The Plant Growth-Promoting Bacteria Strain Bacillus mojavensis I4 Enhanced Salt Stress Tolerance in Durum Wheat.

Plant growth and production are adversely affected by soil salinity. A plant growth-promoting bacteria (PGPB) designated as the "I4 strain" of Bacillus mojavensis was isolated from Tunisian soil (Sfax, Tunisia) and showed the ability to be grown in the presence of NaCl concentrations ranging from 0 to 10% in Luria Bertani (LB) medium. The PGPB-mediated salt tolerance in durum wheat was evaluated. The physiological parameters such as growth, shoot and root length, dry and fresh weight were higher in I4-inoculated wheat plants in comparison with non-treated plants under salt stress. Results showed that this strain promoted wheat growth and preserved the membrane damage by notably lowering the electrolytes leakage and malondialdehyde content in contrast to non-inoculated plants. Moreover, leaf chlorophyll content, biochemical parameters and antioxidant enzyme activities measurement showed a better salt and heavy metal stress adaptation of the I4-inoculated plants. Due to these outcomes, it could be suggested that the inoculation of the PGPB I4 strain enhanced the wheat plant's growth, especially under salt stress conditions. This study confirms the ameliorative role played by PGPB in tolerating salt stress in wheat and their potential use as biofertilizers to enhance its growth in saline soil and help in promoting this plant's culture to provide food security under these perturbed global circumstances.

Antimicrobial and antioxidant activities of Bacillus mojavensis I4 lipopeptides and their potential application against the potato dry rot causative Fusarium solani.

Lipopeptides are diverse metabolites produced by various bacterial and fungal genera. They are known for their antimicrobial and surfactant activities with diverse environmental, pharmaceutical, and also agronomic applications as biocontrol agents. In this study, a PCR was used to confirm the presence of NRPS genes in Bacillus mojavensis I4. This bacterial strain could produce diverse lipopeptides which belong to the fengycin, and surfactin families. The antioxidant activity of I4 biosurfactants was determined through four different in vitro assays. Furthermore, antimicrobial activity assays indicated that I4 lipopeptides exhibited marked inhibitory activity against several bacterial and fungal strains. Further treatment of potato dry rot causative pathogen Fusarium solani with I4 lipopeptides demonstrated a remarkable reduction in the fungal penetration by almost 80% after 15 days of incubation. The findings suggest that I4 lipopeptide is a potential biocontrol agent during potato tuber storage.

Isolation of bacterial strains from compost teas and screening of their PGPR properties on potato plants.

The beneficial effect of compost and compost tea on plant growth and protection is mainly associated with the microbial diversity and the presence of bacteria with plant growth-promoting effect. PGPR are considered as eco-friendly bio-fertilizers that may reduce the use of chemical pesticides and fertilizers. Three composts (AT, A10, and A30) were previously prepared from industrial wastes (olive mill wastewater, olive pomace, coffee ground, and phosphogypsum). In the present study, we isolated three bacterial strains from the compost teas. The phylogenetic identification of these bacterial strains (B.AT, B.A10, and B.A30) showed that they correspond to Serratia liquefaciens (B.AT and B.A10) and Achromobacter spanius (B.A30) species. A further characterization of the PGPR traits of these bacteria showed that they produce siderophore, exopolysaccharides, and IAA. Their effect on potato plant growth, yields, and tuber quality was performed under field culture conditions. Results showed that these strains can be characterized as PGPR, the best effect on potato plant growth was observed with Serratia liquefaciens (B.AT), the best yield and tuber quality was observed with Serratia liquefaciens (B.A10) while bacterial treatment with Achromobacter spanius (B.A30) is a Cd-tolerant PGPR.

Characterization and production optimization of biosurfactants by Bacillus mojavensis I4 with biotechnological potential for microbial enhanced oil recovery.

Response surface methodology was applied to optimize the production of biosurfactants from Bacillus mojavensis I4 using Box-Behnken design with four variables. The optimal variable combination was 3% of glucose as carbon source, 0.6% of glutamic acid as nitrogen source, temperature of 35 °C and 10 g/l of NaCl which yielded to an optimal production of 4.12 g/l. Compositional analysis and FTIR spectrum revealed that the extracted biosurfactants was a lipopeptides. The biosurfactants achieved a critical micelle concentration value of 100 mg/l. Moreover, the extracted biosurfactants were effective at recovering up to 89.2% of motor oil from sand beach and achieved a dispersion rate of 78% of the initial diameter of the oil. These findings suggested the potential use of I4 biosurfactants in the oil industry.

Investigation of the response to salinity of transgenic potato plants overexpressing the transcription factor StERF94.

