Evaluation of activity of a commercial amylase under ultrasound-assisted irradiationby Matheus Souza, Ethiane T. Mezadri, Eduardo Zimmerman, Eloisa X. Leaes, Mariana M. Bassaco, Valéria Dal Prá, Edson Foletto, Adriano Cancellier, Lisiane M. Terra, Sérgio L. Jahn, Marcio A. Mazutti

Ultrasonics Sonochemistry

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Accepted 23 May 2012

Available online 1 June 2012

Amylase

Ultrasound

Activation energy wor tral in t the influence of treatment time as well as the substrate concentration on enzyme activity were also determined in the presence and absence of ultrasound irradiation. The results demonstrated that the showed activities about 3 times higher for temperatures up to 40 C in the presence of ultrasound. The nd has al trea [4–8] there are few works evaluating the influence of ultrasound on enzyme characteristics [3,9–11].

The use of ultrasound to increase the activity/stability of enzymes, as well as, to reduce the mass transfer limitations in the process makes this treatment a promising technology for the conversion of raw starch under mild conditions using amylases, with very recent industrial application for the production of bioethanol the effects of ultrasound on the activity of STARGEN™ 002 that is an optimized blend of amylases produced by genetically modified strains of trichoderma reesei modified strains of Trichoderma reesei to hydrolyze raw starch. A central composite design was carried out to evaluate the effects of temperature and pH on enzyme activity. In addition, several experiments were carried out to determine the activation energy, the influence of treatment time as well as the substrate concentration on enzyme activity. All experimental conditions presented in this manuscript were carried out in the presence and absence of ultrasound irradiation. ⇑ Corresponding author. Tel.: +55 55 3220 9592.

Ultrasonics Sonochemistry 20 (2013) 89–94

Contents lists available at o .e lE-mail address: mazutti@ufsm.br (M.A. Mazutti).technology processes [1,2]. The chemical and physical effects of ultrasound arise from the cavitational collapse, which produces extreme conditions locally and thus induces the formation of chemical species not easily attained under conventional conditions, driving a particular reactivity. When cavitation bubbles collapse near the phase boundary of two immiscible liquids, the resultant shock wave can provide a very efficient stirring/mixing of the layers. As a consequence, the cavitation phenomenon can increase reaction rates and hence may become a useful tool in enzymatic reactions [3]. Although the application of ultrasound irradiation to enzymatic reactions is being increasingly explored

The amylolytic complex comprises groups of enzymes that act synergistically to break down both amylose (linear a-1,4-linked glucose units) and amylopectin (glucose-based homopolymer with linear chains and a-1,6-linked branches) to glucose [12–14]. Apar et al. [7] investigated the effect of ultrasonic energy on the hydrolysis of corn, rice, and wheat starch by using the a-amylase as well as the effects of sonifier operation variables such as duty cycle and acoustic power rate on the stability of a-amylase. From our knowledge, the effects of ultrasound on the characteristics of amylases have not been reported in literature until this moment.

Based on these aspects, this work is focused on the evaluation ofActivity 1. Introduction

In the last decade power ultrasou tive processing option to convention1350-4177/$ - see front matter  2012 Elsevier B.V. A http://dx.doi.org/10.1016/j.ultsonch.2012.05.012pH negatively affected the activity in the presence of ultrasound, whereas in the absence a positive effect was verified. The ultrasound irradiation is a promising technology to be used in enzymatic reaction due to its positive effects on enzyme activity.  2012 Elsevier B.V. All rights reserved. emerged as an alternatments in food and biofrom starch-rich agroindustrial residues. Amylases represent one of the most important groups of industrial enzymes worldwide.

They act in the hydrolysis of starch and present several applications in the food, pulp and paper, textile, and bioethanol industries.Keywords: effect of temperature was less pronounced in the presence of ultrasound, resulting in a decreasing of about 80% in the activation energy in comparison with the value obtained in the absence. The enzymeEvaluation of activity of a commercial am irradiation

Matheus Souza, Ethiane T. Mezadri, Eduardo Zimme

Valéria Dal Prá, Edson Foletto, Adriano Cancellier, Li

Department of Chemical Engineering, Federal University of Santa Maria, Av. Roraima, 1 a r t i c l e i n f o

Article history:

Received 28 February 2012

Received in revised form 24 April 2012 a b s t r a c t

The main objective of this commercial amylase. A cen pH on the enzyme activity

Ultrasonics S journal homepage: wwwll rights reserved.lase under ultrasound-assisted an, Eloisa X. Leaes, Mariana M. Bassaco, ne M. Terra, Sérgio L. Jahn, Marcio A. Mazutti ⇑

Santa Maria 97105-900, Brazil k was to evaluate the effects of ultrasound irradiation on the activity of a composite design was carried out to assess the effects of temperature and he presence and absence of ultrasound irradiation. The activation energy,

SciVerse ScienceDirect nochemistry sevier .com/locate /u l tson 2. Material and methods 2.1. Materials

The enzyme used in this work was STARGEN™ 002, gently donated by Genencor (Brazil). This enzyme is an optimized blend of amylases derived from genetically modified strains of T. reesei and it is used to cold hydrolysis of raw starch. The soluble potato starch and 3,5 dinitrosalicylic acid were purchase from Vetec and were used as received. activity was determined as described below. For the sample treated in the presence of ultrasound irradiation the activity also was determined in the ultrasound bath, whereas for the enzyme treated in the absence of ultrasound the activity was determined in the water bath. 2.6. Effect of ultrasound on the kinetics parameters of the Michaelis–

Menten equation

Enzyme assays containing 1 mL of commercial enzyme diluted d in ity h (U 90 M. Souza et al. / Ultrasonics Sonochemistry 20 (2013) 89–942.2. Equipment

Experiments were carried out in a reactor with thermostatic water bath (temperature accuracy of ±1.0 C). The experimental setup consists of an ultrasonic bath (Unique Inc., model USC 1800A, Brazil, BR) equipped with a transducer having longitudinal vibrations. The ultrasonic unit has an operating frequency of 40 kHz and a maximum-rated electrical power output of 132 W.