Comparative performance evaluation of chitosan based polymeric microspheres and nanoparticles as delivery system for bacterial ß-carotene derived from Planococcus sp. TRC1.

ß-carotene is a natural compound with immense healthcare benefits. To overcome insolubility and lack of stability which restricts its application, in this study, ß-carotene from Planococcus sp. TRC1 was entrapped into formulations of chitosan­sodium alginate microspheres (MF1, MF2 and MF3) and chitosan nanoparticles (NF1, NF2 and NF3). The maximum entrapment efficiency (%) and loading capacity (%) were 80.6 ± 4.28 and 26 ± 3.05 (MF2) and 92.1 ± 3.44 and 41.86 ± 4.65 (NF2) respectively. Korsmeyer-Peppas model showed best fit with release, revealing non-Fickian diffusion. Thermal and UV treatment exhibited higher activation energy (kJ/mol), 17.76 and 15.57 (MF2) and 37.03 and 19.33 (NF2) compared to free ß-carotene (3.7 and 3.9), uncovering enhanced stability. MF2 and NF2 revealed swelling index (%) 721 ± 1.7 and 18.1 ± 1.5 (pH 6.8) and particle size 69.5 ± 3.2 µm and 92 ± 2.5 nm respectively. FESEM, FT-IR, XRD and DSC depicted spherical morphology, intactness of functional groups and masking of crystallinity. The IC50 (µg ml-1) values for antioxidant and anticancer (A-549) activities were 33.1 ± 1.7, 45.1 ± 2.8, 39.3 ± 2.9 and 31.3 ± 1.7, 27.9 ± 2.4, 25.3 ± 2.2 for ß-carotene, MF2 and NF2 respectively with no significant cytotoxicity on HEK-293 cells and RBCs (p > 0.05). This comparative study of microspheres and nanoparticles may allow the diverse applications of an unconventional bacterial ß-carotene with promising stability and efficacies.

Heterologous production of novel and rare C30-carotenoids using Planococcus carotenoid biosynthesis genes.

BACKGROUND: Members of the genus Planococcus have been revealed to utilize and degrade solvents such as aromatic hydrocarbons and alkanes, and likely to acquire tolerance to solvents. A yellow marine bacterium Planococcus maritimus strain iso-3 was isolated from an intertidal sediment that looked industrially polluted, from the Clyde estuary in the UK. This bacterium was found to produce a yellow acyclic carotenoid with a basic carbon 30 (C30) structure, which was determined to be methyl 5-glucosyl-5,6-dihydro-4,4'-diapolycopenoate. In the present study, we tried to isolate and identify genes involved in carotenoid biosynthesis from this marine bacterium, and to produce novel or rare C30-carotenoids with anti-oxidative activity in Escherichia coli by combinations of the isolated genes. RESULTS: A carotenoid biosynthesis gene cluster was found out through sequence analysis of the P. maritimus genomic DNA. This cluster consisted of seven carotenoid biosynthesis candidate genes (orf1-7). Then, we isolated the individual genes and analyzed the functions of these genes by expressing them in E. coli. The results indicated that orf2 and orf1 encoded 4,4'-diapophytoene synthase (CrtM) and 4,4'-diapophytoene desaturase (CrtNa), respectively. Furthermore, orf4 and orf5 were revealed to code for hydroxydiaponeurosporene desaturase (CrtNb) and glucosyltransferase (GT), respectively. By utilizing these carotenoid biosynthesis genes, we produced five intermediate C30-carotenoids. Their structural determination showed that two of them were novel compounds, 5-hydroxy-5,6-dihydro-4,4'-diaponeurosporene and 5-glucosyl-5,6-dihydro-4,4'-diapolycopene, and that one rare carotenoid 5-hydroxy-5,6-dihydro-4,4'-diapolycopene is included there. Moderate singlet oxygen-quenching activities were observed in the five C30-carotenoids including the two novel and one rare compounds. CONCLUSIONS: The carotenoid biosynthesis genes from P. maritimus strain iso-3, were isolated and functionally identified. Furthermore, we were able to produce two novel and one rare C30-carotenoids in E. coli, followed by positive evaluations of their singlet oxygen-quenching activities.

Bioprospecting potentials of moderately halophilic bacteria and the isolation of squalene producers from Kuwait sabkha.

