An alkaliphilic-thermotolerant Bacillus cereus N1 isolated from Bani Salama Lake, Wadi El-Natron, Egypt, was proved to produce mannanase enzyme. Optimization of the fermentation medium components using Plackett–Burman design was applied. Glucose and inoculum size were found to be the most significant factors enhancing the production of the enzyme. On applying optimized medium in the fermentation process, an enzyme productivity of 42.2 UmL−1 was achieved with 6.4 fold increase compared to the basal one. Mannanase was also extracted and purified using chromatography such as ion-exchange chromatographic and gel filtration methods. It was indicated that, the mannanase activity extracted and purified from the isolate B. cereus N1 was reduced to 321.6 U (about 36% of the whole mannanase in the culture filtrate) in comparison with the initial mannanase activity (900 U) and the total protein content reduced to 52 mg (the initial total protein content was 220 mg). However, the specific activity for the mannanase from B. cereus N1 at the end of the purification steps was found to be about 628 Umg−1 compared to 4.2 Umg−1 at the initial culture filtrate. It was also indicated that the mannanase enzyme was purified almost 149-fold.

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Twenty fluorescent Pseudomonas isolates were tested for their ability to produce siderophores on chrome azurol S (CAS) agar plates and their antagonistic activity against six plant pathogenic fungal isolates was assessed. Scaling-up production of siderophores from the promising isolates, P. aeruginosa F2 and P. fluorescens JY3 was performed using batch and exponential fed-batch fermentation. Finally, culture broth of the investigated bacterial isolates was used for the preparation of two economical bioformulations for controlling Fusarium oxysporum and Rhizoctonia solani. The results showed that both isolates yielded high siderophore production and they were more effective in inhibiting the mycelial growth of the tested fungi compared to the other bacterial isolates. Exponential fed-batch fermentation gave higher siderophore concentrations (estimated in 10 µL), which reached 67.05% at 46 h and 45.59% at 48 h for isolates F2 and JY3, respectively, than batch fermentation. Formulated P. aeruginosa F2 and P. fluorescens JY3 decreased the damping-off percentage caused by F. oxysporum with the same percentage (80%), while, the reduction in damping-off percentage caused by R. solani reached 87.49% and 62.5% for F2 and JY3, respectively. Furthermore, both formulations increased the fresh and dry weight of shoots and roots of wheat plants. In conclusion, bio-friendly formulations of siderophore-producing fluorescent Pseudomonas isolates can be used as biocontrol agents for controlling some plant fungal diseases.

In this study, a trial was made to control the infection of apple plants by Rhizoctonia solani AG-5. Five soil amendments: compost, cow manure, chicken manure, Brassica juncea seed meal and urea, were applied to Columbia View Experimental soil planted with six-week-old apple transplants. Soil was applied in two forms, pasteurized and nonpasteurized. The results showed that the amendments enhanced the increase in microbial count. The effect was more pronounced in fungi and pseudomonads. The increase in the fungal count was particularly clear in the case of B. juncea seed meal and urea, leading to 40-and 20-fold increase, respectively. Regarding the pseudomonads count, the same two amendments, B. juncea seed meal and urea, resulted in 76.5-and 54-fold increase, respectively. It was also observed that apple seedlings planted in pasteurized soil started to wilt and die faster than those planted in non-pasteurized soil, regardless of the type of amendment supplied. Brassica juncea seed meal showed the most pronounced positive effect when added to non-pasteurized soil, which could completely prevent the infection of apple seedlings. In tests to detect the DNA of R. solani AG-5 in the roots of apple replants by real-time polymerase chain reaction, it was observed that only B. juncea seed meal and urea could significantly reduce the pathogen DNA level in the pasteurized soil and decreased it to a zero level in case of non-pasteurized soil.

A Gram-positive, rod-shaped, spore-forming haloalkaliphilic bacterium designated as NA7 was isolated from the surface of a Helianthemum nummularium root sample obtained from Wadi Natrun in Egypt. Sequence analysis of the 16S rRNA gene revealed a Bacillus haloalkaliphilius strain as the closest match with 99% identity. In a shake flask culture containing 10% NaCl, adjusted to pH 10 and incubated at 37°C, the isolated strain produced thermostable extracellular alkaline protease with relatively stable maximum activity records (0.610-0.625 TU) within a relatively long stationary phase that exceeded 60 h. A 2-level fractional factorial design (Plackett-Burman) was then applied to screen for nutritional and cultivation factors regulating protease production by the isolate and to appraise their effects. Calculated statistical parameters revealed that NaCl and MgSO 4 are the most significant independent variables affecting alkaline protease production by NA7 and suggested a near-optimum culture condition. Verification of this predicted condition resulted in an alkaline protease specific activity record of 509 TU/mg protein with a 1.27 fold increase when compared to the basal medium culture.

