Process Biotechnology
  Biotechnology Industry
Electronic Journal of Biotechnology ISSN: 0717-3458 Vol. 14 No. 3, Issue of May 15, 2011
© 2011 by Pontificia Universidad Católica de Valparaíso -- Chile Received October 20, 2010 / Accepted January 27, 2011
DOI: 10.2225/vol14-issue3-fulltext-1  

Influence of the pH of glutaraldehyde and the use of dextran aldehyde on the preparation of cross-linked enzyme aggregates (CLEAs) of lipase from Burkholderia cepacia

Eduardo Caballero Valdés*1 · Lorena Wilson Soto1 · Germán Aroca Arcaya1

1Facultad de Ingeniería, Escuela de Ingeniería Bioquímica, Pontificia Universidad Católica de Valparaíso, Valparaíso, Chile

*Corresponding author: 

Financial support: This project was financed by a scholarship granted by CONICYT, Chile.

Keywords: albumin, dextran aldehyde, glutaraldehyde, immobilisation, lipase.


The preparation of cross-linked enzyme aggregates (CLEAs) of lipase has been a challenge due the low amount of lysine residues that lipases have on their surface. The results show that CLEAs prepared using dextran aldehyde (100-200KDa) have a higher hydrolysis activity and particle size (activities between 3186 ± 21 U/g of CLEA and 4800 ± 30 U/g of CLEA and particle sizes between 52.6 ± 18.7 µm and 126.2 ± 53.5 µm) than CLEAs prepared with glutaraldehyde (0.1 KDa) (activities between 894 ± 16 U/g of CLEA and 2874 ± 20 U/g of CLEA and particle sizes between 21.2 ± 5.1 µm and 83.4 ± 24.9 µm); Thermal stability assays of bioctalysts at 60ºC at pH 7.0 using phosphate buffer 25 mM showed that CLEAs prepared with dextran aldehyde have lower residual activity after 50 hrs (maximum residual activity of 46.8% in the CLEA) than CLEAs prepared with glutaraldehyde (maximum residual activity of 70.2% in CLEA). When considering hydrolysis activity, thermal stability and residual activity of CLEAs as a criteria for selecting the best preparation conditions, it has been found that the best condition for CLEAs preparation are to use glutaraldehyde as cross-linking reagent at pH 9.5, at a concentration of 3.5 g/l, and an enzyme/albumin ratio of 15.