Keywords
phosphate solubilizing microorganisms
phosphatase activity
Box Behnken design
How to Cite
Abstract
Objective. To design a complex culture media for the production of biomass and acid phosphatases from phosphate-solubilizing bacteria isolated from soil. Materials and methods. Phosphate-solubilizing bacteria were isolated from oil palm crop soil samples and selected on SMRS1 agar, which were then assessed with antagonism tests to verify their aptitude to form a co-culture. A Box-Behnken experimental design was applied toevaluate the effect of each one of the culture media components on the production of biomass and phosphatase enzymes at a laboratory scale. Finally, microbial growth and enzyme production curves were carried out in order to determine their production times. Results. Five phosphate-solubilizing bacterial strains were isolated and three of them were selected based on their solubilization indices.These Gram negative strains with bacillus morphology were identified as A, B and C; their solubilization indices were 2.03, 2.12, and 2.83, respectively. According to the ANOVA analyses for the Box-Behnken design, the only factor which had a significant effect on the phosphatase activity (p<0.01) was hydrolyzed yeast, and the formulation that generated the highest biomass concentration and phosphatase activity (p<0.01) contained 10, 15 and 2.5 gL-1 of phosphoric rock, sucrose and hydrolyzed yeast, respectively. After 24 hours of incubation at 100 rpm, the highest values of biomass and phosphatase activity were obtained: 11.8 logarithmic units of CFU and 12.9 phosphatase units. Conclusion. We determined that the culture media based on phosphoric rock 10 gL-1, hydrolyzed yeast 2.5 gL-1 and commercial sucrose 15 gL-1 was ideal for the production of biomass and phosphatases by the strains evaluated; likewise, we proved that the hydrolyzed yeast was the only factor significantly influential for the production of phosphatases.
Key words: bio-inoculants, phosphate solubilizing microorganisms, phosphatase activity, Box Behnken design.
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