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Master's Dissertation
DOI
https://doi.org/10.11606/D.75.2009.tde-25082009-164952
Document
Author
Full name
Denis Massucatto
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Carlos, 2009
Supervisor
Committee
Gehlen, Marcelo Henrique (President)
Porto, Andre Luiz Meleiro
Stevani, Cassius Vinicius
Title in Portuguese
Investigação da cinética de lipase através de espectroscopia de fluorescência
Keywords in Portuguese
fluorescência
hidrólise enzimática
lipase
quinizarina
reação em duas etapas sequenciais
Abstract in Portuguese
As enzimas constituem parte essencial à vida, sendo catalisadores capazes de aumentar em várias ordens de grandeza a velocidade da reação. As lipases, que catalisam a hidrólise de triacilglicerídeos, são enzimas versáteis e de fácil obtenção e por isso estão presentes em diversos setores da indústria e também constituem tema de estudo no meio acadêmico. Tem-se como os objetivos principais deste trabalho a elaboração de um método para se avaliar a cinética hidrólise de lipase suportada por intermédio de espectroscopia de fluorescência. O sistema utilizado foi o DAQ (diaceto-quinizarina) / quinizarina. Este sistema é propício, pois a DAQ, que não é fluorescente, quando hidrolisada resulta na quinizarina, altamente fluorescente. Para se tratar a cinética de hidrólise foi desenvolvido um modelo considerando-se duas etapas seqüenciais de hidrólise, uma vez que a enzima catalisa a quebra de apenas um grupo éster por ciclo catalítico. O modelo mostrou-se bastante fiel no ajuste dos dados experimentais obtidos, sendo possível obter a constante cinética envolvida no processo global de hidrólise, que constitui uma combinação linear das constantes cinéticas dos processos elementares.
Title in English
Investigation of lipase kinetics through fluorescence spectroscopy
Keywords in English
enzymatic hydrolisys
fluorescence
lipase
quinizarin
two-step sequential reaction
Abstract in English
Enzymes are essential to life and are capable to increase in many orders of magnitude the velocity of reaction. Lipases, that catalyses the hydrolysis of triacylglyceride, are versatile enzymes and easy to obtain and are present in many industry segments and also constitutes academic studies. The objective of this work is the elaboration of a method to evaluate the hydrolysis kinetic of immobilized lipase through fluorescence spectroscopy. The system used was DAQ (diacetate-quinizarin) / quinizarin. This system is auspicious because DAQ, not-fluorescent, when hydrolyzed results quinizarin, highly fluorescent. To treat the hydrolysis kinetic, a model that consider a twostep sequential reaction were developed, once the enzymes breaks only one ester in each catalytic cycle. The model agreed with the experimental data, and it was possible to obtain the global kinetic constant, that is a linear combination of the elementary constants
 
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DenisMassucattoR.pdf (2.91 Mbytes)
Publishing Date
2009-08-26
 
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