Immobilization in the presence of Triton X-100: modifications in activity and thermostability of Geobacillus thermoleovorans CCR11 lipase.
A partially purified lipase produced by the thermophile Geobacillus thermoleovorans CCR11 was immobilized by adsorption on porous polypropylene (Accurel EP-100) in the presence and absence of 0.1% Triton X-100. Lipase production was induced in a 2.5% high oleic safflower oil medium and the enzyme was partially purified by diafiltration (co. 500,000 Da). Immobilization conditions were established at 25 degrees C, pH 6, and a protein concentration of 0.9 mg/mL in the presence and absence of 0.1% Triton X-100. Immobilization increased enzyme thermostability but there was no change in neither the optimum pH nor in pH resistance irrelevant to the presence of the detergent during immobilization. Immobilization with or without Triton X-100 allowed the reuse of the lipase preparation for 11 and 8 cycles, respectively. There was a significant difference between residual activity of immobilized and soluble enzyme after 36 days of storage at 4 degrees C (P < 0.05). With respect to chain length specificity, the immobilized lipase showed less activity over short chain esters than the soluble lipase. The immobilized lipase showed good resistance to desorption with phosphate buffer and NaCl; minor loses with detergents were observed (less than 50% with Triton X-100 and Tween-80), but activity was completely lost with SDS. Immobilization of G. thermoleovorans CCR11 lipase in porous polypropylene is a simple and easy method to obtain a biocatalyst with increased stability, improved performance, with the possibility for re-use, and therefore an interesting potential use in commercial conditions.
Sánchez-Otero
M Guadalupe
MG
Instituto Tecnológico de Veracruz, Unidad de Investigación y Desarrollo en Alimentos, Veracruz, Mexico.
Valerio-Alfaro
Gerardo
G
García-Galindo
Hugo S
HS
Oliart-Ros
Rosa María
RM
D000327:Adsorption; D001406:Bacillaceae / Q000201:enzymology*; D003902:Detergents / Q000737:chemistry; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000378:metabolism*; D017830:Octoxynol / Q000737:chemistry*; D011126:Polypropylenes; D013379:Substrate Specificity; D013696:Temperature
D003902:Detergents; D004800:Enzymes, Immobilized; D011126:Polypropylenes; D017830:Octoxynol; D008049:Lipase