Optimization of extracellular catalase production from Aspergillus phoenicis K30 by a linear regression method using date flour as single carbon source and purification of the enzyme

Aspergillus phoenicis K30 is the selected mutant which produces an amount of extracellular catalase. To amplify the extracellular catalase production by the strain, a fermentation optimization was performed. To select the factors affecting the production, nine active variables (factors) consisting of 12 experiments were analyzed by Plackett-Burman design. Each variable was tested at two levels, a higher and a lower level. The studies of the effect of each variable and the establishment of a correlation between the response of enzyme activity and variables revealed that the link is a multiple linear regression form. The optimization was carried out through a simplex algorithm. The amount of extracellular catalase produced by the strain in the optimized medium was about four times higher than that obtained in non optimized medium corresponding to 3820 mg/L of extracellular proteins including 59500 U/L of extracellular catalase activity after 96 h of fermentation. The steps of purification were allowed to improve enzyme activity by 305-fold. From an analytical gel electrophoresis under native conditions, an apparent molecular mass of 158 kDa was determined suggesting that the enzyme is a homodimer. The isoelectric point of the protein was found to be 5 ± 0.1 as determined by a Pharmacia Phast-system.Keywords: Aspergillus phoenicis, extracellular catalase purification, dates flour, optimization, multiple linear regression.African Journal of Biotechnology Vol. 12(19), pp. 2646-265

Decommissioned dates: chemical composition and fermentation substrate for the production of extracellular catalase by an Aspergillus phoenicis mutant

peer reviewedThe recovery of dates downgraded as a fermentation medium for the production of extracellular catalase by Aspergillus phoenicis K30 was studied. Analysis of the chemical composition of pulp and kernel flour of dates showed that the pulp had a considerably greater carbohydrate content compared to the kernel (84 vs 2.93% respectively). However, the kernel flour was richer in nitrogen (0.68% vs 0.34), mineral elements (3.63 vs 1.28%) and in essential fatty acids C18: 2 vs C18: 3 than the pulp flour. The soluble extract of the date flour showed that sugars solubilised at 90% consisted of sucrose, fructose and glucose. Therefore, this extract, being an important source of carbon and energy, was used in the current study as a fermentation medium (after supplementation with 20% of corn steep) for the production of extracellular catalase by A. Phoenicis K30. During the course of this fermentation, the biomass was estimated at 18.6 g / L after 72 h of culture, while the maximum concentration of extracellular catalase (47.5 U / ml) was reached at 96 h of fermentation. The mycelium obtained in pellet form is suitable for industrial exploitation of this process

Optimization of thermostable proteases production under agro-wastes solid-state fermentation by a new thermophilic Mycothermus thermophilus isolated from a hydrothermal spring Hammam Debagh, Algeria

The present work investigates for the first time the presence and isolation of the thermophilic fungi from hydrothermal spring situated at the locality of Guelma, in the Northeast of Algeria. The production of the thermostable proteases and the optimization of culture conditions under agro-wastes solid-state fermentation to achieve optimal production capacity were explored. A statistical experimental approach consisting of two designs was used to determine the optimum culture conditions and to attain the greatest enzyme production. Besides, different agricultural wastes were initially evaluated as a substrate, whereby wheat bran was selected for enzyme production by the isolate under solid-state conditions. The isolate thermophilic fungi were identified as Mycothermus thermophilus by sequencing the ITS region of the rDNA (NCBI Accession No: MK770356.1). Among the various screened variables: the temperature, the inoculum size, and the moisture were proved to have the most significant effects on protease activity. Employing two-level fractional Plackett?Burman and a Box?Behnken designs statistical approach helped in identifying optimum values of screened factors and their interactions. The analysis showed up 6.17-fold improvement in the production of proteases (~1187.03 U/mL) was achieved under the optimal conditions of moisture content 47%, inoculum 5 ? 105 spores/g, and temperature at 42 C. These significant findings highlight the importance of the statistical design in isolation of Mycothermus thermophilus species from a specific location as well as identifying the optimal culture conditions for maximum yield.Scopu

In Vitro and In Vivo Characterization of Plant Growth Promoting Bacillus Strains Isolated from Extreme Environments of Eastern Algeria.

This report is to our knowledge the first to study plant growth promotion and biocontrol characteristics of Bacillus isolates from extreme environments of Eastern Algeria. Seven isolates of 14 (50 %) were screened for their ability to inhibit growth of some phytopathogenic fungi on PDA and some roots exudates. The bacteria identification based on 16S r-RNA and gyrase-A gene sequence analysis showed that 71 % of the screened isolates belonged to Bacillus amyloliquefaciens and the rest were closely related to B. atrophaeus and B. mojavensis. Most of them had high spore yields (22 x 108-27 x 108 spores/ml). They produced protease and cellulase cell wall-degrading enzymes while the chitinase activity was only observed in the B. atrophaeus (6SEL). A wide variety of lipopeptides homologous was detected by liquid chromatography-electrospray ionization-mass spectrometry analysis. Interestingly, some additional peaks with new masses were characterized, which may correspond to new fengycin classes. The isolates produced siderophores and indole-3- acetic acid phytohormone. The greenhouse experiment using a naturally infested soil with Sclerotonia sclerotiorum showed that the B. atrophaeus (6SEL) significantly increased the size of the chickpea plants and reduced the stem rot disease (P < 0.05). These results suggest that these isolates may be used further as bio-inoculants to improve crop systems