Sabkhas in Kuwait are unique hypersaline marine environments under-explored for bacterial community composition and bioprospecting. The 16S rRNA sequence analysis of 46 isolates with distinct morphology from two Kuwait sabkhas recovered 11 genera. Phylum Firmicutes dominated these isolates, and Bacillus (32.6%) was recovered as the dominant genera, followed by Halococcus (17.4%). These isolates were moderately halophilic, and some of them showed tolerance and growth at extreme levels of salt (20%), pH (5 and/or 11), and temperature (55 °C). A higher percentage of isolates harbored protease (63.0), followed by DNase (41.3), amylase (41.3), and lipase (32.6). Selected isolates showed antimicrobial activity against E. faecalis and isolated Halomonas shengliensis, and Idiomarina piscisalsi harbored gene coding for dNDP-glucose 4,6-dehydratase (Glu 1), indicating their potential to produce biomolecules with deoxysugar moieties. Palmitic acid or oleic acid was the dominant fatty acid, and seven isolates had some polyunsaturated fatty acids (linolenic or γ-linolenic acid). Interestingly, six isolates belonging to Planococcus and Oceanobacillus genus produced squalene, a bioactive isoprenoid molecule. Their content increased 30-50% in the presence of Terbinafine. The potential bioactivities and extreme growth conditions make this untapped bacterial diversity a promising candidate for future bioprospecting studies.

Genomic insights into the salt tolerance and cold adaptation of Planococcus halotolerans SCU63T.

Planococcus halotolerans, recently described as a novel species with SCU63T as the type strain, is capable of thriving in up to 15% NaCl and temperatures as low as 0 °C. To better understand its adaptation strategies at the genomic level, strain SCU63T was subjected to whole-genome sequencing and data mining. The high-quality assembly yielded 17 scaffolds with a genome size of 3,622,698 bp. Its genome harbors 3683 protein-coding sequences and 127 RNA genes, as well as three biosynthetic gene clusters and 25 genomic islands. The phylogenomic tree provided compelling insights into the evolutionary relationships of Planococcus. Comparative genomic analysis revealed key similarities and differences in the functional gene categories among Planococcus species. Strain SCU63T was shown to have diverse stress response systems for high salt and cold habitats. Further comparison with three related species showed the presence of numerous unique gene clusters in the SCU63T genome. The strain might serve as a good model for using extremozymes in various biotechnological processes.

Highly enantioselective resolution of racemic 1-phenyl-1,2-ethanediol to (S)-1-phenyl-1,2-ethanediol by Kurthia gibsonii SC0312 in a biphasic system.

The asymmetric resolution of racemic 1-phenyl-1,2-ethanediol (PED) to (S)-PED by Kurthia gibsonii SC0312 (K. gibsonii SC0312) was conducted in a biphasic system comprised of an organic solvent and aqueous phosphate buffer. The impacts of organic solvents on the whole cell catalytic activity, metabolic activity, membrane integrity, and material distribution were first evaluated. The results showed that all organic solvents, except for dibutyl phthalate, showed a detrimental effect on the metabolic activity of the cells, especially for those with low log P values. All organic solvents were capable of changing the membrane permeability and membrane integrity of the cells. Moreover, some organic solvents showed a good extraction of the oxidation product. Finally, a high yield of 47.7 % of (S)-PED was obtained by the asymmetric resolution of racemic PED using K. gibsonii SC0312 in a biphasic system under the optimal conditions: racemic PED 120 mM, temperature 35 °C, reaction time 6 h, 180 rpm, and a volume ratio of dibutyl phthalate to aqueous phosphate buffer of 1:1. The optical purity of (S)-PED increased from 51.3 % to >99 %. This work described an efficient approach to improve reaction efficiency, and constructed a highly effective biphasic reaction system for the fabrication of (S)-PED via K. gibsonii SC0312.

Highly selective resolution of racemic 1-phenyl-1,2-ethanediol by a novel strain Kurthia gibsonii SC0312.

Chiral 1-phenyl-1,2-ethanediol (PED) performs vital effect for the preparation of pharmaceuticals, agrochemicals and cosmetics. In the study, a newly isolated strain Kurthia gibsoniiSC0312 with the ability to selectively oxidize racemic PED to achieve (S)-PED was evaluated in the aqueous reaction system. The strain showed excellent catalytic performances within the range of pH 5·5-8·5, temperature 25-45°C and the amount of cell 15 mg ml-1 to 30 mg ml-1 . Besides, 2-hydroxyacetophenone (HAP) as the oxidation product displayed a stronger inhibition to the catalytic activity of cell, only remaining <63% of catalytic activity after incubation at 40 mmol l-1 HAP for 6 h. For various metal ions, Cu2+ can obviously improve 1·7 times of the catalytic activity of cell at the concentration of 0·2 mmol l-1 . Acetone can stimulate the catalytic capacity of cell to improve the optical purity of (S)-PED at the PED concentration of 80 mmol l-1 , up to appropriately 94% from 85·4%; compared to the resting cell, growing cell exerted no positive effect in the yield and optical purity. Finally, a highly effective kinetic resolution system of racemic PED by the new strain was obtained, with the (S)-PED yield of 41% and optical purity of 94%. SIGNIFICANCE AND IMPACT OF THE STUDY: Biocatalyst is a vital component in the process of biotransformation. There are a growing number of studies of biocatalyst reporting the preparation of enantiomer of 1-phenyl-1,2-ethanediol. And the performance of this preparation reaction is also gradually improving. This study is the first to demonstrate that Kurthia gibsonii can efficiently and selectively oxidize racemic 1-phenyl-1,2-ethanediol, and we assess the effect of various factors on the catalytic performance of the strain. The work adds to a growing body of evidence for using biocatalytic method in the synthesis of chiral 1-phenyl-1,2-ethanediol and provides a probable approach to mine excellent properties of enzymes.

