Statistical experimental methods for optimizing the cultivating conditions for Rhodococcus erythropolis

Abstract

Rhodococcus erythropolis was found to effectively degrade aflatoxin Bl produced by Aspergillus flavus and Aspergillus parasiticus. However, one problem of concern was the slow growth of this strain. In this study, Plackett–Burman design was used to select the most important variables, namely, temperature, pH, inoculum size, liquid volume, agitation speed and culture time that affected the growth of R. erythropolis. Central composite experimental design and response surface analysis were adopted to derive a statistical model for optimizing the culture conditions. From the obtained results, it can be concluded that the optimum parameters were: temperature, 15.3°C; pH, 5.56; inoculum size, 4%; liquid volume, 70 ml in 250 ml flask; agitation speed, 180 rpm; and culture time, 58.2 h. At this optimum point, the populations of the viable organisms could reach 10⁸ colony forming units (CFU)/ml, which was 100 times higher than that incubated under the initial conditions. After 58.2 h incubation in this optimum cultivating conditions, 53.9 ± 2.1% of aflatoxin B₁ was degraded, while only 20.6±1.4% of aflatoxin B₁ was degraded in the initial conditions.

Key words: Rhodococcus erythropolis, culture condition, optimization, Plackett–Burman design, central composite design, response surface methodology.