Salinity is one of the most important constraints threatening the cultivation of potato plants (Solanum tuberosum L.). It affects plant growth and leads to significant yield loss. Consequently, it is important to improve the tolerance of potato plants to salinity. In this context, we investigated the involvement of a potato ethylene responsive factor (StERF94) in plant response to salinity, since our previous genome-wide analysis showed that it may be related to biotic and abiotic stress response. ERF proteins belong to a large family of transcription factors that participate in plant response to abiotic stresses. We have previously identified the StERF94 gene which shows increased expression in potato plants submitted to salt treatment. In this study, transgenic potato plants overexpressing StERF94 were produced and submitted to salt treatment (100 mM NaCl) in vitro and under greenhouse culture conditions. StERF94 transgenic lines showed lower decrease of stem elongation under salt treatment in comparison to non-transgenic wild-type plants. Moreover, these plants showed a low level of H2O2 and Malondialdehyde content, and an increase in catalase and GPX (Gluthation peroxidase) activities compared to non-transgenic plants. In a second step, enhanced expression of some target genes for example CuZn-SOD, DHN25 (Dehydrin) and ERD (Early Responsive to Dehydration) was noted in the StERF94 transgenic plants, submitted to salt treatment. The StERF94 factor was also involved in the activation of osmoprotectant synthesis. Taken together, all these data suggest that overexpression of the StERF94 transcription factor increases the tolerance of potato plants to salinity by improving plant growth, osmoprotectant synthesis and antioxidant activityleading to low oxidative stress damage.

Improved Biosurfactant Production by Bacillus subtilis SPB1 Mutant Obtained by Random Mutagenesis and Its Application in Enhanced Oil Recovery in a Sand System.

Biosurfactants or microbial surfactants are surface-active biomolecules that are produced by a variety of microorganisms. Biodegradability and low toxicity have led to the intensification of scientific studies on a wide range of industrial applications for biosurfactants in the field of environmental bioremediation as well as the petroleum industry and enhanced oil recovery. However, the major issues in biosurfactant production are high production cost and low yield. Improving the bioindustrial production processes relies on many strategies, such as the use of cheap raw materials, the optimization of medium-culture conditions, and selecting hyperproducing strains. The present work aims to obtain a mutant with higher biosurfactant production through applying mutagenesis on Bacillus subtilis SPB1 using a combination of UV irradiation and nitrous acid treatment. Following mutagenesis and screening on blood agar and subsequent formation of halos, the mutated strains were examined for emulsifying activity of their culture broth. A mutant designated B. subtilis M2 was selected as it produced biosurfactant at twice higher concentration than the parent strain. The potential of this biosurfactant for industrial uses was shown by studying its stability to environmental stresses such as pH and temperature and its applicability in the oil recovery process. It was practically stable at high temperature and at a wide range of pH, and it recovered above 90% of motor oil adsorbed to a sand sample.

Potential application of Bacillus subtilis SPB1 biosurfactants in laundry detergent formulations: Compatibility study with detergent ingredients and washing performance.

Surfactants play a very important role in laundry and household cleaning products ingredients. In this research, the application of lipopeptide biosurfactants, produced by Bacillus subtilis SPB1, in the formulation of a washing powder was investigated. The SPB1 biosurfactant was mixed with sodium tripolyphosphate as a builder and sodium sulfate as filler. The efficiency of the formulated detergent composition with different washing conditions to remove a stain from cotton fabric was examined. The results showed that the formulated detergent was effective in oil removal, with optimal washing conditions of pH, temperature, striate and time of washing system of 7, 65°C, 1000 RPM and 60 min, respectively. A comparative study of different detergent compositions (biosurfactant-based detergent, combined biosurfactant-commercial detergent, and a commercial detergent) for the removal of oil and tea stains, proved that the bio-scouring was more effective (>75%) in terms of the stain removal than the commercial powders (<60%). Moreover, the results demonstrated that the biosurfactant acts additively with a commercial detergent and enhances their performance from 33 to 45% in removing oil stain and from 57 to 64% in removing tea stain. As a conclusion, in addition to the low toxicity and the high biodegradability of the microbial biosurfactants, the results of this study have shown that the future use of this lipopeptide biosurfactant as laundry detergent additive is highly promising.

Optimization of polysaccharides extraction from watermelon rinds: Structure, functional and biological activities.

In the present work, optimization of hot water extraction, structural characteristics, functional properties, and biological activities of polysaccharides extracted from watermelon rinds (WMRP) were investigated. The physicochemical characteristics and the monosaccharide composition of these polysaccharides were then determined using chemical composition analysis, Fourier transform infrared (FT-IR) spectroscopy, scanning electron microscopy (SEM) and gas chromatography-flame ionization detection (GC-FID). SEM images showed that extracted polysaccharides had a rough surface with many cavities. GC-FID results proved that galactose was the dominant sugar in the extracted polysaccharides, followed by arabinose, glucose, galacturonic acid, rhamnose, mannose, xylose and traces of glucuronic acid. The findings revealed that WMRP displayed excellent antihypertensive and antioxidant activities. Those polysaccharides had also a protection effect against hydroxyl radical-induced DNA damage. Functional properties of extracted polysaccharides were also evaluated. WMRP showed good interfacial dose-dependent proprieties. Overall, the results suggested that WMRP presents a promising natural source of antioxidants and antihypertensive agents.

Anionic lipopeptides from Bacillus mojavensis I4 as effective antihypertensive agents: Production, characterization, and identification.

