Date Powder as Alternative Carbone Source in Optimizing the Cultivation of (Bacillus megaterium) in Submerged State Fermentation

Problems statement: A study had shown a strain of Bacillus megaterium was isolated from the soil. Questions were whether date powder could be used as alternatives of carbon sources for Bacillus megaterium submerged state cultivation? Approach: To evaluate the effects of the using date powder; The fermentation was carried out in shake flasks using a complex medium consisting of (g/L) ammonium tartrate, 2.0; magnesium sulphateheptahydrate, 4.0; dipotassium phosphate, 14.0; calcium chloride, 0.2, NaHPO4, 4.0, yeast extract, 3.0, trace elements, 2.0ml and glucose 6.0(4.0, yeast extract, 3.0, trace elements, 2.0ml and glucose 6.0) were replaced by dates powder at three rates; 5,10 and15 g/L Results show that the rate of production of biomass during submerged fermentation by Bacillus megaterium at 30±1°C for 24 hours of incubation using date powder was maximizes between 18 and 24 hr of fermentation. The experimental runs and results for the Box– Behnken design. The 15 runs in a single block were used to study the effects using date powder in different rates on one response. Biomass concentration ranged from 6.5cfu /l to 12.9cfu /.comparing with 14.8 cfu/ g for the commercial growth media. The ANOVA tables give the statistical significance of the effects for biomass. These effects were dates powder rates, the optimized growth medium with pH 7, inoculums’ size 106, temp 24oC and moisture dates powder 15 g/L. The biomass concentrations obtained under the optimal conditions 12.9cfu/l. Key words: Date Powders, Fermentation, Bacillus megaterium, biomass, Submerged State

Estimation of the Efficiency of Methods for Electroplating Wastewater Purification from Ammonium-Tartrate Copper (II) Complexes

Wastewater purification from heavy metal compounds is a complex and urgent task. One of the main sources of pollution of the environment with ions of heavy metals is the wastewater, i.e., washing, of the electro-plating processes. Complex electrolytes based on copper compounds are stable in a wide pH range and, when released into water, cannot be removed by the traditional methods, such as neutralization and precipitation. The study estimated the efficiency of various methods of physicochemical water purification for removing complex ammonia-tartrate copper (II) complexes from water. Findings of research show that titanyl sulfate is most effective in water purification using coagulants. The efficiency of purification with the use of titanium compounds reaches 85 %, which is on average 30--40 % higher than when using traditional coagulants based on aluminum or iron compounds. Electrocoagulation processes make it possible to effectively remove complex copper compounds from water due to a combination of the processes of organic component oxidation and coagulation with iron salts. It was found that advanced oxidation processes, which are based on the reaction with a hydroxyl radical, using hydrogen peroxide, i.e., Fenton processes, make it possible to purify wastewater from copper compounds by 99.9 %. Despite the high efficiency, it is advisable to use adsorption processes only at the stage of additional water purification from previously coagulated and oxidized pollutants

Corn and Soybean Host Root Endophytic Fungi with Toxicity Toward the Soybean Cyst Nematode

Although fungal endophytes are commonly investigated for their ability to deter microbial plant pathogens, few studies have examined the activity of fungal root endophytes against nematodes. The soybean cyst nematode (SCN; Heterodera glycines), the most severe yield-limiting pathogen of soybean (Glycine max), is commonly managed through rotation of soybean with corn (Zea mays), a nonhost of the SCN. A total of 626 fungal endophytes were isolated from surface-sterilized corn and soybean roots from experimental plots in which soybean and corn had been grown under annual rotation and under 1, 3, 5, and 35 years of continuous monoculture. Fungal isolates were grouped into 401 morphotypes, which were clustered into 108 operational taxonomic units (OTUs) based on 99% sequence similarity of the full internal transcribed spacer region. Morphotype representatives within each OTU were grown in malt extract broth and in a secondary metabolite-inducing medium buffered with ammonium tartrate, and their culture filtrates were tested for nematicidal activity against SCN juveniles. A majority of OTUs containing isolates with nematicidal culture filtrates were in the order Hypocreales, with the genus Fusarium being the most commonly isolated nematicidal genus from corn and soybean roots. Less commonly isolated taxa from soybean roots included the nematophagous fungi Hirsutella rhossiliensis, Metacordyceps chlamydosporia, and Arthrobotrys iridis. Root endophytic fungal diversity in soybean was positively correlated with SCN density, suggesting that the SCN plays a role in shaping the soybean root endophytic community.

