Response Surface Methodology Analysis of

Biodegradation of Acrylonitrile in Bioreactor

Qi Wang

School of Life and Environmental Sciences, Wenzhou University, 325035, China

victor527@126.com

Abstract. Response surface methodology (RSM) was used to study the

biodegradation effect of the various parameters namely, initial
acrylonitrile(AN) concentration, pH, temperature and to optimize the process
conditions for the maximum degradation of acrylonitrile by implementing the
Box-Behnken statistical design in bioreactor. The analysis of variance
(ANOVA) of the quadratic model demonstrates that the model was highly
significant. Using this methodology, the optimal values of pH, temperature,
℃
initial AN concentration were 6.54, 26.22 and 424.32mg/L, respectively. The
experiment results showed that the RSM based on Box-Behnken design was
useful for optimizing the biodegradation process.

Keywords: Response Surface Methodology, Acrylonitrile, Biodegradation.

1 Introduction
Acrylonitrile(AN) is an raw material that has been extensively used for manufacturing
acrylic fiber, synthetic chemistry and synthetic rubber. It is the priority pollutant in
many countries [1] including the United States, China and Germany for its toxicity.
AN wastewater, with high strength chemical oxygen demand (COD) and AN, was
treated by techniques including activated sludge treatment [2-4], wet air oxidized and
activated carbon treatment [5],ect. Treatment techniques of refractory biodegradation
organic pollutants are hot point to current water pollution control due to its serious
pollution and hazardousness to people's health in water, soil and sediments.
Response surface methodology (RSM), an experimental approach for finding the
optimum conditions, is an efficient graphical statistical technique to identify different
factors that produce the best operation or process. RSM was first used for optimizing
machining techniques [6]. Recently, it has been successfully applied to a different
process for achieving its optimization using RSM, which included bioremediation
studies [7, 8], enzyme optimum reactions[9-15], wastewater treatment progress[16-18],
production of activated carbon and xylose [19, 20], ect. Physicochemical methods have
high treatment costs and may cause secondary pollution; hence this study attempts to
investigate AN removal by the biological method. In the present study, response surface
methodology was used to study the biodegradation effect of the various parameters to
optimize the process conditions for the maximum degradation of acrylonitrile by
implementing the Box-Behnken statistical design in bioreactor.

H. Tan and M. Zhou (Eds.): CSE 2011, Part I, CCIS 201, pp. 31–36, 2011.
© Springer-Verlag Berlin Heidelberg 2011
32 Q. Wang

2 Materials and Methods

2.1 Materials

Original mixed bacteria were obtained from soil samples polluted by nitriles for long
period of time. Rice husk was supplied by Dalian food storage and transportation
industrial company, Dalian, China.

2.2 Microorganism Medium and Culture Conditions

Microorganisms were collected and accumulated in rice husk biofilm culture

℃
equipment. Cultures were maintained at 25 and 150 rpm for the duration mentioned
in the experiment design. More than 90% AN degradation after 24 h was obtained and
this indicated that the rice husk carrier for immobilized microbial could treat AN
wastewater at normal operational condition.

2.3 Analytical Methods

The AN concentration was analyzed by using pyridine colorimetric analysis method.

The pH was measured by glass microelectrode numerical pH-meter (Shanghai
Precision Instruments, China).

2.4 Design of Experiments by Response Surface Methodology

Response surface methodology is a combination of statistical and optimization

technique used to evaluate the relationship between a set of controllable experimental
factors and observed results. This optimization process involves three major steps: (i)
performing statistically designed experiments, (ii) estimating the coefficients in a
mathematical model, and (iii) predicting the response and checking the adequacy of
the model[21].
A design matrix comprising of 17 experimental runs was constructed. The non-
linear computer-generated quadratic equation model for predicting the optimal point
can be expressed according to Eq. (1):
where y is the measured response associated with each factor level combination; b0
is an intercept;b1 to b33 are regression coefficients computed from the observed
experimental values of y; and x1, x2 and x3 are the coded values levels of independent
variables. The terms x12, x13, x23 and xi (i=1, 2or 3) represent the interaction and
quadratic terms, respectively[22].
Box-Behnken design was specifically selected since it requires fewer runs than a
central composite design(CCD) in cases of three or four variables. Data were
processed using Design-Expert (version 7.0, Stat–Ease, Minneapolis, USA) program
including ANOVA to obtain the interaction between the process variables and the
response. The quality of the fit of polynomial model was expressed by the coefficient
of determination R2, and its statistical significance was checked. From three-
dimensional plots, the simultaneous interaction of two factors on the response was
studied.