Amelioration of NaCl stress on germination, growth, and nitrogen fixation of Vicia faba at isosmotic Na-Ca combinations and Rhizobium.

MAIN CONCLUSION: The Na+/Ca2+ ratio of 1/5 ameliorated the inhibitory action of NaCl and improved the germination and growth of Vicia faba. Addition of Rhizobium also enhanced nodulation and nitrogen fixation. Casting light upon the impact of salinity stress on growth and nitrogen fixation of Vicia faba supplemented with Rhizobium has been traced in this work. How Ca2+ antagonizes Na+ toxicity and osmotic stress of NaCl was also targeted in isosmotic combinations of NaCl and CaCl2 having various Na+:Ca2+ ratios. Growth of Vicia faba (cultivar Giza 3) was studied at two stages: germination and seedling. At both experiments, seeds or seedlings were exposed to successively increasing salinity levels (0, 50, 100, 150, and 200 mM NaCl) as well as isosmotic combinations of NaCl and CaCl2 (Na+:Ca2+ of 1:1, 1:5, 1:10, 1:15, 1:18, and 1: 20), equivalent to 150 mM NaCl. Inocula of the local nitrogen-fixing bacteria, Rhizobium leguminosarum (OP715892) were supplemented at both stages. NaCl salinity exerted a negative impact on growth and metabolism of Vicia faba; inhibition was proportional with increasing salinity level up to the highest level of 200 mM. Seed germination, shoot and root lengths, fresh and dry weights, chlorophyll content, and nodules (number, weight, leghemoglobin, respiration, and nitrogenase activity) were inhibited by salinity. Ca2+ substitution for Na+, particularly at a Na/Ca ratio of 1:5, was stimulatory to almost all parameters at both stages. Statistical correlations between salinity levels and Na/Ca combinations proved one of the four levels (strong- or weak positive, strong- or weak negative) with most of the investigated parameters, depending on the parameter.

Photosynthesizing carbonate/nitrate into Chlorococcum humicola biomass for biodiesel and Bacillus coagulans-based biohydrogen production.

Biofuel can be generated by different organisms using various substrates. The green alga Chlorococcum humicola OQ934050 exhibited the capability to photosynthesize carbonate carbon, maybe via the activity of carbonic anhydrase enzymes. The optimum treatment is C:N ratio of 1:1 (0.2 mmoles sodium carbonate and 0.2 mmoles sodium nitrate) as it induced the highest dry mass (more than 0.5 mg.mL-1). At this combination, biomass were about 0.2 mg/mL-1 carbohydrates, 0.085 mg/mL-1 proteins, and 0.16 mg/mL-1 oil of this dry weight. The C/N ratios of 1:1 or 10:1 induced up to 30% of the Chlorococcum humicola dry mass as oils. Growth and dry matter content were hindered at 50:1 C/N and oil content was reduced as a result. The fatty acid profile was strongly altered by the applied C.N ratios. The defatted leftovers of the grown alga, after oil extraction, were fermented by a newly isolated heterotrophic bacterium, identified as Bacillus coagulans OQ053202, to evolve hydrogen content as gas. The highest cumulative hydrogen production and reducing sugar (70 ml H2/g biomass and 0.128 mg/ml; respectively) were found at the C/N ratio of 10:1 with the highest hydrogen evolution efficiency (HEE) of 22.8 ml H2/ mg reducing sugar. The optimum treatment applied to the Chlorococcum humicola is C:N ratio of 1:1 for the highest dry mass, up to 30% dry mass as oils. Some fatty acids were induced while others disappeared, depending on the C/N ratios. The highest cumulative hydrogen production and reducing sugar were found at the C/N ratio of 10:1.

Tuning photosynthetic oxygen for hydrogen evolution in synergistically integrated, sulfur deprived consortia of Coccomyxa chodatii and Rhodobium gokarnense at dim and high light.