Combining chemical flocculation and bacterial co-culture of Cupriavidus taiwanensis and Ureibacillus thermosphaericus to detoxify a hardwood hemicelluloses hydrolysate and enable acetone-butanol-ethanol fermentation leading to butanol.

Butanol, a fuel with better characteristics than ethanol, can be produced via acetone-butanol-ethanol (ABE) fermentation using lignocellulosic biomass as a carbon source. However, many inhibitors present in the hydrolysate limit the yield of the fermentation process. In this work, a detoxification technology combining flocculation and biodetoxification within a bacterial co-culture composed of Ureibacillus thermosphaericus and Cupriavidus taiwanensis is presented for the first time. Co-culture-based strategies to detoxify filtered and unfiltered hydrolysates have been investigated. The best results of detoxification were obtained for a two-step approach combining flocculation to biodetoxification. This sequential process led to a final phenolic compounds concentration of 1.4 g/L, a value close to the minimum inhibitory level observed for flocculated hydrolysate (1.1 g/L). The generated hydrolysate was then fermented with Clostridium acetobutylicum ATCC 824 for 120 h. A final butanol production of 8 g/L was obtained, although the detoxified hydrolysate was diluted to reach 0.3 g/L of phenolics to ensure noninhibitory conditions. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2753, 2019.

Simplistic approach in extracellular synthesis of silver nanoparticles via bioreducing potential of Planococcus plakortidis strain BGCC-51 isolated from dye industry effluent soil.

Here, extracellular synthesis of silver nanoparticles (AgNPs) was carried out by Planococcus plakortidis strain BGCC-51 isolated from dye industry effluent soil. The microbes were isolated, screened, and characterised by molecular analysis (accession number KX776160). The optimisation of synthesis of AgNPs to determine the optimum substrate level (1-5 mM), pH (5-9), and temperature (25-55°C) were further carried out. P. plakortidis strain BGCC-51 gave best yield of AgNPs at substrate concentration 5 mM, pH 8, and at 35°C. Synthesised AgNPs were characterised by scanning electron microscope and high-resolution transmission electron microscope. The size of synthesised AgNPs was in the range of 20-40 nm having spherical morphology. The AgNPs were found to show antimicrobial activity against bacteria such as Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853), and Staphylococcus aureus (ATCC 29213).

Characterization of Solibacillus silvestris strain AM1 that produces amyloid bioemulsifier.

Solibacillus silvestris AM1 was the first strain from the genus to be reported for the production of a functional amyloid and its potential use as a surface active agent, a thermostable glycoprotein amyloid bioemulsifier BE-AM1 capable of influencing environment and biofilm formation. Phylogenetic analysis based on 16S rRNA gene, molecular characterization studies on the basis of DNA-DNA hybridization and chemotaxonomic fatty acid methyl ester (FAME) analysis showed that S. silvestris AM1 as a strain matches with the type strain S. silvestris HR3-23. But strain AM1 differs from the type strain HR3-23 in carbon substrate utilization studies along with amyloid bioemulsifier production ability with potential industrial and environmental applications. S. silvestris AM1 exhibited bioemulsifier production at wide range of factors like pH and NaCl concentrations, while temperature influenced the bioemulsifier production indirectly (since it affected the growth). Bioemulsifier production was observed even at oligotrophic conditions (0.5 mg ml-1 ) seen usually in its native environment. In this study, we have characterized the amyloid producing S. silvestris AM1 taxonomically and also analyzed 16S rDNA of 103 sequences of Solibacillus sp. available, which indicated the possibility of new species in this genus and can be studied for industrially and environmentally important biomolecules.

Kurthia ruminicola sp. nov., isolated from the rumen contents of a Holstein cow.