Crystallization of the rice immune receptor RGA5A_S with the rice blast fungus effector AVR1-CO39 preparedviamixture and tandem strategies

RGA5 is a component of the Pia resistance-protein pair (RGA4/RGA5) fromOryza sativaL.japonica. It acts as an immune receptor that directly recognizes the effector AVR1-CO39 fromMagnaporthe oryzae viaa C-terminal non-LRR domain (RGA5A_S). The interaction between RGA5A_S and AVR1-CO39 relieves the repression of RGA4, leading to effector-independent cell death. To determine the structure of the complex of RGA5A_S and AVR1-CO39 and to understand the details of this interaction, the complex was prepared by fusing the proteins together, by mixing themin vitroor by co-expressing them in one host cell. Samples purifiedviathe first two strategies were crystallized under two different conditions. A mixture of AVR1-CO39 and RGA5A_S (complex I) crystallized in 1.1 Mammonium tartrate dibasic, 0.1 Msodium acetate–HCl pH 4.6, while crystals of the fusion complex RGA5A_S-TEV-AVR1-CO39 (complex II) were grown in 2 MNaCl. The crystal of complex I belonged to space groupP3121, with unit-cell parametersa = b= 66.2,c= 108.8 Å, α = β = 90, γ = 120°. The crystals diffracted to a Bragg spacing of 2.4 Å, and one molecule each of RGA5A_S and AVR1-CO39 were present in the asymmetric unit of the initial model. The crystal of complex II belonged to space groupI4, with unit-cell parametersa=b= 137.4,c= 66.2 Å, α = β = γ = 90°. The crystals diffracted to a Bragg spacing of 2.72 Å, and there were two molecules of RGA5A_S and two molecules of AVR1-CO39 in the asymmetric unit of the initial model. Further structural characterization of the interaction between RGA5A_S and AVR1-CO39 will lead to a better understanding of the mechanism underlying effector recognition by R proteins.

MEDIUM OPTIMIZATION FOR THE PRODUCTION OF BIOMASS BY Cunninghamella sp. 2A1 USING RESPONSE SURFACE METHODOLOGY

A statistical design approach has been used to optimize the production of biomass byCunninghamellasp. 2A1, evaluated based on lipidless biomass. A 23full factorial centralcomposite design (CCD) was chosen to study the combined effects of three factors; ammoniumtartrate, peptone and glucose concentrations. The p-value for each factors was < 0.05 suggestingthat these factors have significant effect on the production of lipidless biomass. The productionis represented by a linear model with p-value < 0.0001. The optimized medium consists of 3.86g/l ammonium tartrate, 55.84 g/l glucose and 7.73 g/l peptone predicted 16.83 g/l lipidlessbiomass. Results from four replications based on the optimized medium produced 18.48 g/llipidless biomass, which are in close agreementwith the predicted value. The coefficient forglucose was the highest indicating it as the most significant factoraffecting lipidless biomassproduction.

Nutrient and Physiological Requirements for Biomass Production of Pestalotiopsis sp. UMAS P14 and Pseudopestalotiopsis sp. UMAS P2005/2592

Fungi are important decomposers in our ecosystem and are useful in metabolite production, bio-degradation and bio-sorption of different substrates and wastes through their mycelial biomass. Fungal species are known to have different preferences for their growth requirements. Two fungal species Pestalotiopsis and Pseudopestalotiopsis useful in many biotechnological applications were studied. Nutrient and physiological requirements for mycelial biomass production such as carbon and nitrogen, pH, light and temperature were investigated. Both fungal isolates were grown in liquid basal medium supplemented separately with various carbon and nitrogen source and incubated under different light, pH and temperature conditions for 15 days. In general, Pestalotiopsis sp. and Pseudopestalotiopsis sp. showed significant preferences for monosaccharide and disaccharide carbon source as compared to sugar alcohols and polysaccharides, whereas ammonium tartrate was more preferred as a nitrogen source compared to ammonium nitrate, ammonium sulphate and other ammonium salts. These two fungal species were able to grow and produce good mycelial biomass (223.33 mg for Pseudopestalotiopsis and 136.67 mg for Pestalotiopsis) at temperature range of 15°C to 30°C and (290.00 mg for Pseudopestalotiopsis and 256.67 mg for Pestalotiopsis) on media pH of slightly acidic to slightly alkaline. However, they showed no significant preferences between constant light, total darkness and alternate light conditions. The results from this study will be very useful for the mycelial biomass production of Pestalotiopsis sp. and Pseudopestalotiopsis sp. for their biotechnological applications.