolated and described from the hemolymph of the Pacific oyster Crassostrea gigas utilizing polyphasic taxonomic techniques. The 16S rRNA gene sequence analysis revealed that strain WHY3T was a member of the genus Winogradskyella, whereas strains WHI31T and WH158T were members of the genus Erythrobacter. According to the polygenomic study the three strains formed individual lineages with strong bootstrap support. The comparison of dDDH-and ANI values, percentage of conserved proteins (POCP), and average amino acid identity (AAl) between the three strains and their relatives established that the three strains represented two separate genera. Menaquinone-6 was reported as the major respiratory quinone in strain WHY3T and Ubiquinone-10 for strains WH131T and WH158T, respectively. The major cellular fatty acids for strain WHY3T were C15:0, anteiso-C15:1 ω7c, iso-C15:0, C16:1ω7c. The major cellular fatty acids for strains WH131T and WH158T were C14:02-OH and t18:1ω12 for WH131T and C17:0, and C18:1ω7c for strain WH158T. Positive Sudan Black B staining Indicated the presence of polyhydroxyalkanoic acid granules for strains WH131T and WH158T but not for strain WHY3T. The DNA G + C contents of strains WHY3T, WH131T and WH158T were 34.4, 59.7 and 56.6%, respectively. Gene clusters predicted some important genes involved in the bioremediation process. Due to the accomplishment of polyphasic taxonomy, we propose three novel species Winogradskyella luteola sp.nov. (type strain WHY3T = DSM 111804T = NCCB 100833T), Erythrobacter ani sp.nov. (WH131T = DSM 112099T = NCCB 100824T) and Erythrobacter crassostrea sp.nov. (WH158T = DSM 112102T = NCCB 100877T).

AIMS:Explore the diversity of culturable actinobacteria isolated from the Pacific oyster Crassostrea gigas with special emphasis on their antimicrobial activity.METHODS AND RESULTS:For the characterization of the isolated actinobacteria, a polyphasic approach was adopted and thereby phenotypic descriptions, phylogenetic analysis, evaluations of antimicrobial activities and chemical analyses of crude extracts through HPLC and LC-HRESIMS were performed. Five strains were isolated from C. gigas. The 16S rRNA gene analysis revealed that three of them were taxonomically affiliated to the genus Streptomyces and the other two strains were related to Micromonospora. High inhibition was detected against different test microorganisms such as Candida albicans, Staphylococcus aureus, Bacillus subtilis and Mycobacterium smegmatis. On the basis of the chemical analysis, 11 compounds from the active fractions of the crude extracts were determined, and 8 were related putatively to previously reported compounds.CONCLUSIONS:Actinobacteria isolated from C. gigas represent an interesting reservoir of antimicrobial compounds, and further study to uncover the full capacity of this source is encouraged.SIGNIFICANCE AND IMPACT:At present, the study of actinobacteria and their antimicrobial potential from uncommon sources as C. gigas is vital to the development of new therapeutic agents to cope with the widespread resistance of human pathogens.

A yellow bacterium from marine agar, strain WHA3T, was isolated from the mantel of the Pacific oysters Crassostrea gigas in the Wilhelmshaven Sea in northern Germany. Based on the 16S rRNA gene sequence, strain WHA3T had a high similarity to Pacificimonas flava JLT2015T (95.80%) and 94.79% to Pacificimonas aurantium JLT2012T. Furthermore, the dDDH and ANI value analysis between WHA3T and other closest type strains were lower than 70% and 95%, respectively. The percentage of conserved proteins (POCP) and the average amino acid identity (AAI) value against Pacificimonas flava JLT2015T and Pacificimonas aurantium JLT2012T represented in the ranges of higher than 50% and 60%, respectively. Strain WHA3T contained ubiquinone-10 (Q-10) as the predominant quinone, and the major fatty acids were C16:1 ω7c and C18:1 ω7c. Granules of polyhydroxyalkanoates (PHAs) were absent. The main polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, several sphingoglycolipids, an unknown phospholipid, an unknown glycolipid, and an unknown polar lipid. The polyamines contained spermidine and spermine. The DNA G+C content of strain WHA3T was 61.69%. An analysis of the whole-genome sequence in the frame of genome mining strain WHA3T predicted the presence of genomes for one-carbon metabolism, TonB-dependent transporters, vitamin B12 transporter, iron siderophore receptor protein, and other genes, some of which play important roles against restricted nutrient sources. The extract of strain WHA3T moderately inhibited the growth of Candida albicans DSM 1665. The polyphasic taxonomic analysis results suggested that strain WHA3T could be separated from its closest type strains. Strain WHA3T represents a novel species in the genus Pacificimonas, for which we propose the name Pacificimonas pallium sp. nov., with the type strain WHA3T (= DSM 111825T = NCCB 100832 T).