In this work, tuning oxygen tension was targeted to improve hydrogen evolution. To achieve such target, various consortia of the chlorophyte Coccomyxa chodatii with a newly isolated photosynthetic purple non-sulfur bacterium (PNSB) strain Rhodobium gokarnense were set up, sulfur replete/deprived, malate/acetate fed, bicarbonate/sulfur added at dim/high light. C. chodatii and R. gokarnense are newly introduced to biohydrogen studies for the first time. Dim light was applied to avoid the inhibitory drawbacks of photosynthetic oxygen evolution, values of hydrogen are comparable with high light or even more and thus economically feasible to eliminate the costs of artificial illumination. Particularly, the consortium of 2n- (n = 1.9 × 105 cell/ml, sulfur deprived) demonstrated its perfection for the target, i.e., the highest possible cumulative hydrogen. This consortium exhibited negative photosynthesis, i.e., oxygen uptake in the light. Most hydrogen in consortia is from bacterial origin, although algae evolved much more hydrogen than bacteria on per cell basis, but for only one day (the second 24 h), as kinetics revealed. The higher hydrogen in unibacterial culture or consortia results from higher bacterial cell density (20 times). Consortia evolved more hydrogen than their respective separate cultures, further enhanced when bicarbonate and sulfur were supplemented at higher light. The share of algae relatively increased as bicarbonate or sulfur were added at higher light intensity, i.e., PSII activity partially recovered, resulting in a transient autotrophic hydrogen evolution. The addition of acetic acid in mixture with malic acid significantly enhanced the cumulative hydrogen levels, mostly decreased cellular ascorbic acid indicating less oxidative stress and relief of PSII, relative to malic acid alone. Starch, however, decreased, indicating the specificity of acetic acid. Exudates (reducing sugars, amino acids, and soluble proteins) were detected, indicating mutual utilization. Yet, hydrogen evolution is limited; tuning PSII activity remains a target for sustainable hydrogen production.

Production and optimization of bioplastic (Polyhydroxybutyrate) from Bacillus cereus strain SH-02 using response surface methodology.

BACKGROUND: Polyhydroxybutyrate (PHB) is a biopolymer formed by some microbes in response to excess carbon sources or essential nutrient depletion. PHBs are entirely biodegradable into CO2 and H2O under aerobic and anaerobic conditions. It has several applications in various fields such as medicine, pharmacy, agriculture, and food packaging due to its biocompatibility and nontoxicity nature. RESULT: In the present study, PHB-producing bacterium was isolated from the Dirout channel at Assiut Governorate. This isolate was characterized phenotypically and genetically as Bacillus cereus SH-02 (OM992297). According to one-way ANOVA test, the maximum PHB content was observed after 72 h of incubation at 35 °C using glucose and peptone as carbon and nitrogen source. Response surface methodology (RSM) was used to study the interactive effects of glucose concentration, peptone concentration, and pH on PHB production. This result proved that all variables have a significant effect on PHB production either independently or in the interaction with each other. The optimized medium conditions with the constraint to maximize PHB content and concentration were 22.315 g/L glucose, and 15.625 g/L peptone at pH 7.048. The maximum PHB content and concentration were 3100.799 mg/L and 28.799% which was close to the actual value (3051 mg/l and 28.7%). The polymer was identified as PHB using FTIR, NMR, and mass spectrometry. FT-IR analysis showed a strong band at 1724 cm- 1 which attributed to the ester group's carbonyl while NMR analysis has different peaks at 169.15, 67.6, 40.77, and 19.75 ppm that were corresponding to carbonyl, methine, methylene, and methyl resonance. Mass spectroscopy exhibited molecular weight for methyl 3- hydroxybutyric acid. CONCLUSION: PHB-producing strain was identified as Bacillus cereus SH-02 (OM992297). Under optimum conditions from RSM analysis, the maximum PHB content and concentration of this strain can reach (3100.799 mg/L and 28.799%); respectively. FTIR, NMR, and Mass spectrometry were used to confirm the polymer as PHB. Our results demonstrated that optimization using RSM is one of the strategies used for reducing the production cost. RSM can determine the optimal factors to produce the polymer in a better way and in a larger quantity without consuming time.

Bioenergetics of lactate vs. acetate outside TCA enhanced the hydrogen evolution levels in two newly isolated strains of the photosynthetic bacterium Rhodopseudomonas.

Two local hydrogen-evolving strains of purple nonsulfur bacteria have been isolated, characterized, and identified as Rhodopseudomonas sp. TUT (strains Rh1 and Rh2). Lactate followed by succinate and malate supported the highest amounts of H2 production, growth (O.D.660nm, proteins and bacteriochlorphyll contents), nitrogenase activity, and uptake hydrogenase; the least of which was acetate. Alginate-immobilized cells evolved higher hydrogen amounts than free cell counterparts. Rh1 was more productive than Rh2 at all circumstances. Lactate-dependent hydrogen evolution was more than twice that of acetate, due to ATP productivity (2/-1, respectively), which is limiting to the nitrogenase activity. The preference of lactate over other acids indicates the feasibility of using these two strains in hydrogen production from dairy wastewater.

Biofabrication strategy of silver-nanodrug conjugated polyhydroxybutyrate degrading probiotic and its application as a novel wound dressing.