A new isolated Bacillus mojavensis strain I4 was found as producer of biosurfactants by different screening methods, such as parafilm M test, hemolytic activity, oil displacement test, emulsification index, surface tension, and lipase production assay. Enhanced biosurfactants production was obtained using glucose and glutamic acid as carbon and nitrogen sources, respectively. The optimal production of the biosurfactants was obtained by using a C/N ratio of 17, pH of 7.0, and temperature of 37°C. The surface tension was reduced to 29 mN/m and the emulsification index E24 of 62% was achieved after 72 h of culture. The purified biosurfactants showed stability with regard to surface tension reduction and emulsification in a wide range of temperatures (4-120°C), pH (4-10), and salinity (2-12% of NaCl). The thin-layer chromatography showed that the produced biosurfactants were lipopeptides. The biosurfactants were characterized as a group of anionic lipopeptides with zeta potential measurement. Chromatographic characterization using HPLC revealed that I4 lipopeptides contained numerous isoforms and surfactin was the major component. Moreover, the I4 lipopeptides showed interesting angiotensin-converting enzyme-inhibitory activity.

Screening and Molecular Identification of New Microbial Strains for Production of Enzymes of Biotechnological Interest

ABSTRACT: This research focused on isolation, identification and characterization of new strains of fungi and bacteria, which were able to produce extracellular xylanase, mannanase, pectinase and α-amylase. Fungi isolates were identified on the basis of analyses of 18S gene sequencing and internal transcribed spacer region. The closest phylogenetic neighbors according to 18S gene sequence and ITS region data for the two isolates M1 and SE were Aspergillus fumigatus and Aspergillus sydowii, respectively. I4 was identified as Bacillus mojavensis on the basis of the 16S rRNA gene sequencing and biochemical properties. The enzyme production was evaluated by cultivating the isolated microorganisms in liquid-state bioprocess using wheat bran as carbon source. Two fungi (M1, and SE) and one bacterium (I4) strains were found to be xylanase producer, and several were proven to be outstanding producers of microbial xylanase. The strains producing xylanase secreted variable amounts of starch-debranching enzymes and produced low level β-mannan-degrading enzyme systems. The bacterium strain was found to be capable of producing pectinolytic enzymes on wheat bran at high level. Some of the strains have good potential for use as sources of important industrial enzymes.

Assessment of pectinase production by Bacillus mojavensis I4 using an economical substrate and its potential application in oil sesame extraction.

Carrot (Daucus carota) peels, local agricultural waste product, is rich in lignocellulolytic material, including pectin which can act as an inducer of pectinase production. Pectinolytic enzymes production by Bacillus mojavensis I4 was studied in liquid state fermentation using carrot peel as a substrate. Medium composition and culture conditions for the pectinase production by I4 were optimized using two statistical methods: Taguchi design was applied to find the key ingredients and conditions for the best yield of enzyme production and The Box-Behnken design was used to optimize the value of the four significant variables: carrot peels powder, NH4Cl, inoculum size and incubation time. The optimal conditions for higher production of pectinase were carrot peels powder 6.5 %, NH4Cl 0.3 %, inoculum level 3 % and cultivation time 32 h. Under these conditions, the pectinase experimental yield (64.8 U/ml) closely matched the yield predicted by the statistical model (63.55 U/ml) with R (2) = 0.963. The best pectinase activity was observed at the temperature of 60 °C and at pH 8.0. The enzyme retained more than 90 % of its activity after 24 h at pH ranging from 6.0 to 10.0. The enzyme preserved more than 85 % of its initial activity after 60 min of pre-incubation at 30-40 °C and more than 67 % at 50 °C. The extracellular juice of I4 was applied in the process of sesame seeds oil extraction. An improvement of 3 % on the oil yield was obtained. The findings demonstrated that the B. mojavensis I4 has a promising potential for future use in a wide range of industrial and biotechnological applications.

Water-soluble polysaccharides from agro-industrial by-products: functional and biological properties.

Water-soluble polysaccharides were isolated from almond (AWSP) and pistachio (PWSP) juice processing by-products. Their chemical and physical characteristics were determined using NMR and Infrared spectroscopic analysis. The complexities of the spectra reflected the heterogeneity of these polysaccharides. The ACE inhibitory activities (IC50 AWSP=2.81mgmL(-1) and IC50 PWSP=2.59mgmL(-1)) and antioxidant properties of AWSP and PWSP were investigated based on the DPPH radical-scavenging capacity assay (IC50 AWSP=2.87mgmL(-1) and IC50 PWSP=1.61mgmL(-1)). Reducing power, ß-carotene bleaching inhibition (IC50AWSP=4.46mgmL(-1) and IC50 PWSP=3.39mgmL(-1)), and ferrous chelating assays (IC50 AWSP=0.22mgmL(-1) and IC50 PWSP=0.19mgmL(-1)) were also performed. The findings revealed that water-soluble polysaccharides exhibited antioxidant and antihypertensive activities. AWSP and PWSP showed excellent interfacial concentration-dependent properties. Overall, the results suggested that both AWSP and PWSP are promising sources of natural antioxidants and ACE inhibitory agents and could, therefore, be used as alternative additives in food, pharmaceutical and cosmetic preparations.