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Enzyme Research
Volume 2012, Article ID 329178, 12 pages
Research Article

Optimal Conditions for Continuous Immobilization of Pseudozyma hubeiensis (Strain HB85A) Lipase by Adsorption in a Packed-Bed Reactor by Response Surface Methodology

1Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Porto Alegre 91501-970, RS, Brazil
2Laboratório de Química de Proteínas, Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Goiás, Goiânia 74001-970, GO, Brazil

Received 15 June 2011; Revised 10 October 2011; Accepted 10 October 2011

Academic Editor: Jose Miguel Palomo

Copyright © 2012 Roberta Bussamara et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


This study aimed to develop an optimal continuous process for lipase immobilization in a bed reactor in order to investigate the possibility of large-scale production. An extracellular lipase of Pseudozyma hubeiensis (strain HB85A) was immobilized by adsorption onto a polystyrene-divinylbenzene support. Furthermore, response surface methodology (RSM) was employed to optimize enzyme immobilization and evaluate the optimum temperature and pH for free and immobilized enzyme. The optimal immobilization conditions observed were 150 min incubation time, pH 4.76, and an enzyme/support ratio of 1282 U/g support. Optimal activity temperature for free and immobilized enzyme was found to be 68°C and 52°C, respectively. Optimal activity pH for free and immobilized lipase was pH 4.6 and 6.0, respectively. Lipase immobilization resulted in improved enzyme stability in the presence of nonionic detergents, at high temperatures, at acidic and neutral pH, and at high concentrations of organic solvents such as 2-propanol, methanol, and acetone.