A total of eight cultivable purified Egyptian yeast phenotypes were isolated from different sources, including fruits, juices and paste, and were compared to a baker's yeast factory reference strain. Genotypic characterization of the most promising new isolate (RO1) confirmed its identification as a Saccharomyces cerevisiae strain. In a shake-flask experiment, the PlackettÀBurman multi-factorial design was applied to identify factors that considerably affect the RO1 growth rate. Together with the components of the factory molasses-based medium, six other culture factors, hypothesized to affect yeast biomass production, were examined as independent variables. The calculated main effect results and P-values suggested that by increasing the level of molasses, diammonium phosphate and inoculum size, compared to the factory settings, and by supplementing the medium with yeast extract, calcium pantothenate (vitamin B5) and trace elements, the RO1 biomass production was improved. Application of the predicted near-optimum fermentation conditions with scaling up the culture medium to 22 L in a 40 L airlift bioreactor resulted in 93 g L ¡1 biomass production, which represented approximately a 1.5-fold increase, when compared to the reference culture condition. Moreover, the dough raising test indicated that the newly isolated yeast strain RO1 caused a 1.75-fold increase in the fermentative power, when compared to the factory reference strain.

Background Cholesterol oxidases (CHOs) have attracted enormous attention because of their wide biotechnological potential. The present study explores the production of CHOs by Streptomyces sp. AN. Evaluation of culture conditions affecting enzyme production, medium optimization and released metabolite characteristics were also investigated. Results The current work reports the isolation of 37 colonies (bacteria/actinobacteria) with different morphotypes from different soil/water samples. The isolate-coded AN was selected for its high potency for CHO production. Morphological characteristics and the obtained partial sequence of 16srRNA of AN showed 99.38% identity to Streptomyces sp. strain P12–37. Factors affecting CHO production were evaluated using Plackett-Burman (PB) and Box-Behnken (BB) statistical designs to find out the optimum level of the most effective variables, namely, pH, starch, NH4NO3 and FeSO4.7H2O with a predicted activity of 6.56 U/mL. According to this optimization, the following medium composition was considered to be optimum (g/L): cholesterol 1, starch 6, MgSO4.7H2O 0.1, CaCl2 0.01, FeSO4.7H2O 0.1, NH4NO3 23.97, yeast extract (YE) 0.2, K2HPO4 0.01, KH2PO4 0.1, NaCl 0.01, Tween 20 0.01, pH 6.36 and incubation temperature (30 °C) for 9 days. Spectophotometric analysis for released metabolites against cholesterol (standard) via Fourier-transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC) was carried out. FTIR spectrum showed the appearance of new absorption peaks at 1644 and 1725cm−1; this confirmed the presence of the Keto group (C=O) stretch bond. Besides, fermentation caused changes in thermal properties such as melting temperature peak (99.26; 148.77 °C), heat flow (− 8; − 3.6 Mw/mg), capacity (− 924.69; − 209.77 mJ) and heat enthalpy (− 385.29; 69.83 J/g) by comparison to the standard cholesterol as recognized through DSC thermogram. These changes are attributed to the action of the CHO enzyme and the release of keto derivatives of cholesterol with different properties. Conclusion Streptomyces sp. AN was endowed with the capability to produce CHO. Enzyme maximization was followed using a statistical experimental approach, leading to a 2.6-fold increase in the overall activity compared to the basal condition. CHO catalyzed the oxidation of cholesterol; this was verified by the appearance of a new keto group (C=O) peak at 1644 and 1725 cm−1 observed by FTIR spectroscopic analysis. Also, DSC thermogram demonstrates the alteration of cholesterol triggered by CHO.

Secretion of siderophores by Pseudomonas aeruginosa F2 and P. fluorescens JY3 was evaluated on chrome azurol S (CAS) agar plates and their inhibitory effect was inspected against Fusarium oxysporum and Rhizoctonia solani. Production of siderophores as biocontrol agents from F2 and JY3 was accomplished in two optimized media. Afterward, cell-free supernatants of the bacterial cultures containing siderophores were used for the preparation of two bio-friendly formulations for the management of F. oxysporum and R. solani under greenhouse conditions. The investigated bacterial isolates, F2 and JY3, showed antagonistic activity in vitro against F. oxysporum and R. solani and produced siderophores in optimized media with high efficiency. Colonies of both bacterial isolates were grown exponentially with a constant specific growth rate of 0.07 h−1 and 0.27 h−1, correspondingly. Siderophores estimated in 10 µL reached their highest value of 16.95% at 47 h and 19.5% at 48 h for isolate F2 and JY3, respectively. Formulations of siderophore-generating F2 and JY3 reduced damping-off caused by F. oxysporum by 40% and 80%, while the reduction percentage of damping-off caused by R. solani reached 87.5% and 62.5%, correspondingly. Moreover, both formulations encouraged the growing of wheat plants where the fresh and dry weight of shoots and roots were increased compared to the treatment with each fungus. In conclusion, bio-friendly formulations resulting from this investigation can play an active role in managing soil-borne diseases.