Immobilization of the LipC12 recombinant lipase obained by metagenomic bioprospection and its evaluation in reactions of interest in oleochemistry

BECKER LOBERMAYER K.; DE SOUZA MACHADO A.; MARQUEZ DE ALMEIDA J.; ALNOCH R.; TONETTO G.; SANCHEZ D.; MITCHELL D.; FERREIRA M.; KRIEGER N.

Florianopolis

Congreso; XIII ENZITEC - Seminário Brasileiro de Tecnologia Enzimática; 2018

Universidade Federal de Santa Catarina y Universidade Federal da Fronteira Sul

In previous work, we have reported the successful expression and characterization of a lipase (LipC12) from a metagenomic library derived from a fat-contaminated soil. LipC12 is a true lipase that shows high specific activities, 1722 U mg-1 against olive oil. In the present work, we immobilized LipC12 in different supports and evaluated its potential for applications in reactions of interest in the oleochemistry industry. LipC12 was immobilized on the supports Immobead 150 and Sepabeads, wich allow the covalent and hydrophobic bonding of the enzyme, respectively. Protein loading of 10 mg g-1 was evaluated for the immobilization and key parameters were determined: the immobilization efficiency, the hydrolytic and esterification activity. The immobilization efficiency of LipC12 on Immobead 150 (Im-LipC12) was 92 ± 1%, while in Sepabeads (Se-LipC12) was 87 ± 0%, after 8 h of the immobilization. Im-LipC12 and Se-LipC12 exhibited hydrolysis activity against olive oil of 131 ± 5 and 224 ± 7 U g-1, respectively. The esterification activity was 15 ± 1 U g-1 for Im-LipC12 and 26 ± 2 U g-1 for Se-LipC12, in the synthesis of ethyl oleate in n-hexane, giving a conversion of 100% in 6 h at 40 °C, for both preparations. This difference in hydrolysis and esterification activity can be explained by the different bonding of the enzyme in the support. In Sepabead, which is hydrophobic, LipC12 probably bonds to the support via lid, which keeps the enzyme in its open conformation allowing greater activity. In conclusion, the support in which the immobilization occurs by hydrophobic interaction, Sepabead, is a better support for the immobilization of LipC12, compared to Immobead 150. The high esterification of LipC12 immobilized on Sepabead fosters its application in reaction of interest in the oleochemistry industry, such as in biodiesel synthesis and the production of structured lipids.