Optimization of Penicillin G Acylase Immobilization Process by Surface Response Methodology Using Central Composite Design

Atyabi Seyed Mohammad,Salimi Mona,Hossein Nemati,Norouzian Dariush,Akbarzadeh Azim,Sedigeh Hatami Gigloo,Momen Seyed Bahman

doi:10.4236/am.2013.41012

Abstract

Penicillin G acylase was immobilized onto superparamagnetite iron oxide nanoparticles employing response surface methodology through central composite design. Polynomial quadratic model was selected as a model. The value of the determination coefficient (R2) calculated from the quadratic regression model was 0.845, while the value of the adjusted (R2) was 0.74. The regression analysis of the data showed that the quadratic model selected were appropriate thereby enzyme concentration (A), reaction temperature (D), enzyme concentration* reaction temperature (AD), quadratics enzyme concentration (A2) and reaction temperature (D2) were found to be significant factors in immobilization process of penicillin G acylase.

Highlights

The regression analysis of the data showed that the quadratic model selected were appropriate thereby enzyme concentration (A), reaction temperature (D), enzyme concentration* reaction temperature (AD), quadratics enzyme concentration (A2) and reaction temperature (D2) were found to be significant factors in immobilization process of penicillin G acylase
Penicillin G acylase is a biocatalyst produced by bacteria as well as fungi in order to acylate/deacylate penicillin G to its constituent 6-aminopenicilanic acid and phenyl acetic acid or vice versa which is termed as hydrolysis (H) and synthesis (S) respectively, (Figure 1)
Penicillin acylase activity was determined as reported by Norouzian et al [5], measuring the amount of 6-aminopenicilanic acid (6-APA) formed at 35 ̊C ± 1 ̊C, employing 2% (w/v) benzyl penicillin prepared in 0.1 M phosphate buffer pH 7.5

Summary

Introduction

Penicillin G acylase is a biocatalyst produced by bacteria as well as fungi in order to acylate/deacylate penicillin G to its constituent 6-aminopenicilanic acid and phenyl acetic acid or vice versa which is termed as hydrolysis (H) and synthesis (S) respectively, (Figure 1). Penicillin acylase in its soluble form is unstable, cannot be separated from the reaction mixture which will add to the production cost of the final product such a commercially, industrially important biocatalyst has been immobilized by different techniques employing various supports by many investigators [1,2,3,4,5,6,7]. Investigators are trying to employ technique and reagents which will improve the kinetic properties of the enzyme under immobilization process as compared to its soluble counterpart [7,8,9,10,11]. In. this article, we have attempted to optimize the immobilization process of penicillin G acylase onto super para magnetite iron oxide nanoparticles by considering factors such as, enzyme concentration, weight of nanoparticle, concentration of polyethyleneimine, temperature and time of reaction using surface response methodology. In RSM, the main effects and interactions between various factors, each at different levels can be simultaneously studied [12]

Materials

Penicillin G Acylase Activity

Immobilization of Penicillin G Acylase

Statistical Analysis

Results and Discussion

Design Points