Gram-staining-positive, motile, rod-shaped bacteria, designated as H31022T and H31024 was isolated from rumen contents of a Holstein cow. Optimum growth occurred at 25°C and pH 7.0 on R2A agar medium. Oxidase and catalase activities are positive. The 16S rRNA gene sequence (1,452 bp) of the new isolates revealed they belong to the genus Kurthia of the phylum Firmicutes. Highest gene sequence similarities were assessed to be with Kurthia massiliensis JC30T (98.4%), Kurthia senegalensis JC8ET (97.5%), and Kurthia populi 10y-14T (97.4%). Kurthia sibirica DSM 4747T (97.3%), Kurthia zopfii NBRC 101529T (97.0%), and Kurthia gibsonii NCIMB 9758T (96.7%). DNA G + C content of strains H31022T and H31024 were 34.4% and 39.7%. Strains H31022T and H31024 has the following chemotaxonomic characteristics; the major fatty acids are iso-C15:0, iso-C14:0 and anteiso-C15; polar lipid profile contained diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE), unknown aminophospholipids (APL), unknown glycolipids (GL), unknown phospholipids (PL), and unknown polar lipids (L); the major quinone is MK-7. Based on polyphasic taxonomic analysis, strains H31022T (= KCTC 33923T = JCM 19640T) and H31024 (= KCTC 33924T = JCM 19641T) identified a novel species in the genus Kurthia for which the name Kurthia ruminicola sp. nov. is proposed.

Bioemulsifier (BE-AM1) produced by Solibacillus silvestris AM1 is a functional amyloid that modulates bacterial cell-surface properties.

A novel estuarine bacterial strain, Solibacillus silvestris AM1, produces an extracellular, thermostable and fibrous, glycoprotein bioemulsifier (BE-AM1). The amyloid nature of the bioemulsifier (BE-AM1) was confirmed by biophysical techniques (Congo red based polarization microscopy, ThioflavinS based fluorescent microscopy, fibrous arrangement in transmission electron microscopy and secondary structure measurement by FTIR and CD spectrum analysis). Cell-bound BE-AM1 production by S. silvestris AM1 during the mid-logarithmic phase of growth coincided with a decrease in cell surface hydrophobicity, and an increase in cell autoaggregation and biofilm formation. It was observed that the total interfacial interaction energy ([Formula: see text]) for the surface of the bioemulsifier producing S. silvestris AM1 and different derivatized surfaces of polystyrene (silanized and sulfonated) was found to support biofilm formation. This study has revealed that the BE-AM1, a bacterial bioemulsifier, is a functional amyloid and has a role in biofilm formation and cell surface modulation in S. silvestris AM1.

Characterization of the mechanism of prolonged adaptation to osmotic stress of Jeotgalibacillus malaysiensis via genome and transcriptome sequencing analyses.

Jeotgalibacillus malaysiensis, a moderate halophilic bacterium isolated from a pelagic area, can endure higher concentrations of sodium chloride (NaCl) than other Jeotgalibacillus type strains. In this study, we therefore chose to sequence and assemble the entire J. malaysiensis genome. This is the first report to provide a detailed analysis of the genomic features of J. malaysiensis, and to perform genetic comparisons between this microorganism and other halophiles. J. malaysiensis encodes a native megaplasmid (pJeoMA), which is greater than 600 kilobases in size, that is absent from other sequenced species of Jeotgalibacillus. Subsequently, RNA-Seq-based transcriptome analysis was utilised to examine adaptations of J. malaysiensis to osmotic stress. Specifically, the eggNOG (evolutionary genealogy of genes: Non-supervised Orthologous Groups) and KEGG (Kyoto Encyclopaedia of Genes and Genomes) databases were used to elucidate the overall effects of osmotic stress on the organism. Generally, saline stress significantly affected carbohydrate, energy, and amino acid metabolism, as well as fatty acid biosynthesis. Our findings also indicate that J. malaysiensis adopted a combination of approaches, including the uptake or synthesis of osmoprotectants, for surviving salt stress. Among these, proline synthesis appeared to be the preferred method for withstanding prolonged osmotic stress in J. malaysiensis.

Biocalcifying Bacillus subtilis cells effectively consolidate deteriorated Globigerina limestone.