Wound infections with rising incidences of multi-drug resistant bacteria are among the public health problems worldwide. The current study describes wound dressing materials made from biodegradable polyhydroxybutyrate (PHB) combined with AgNPs and gelatin (AgNPs/Gelatin/PHB). Microbial PHB was mixed with gelatin (1:2) to form a polymer matrix which was loaded with different concentrations of AgNPs (8.3-133 µg/mL). The statistical results of AgNPs synthesizing based on Box-Behnken design revealed that 1.247 mM silver nitrate and 24.054 % of Corchorus olitorius leaf extract concentration at pH (8.07) were the optimum values for the biosynthesis. UV-Vis spectroscopy, FTIR study and XRD reflects that nanoparticles are formed. The UV-Vis spectroscopy of Gelatin/PHB/AgNPs exhibited two specific bands at 298 nm and 371 nm, which confirm the formation of the conjugate. AgNPs had MICs and MBCs of (24.9, 24.9, and 12.45 µg/mL) and (33.25, 33.25, and 16.6 µg/mL) against (Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus). The MIC and MBC of AgNPs/Gelatin/PHB against the same tested bacteria were 31.1 µg and 41.5 µg, respectively. AgNPs/Gelatin/PHB exhibit excellent antimicrobial efficacy against bacteria. Sterilized gauze loaded with 31.1 µg of AgNPs/Gelatin/PHB acted as an effective wound dressing. Thus, the study highlights the importance of wound dressings developed from degradable AgNPs/Gelatin/PHB in enhancing antimicrobial efficiency and facilitating a better wound healing process.

Physical Characteristics, Mineral Content, and Antioxidant and Antibacterial Activities of Punica granatum or Citrus sinensis Peel Extracts and Their Applications to Improve Cake Quality.

One-third of all food produced for human use is discarded as waste, resulting in environmental pollution and impaired food security. Fruit peels have bioactive compounds that may be used as antimicrobials and antioxidants, and the use of fruit peels is considered an alternative way to reduce environmental problems and agro-industrial waste. The aim of this study was to evaluate the phytochemical, mineral, extraction yield, total phenolic, total flavonoids, antioxidant, and antibacterial activity of several peel fruits, including Citrus sinensis (orange) and Punica granatum (pomegranate). The results revealed that pomegranate peel powder contains the highest amounts of ash, fiber, total carbohydrates, Ca, Fe, Mg, and Cu, while orange peel contains the highest amounts of moisture, protein, crude fat, P, and K. Furthermore, the aqueous and methanolic pomegranate peel extracts yielded higher total phenolic and total flavonoids than the orange peel extract. The identification and quantification of polyphenol compounds belonging to different classes, such as tannins, phenolic acids, and flavonoids in pomegranate peel and flavonoid compounds in orange peel were performed using UPLC-MS/MS. In addition, GC-MS analysis of orange peel essential oil discovered that the predominant compound is D-Limonene (95.7%). The aqueous and methanolic extracts of pomegranate peel were proven to be efficient against both gram-positive and gram-negative bacteria linked to human infections. Sponge cake substituting wheat flour with 3% pomegranate peel and 10% orange peel powder had the highest total phenolic, flavonoid compounds, and antioxidant activity as compared to the control cake. Our results concluded that pomegranate and orange peel flour can be used in cake preparation and natural food preservers.

Bioplastic Production by Bacillus wiedmannii AS-02 OK576278 Using Different Agricultural Wastes.

Polyhydroxybutyrates (PHBs) are macromolecules synthesized by bacteria. Because of their fast degradability under natural environmental conditions, PHBs were selected as alternatives for the production of biodegradable plastics. Sixteen PHB-accumulating strains were selected and compared for their ability to accumulate PHB granules inside their cells. Isolate AS-02 was isolated from cattle manure and identified as Bacillus wiedmannii AS-02 OK576278 by means of 16S rRNA analysis. It was found to be the best producer. The optimum pH, temperature, and incubation period for the best PHB production by the isolate were 7, 35 °C, and 72 h respectively. PHB production was the best with peptone and glucose as nitrogen and carbon sources at a C/N ratio of (2:1). The strain was able to accumulate 423, 390, 249, 158, and 144 mg/L PHB when pretreated orange, mango, banana, onion peels, and rice straw were used as carbon sources, respectively. The extracted polymer was characterized by Fourier transform infrared (FTIR), nuclear magnetic resonance (NMR), and GC-MS spectroscopy, which confirmed the structure of the polymer as PHB. The isolate B. wiedmannii AS-02 OK576278 can be considered an excellent candidate for industrial production of PHB from agricultural wastes.