Microbially induced calcite precipitation occurs naturally on ancient limestone surfaces in Maltese hypogea. We exploited this phenomenon and treated deteriorated limestone with biocalcifying bacteria. The limestone was subjected to various mechanical and physical tests to present a statistically robust data set to prove that treatment was indeed effective. Bacillus subtilis conferred uniform bioconsolidation to a depth of 30 mm. Drilling resistance values were similar to those obtained for freshly quarried limestone (9 N) and increased up to 15 N. Treatment resulted in a high resistance to salt deterioration and a slow rate of water absorption. The overall percentage porosity of treated limestone varied by ±6 %, thus the pore network was preserved. We report an eco-friendly treatment that closely resembles the mineral composition of limestone and that penetrates into the porous structure without affecting the limestones' natural properties. The treatment is of industrial relevance since it compares well with stone consolidants available commercially.

Rummeliibacillus suwonensis sp. nov., isolated from soil collected in a mountain area of South Korea.

A Gram-positive, facultatively aerobic, rod-shaped, non-motile, terminal spore-forming bacterium, designated strain G20(T), was isolated from soil collected in a mountain region of Suwon, South Korea. On the basis of 16S rRNA gene sequence similarity, this strain was shown to be related to Rummeliibacillus pycnus NBRC 101231(T) (97.4%) and Rummeliibacillus stabekisii KSC-SF6g(T) (95.7%). DNA-DNA hybridization studies showed 42% and 50% similarity of strain G20(T) with R. pycnus NBRC 101231(T) and R. stabekisii KSC-SF6g(T), respectively. The DNA G content of G20(T) was 37.8 mol%, the major cellular fatty acids were iso-C15:0 and anteiso-C15:0, and the predominant menaquinones were MK-7 and MK-8. On the basis of phylogenetic, chemotaxonomic, and phenotypic characteristics, we propose this strain to be a novel species and the third member of genus Rummeliibacillus. We suggest the name Rummeliibacillus suwonensis sp. nov. The type strain is G20(T) (KACC 17316(T) =KEMB 9005-003(T) =JCM 19065(T)).

Isolation of a novel strain of Planomicrobium chinense from diesel contaminated soil of tropical environment.

A novel bacterial strain (B6) degrading high concentration of diesel oil [up to 2.5% (v/v)] was isolated from a site contaminated with petroleum hydrocarbons in the state of Chhattisgarh, India. The strain demonstrated efficient degradation for diesel oil range alkanes (C14 to C36 i.e., mostly linear chain alkanes). It was identified to be 99% similar to Planomicrobium chinense on the basis of partial 16S rRNA gene sequencing and biochemical characteristics. The efficiency of degradation was optimized at pH 7.2 and temperature at 32 °C. GC analysis demonstrated complete mineralization of higher chain alkanes into lower chain alkanes within 96 h. The organism also displayed surface tension reduction by producing stable emulsification on the onset of stationary phase. A multidimensional characteristics of the strain to grow at a high temperature range, resistance to various heavy metals as well as tolerance to moderate concentration of NaCl makes it suitable for bioremediation of soil contaminated with diesel oil in tropical environment.

Chryseomicrobium imtechense gen. nov., sp. nov., a new member of the family Planococcaceae.

A Gram-stain-positive, rod-shaped, yellow, non-motile, non-spore-forming, strictly aerobic bacterial strain, designated MW 10(T), was isolated from seawater of the Bay of Bengal, India, and was subjected to a polyphasic taxonomic study. Analysis of the 16S rRNA gene sequence revealed that strain MW 10(T) showed highest similarity to the type strains of Psychrobacillus psychrodurans (96.15 %) and Psychrobacillus psychrotolerans (96.01 %) and showed less than 96 % similarity to members of the genera Paenisporosarcina, Planococcus, Sporosarcina and Planomicrobium. Phylogenetic analysis based on the 16S rRNA gene sequence showed that strain MW 10(T) formed a clade separate from members of closely related genera. The morphological, physiological and chemotaxonomic characteristics of strain MW 10(T) differed from those of members of closely related genera. The major fatty acid in strain MW 10(T) was iso-C(15 : 0) and the menaquinones were MK-7 (48.4 %), MK-8 (32.3 %), MK-7(H(2)) (13.7 %) and MK-6 (5.6 %). The polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, an unknown phospholipid, an unknown lipid and an unknown glycolipid. The cell-wall peptidoglycan type was l-Lys-d-Asp. The genomic DNA G+C content (53.4 mol%) of strain MW 10(T) was significantly different from those of members of closely related genera. On the basis of its morphological, physiological and chemotaxonomic characteristics as well as our phylogenetic analysis, we conclude that strain MW 10(T) is a member of a novel genus and species, for which the name Chryseomicrobium imtechense gen. nov., sp. nov., is proposed. The type strain of Chryseomicrobium imtechense is MW 10(T) ( = MTCC 10098(T)  = JCM 16573(T)).