Available online 6 December 2022

GDSL-like esterase/lipase proteins (GELPs) are enzymes that possess unique characteristics, mainly because they contain four invariable catalytic residues. Advances in the study of these proteins are interesting. The cloning and functional expression of a papaya esterase have not been reported. Therefore, in this work we evaluated the heterologous production of Carica papaya esterase CpEST in the yeast Komogataella phaffii (Pichia pastoris).

The cloning and expression of the protein was performed under the PAOX1 promoter, and productions of up to 43 AU/mL were achieved using residual glycerol from biodiesel in the batch phase and methanol for the induction phase. Enzyme activity assays determined that CpEST has a high preference for short-chain substrates (p-NP C4 and p-NP C8), and optimal activity conditions were observed at 30°C. The enzyme showed the highest stability to acetone, ethanol and tert-butanol solvents, retaining approximately 55% of its initial enzymatic activity after 1 h of exposure.

Cloning and functional expression of papaya CpEST esterase was achieved. During fermentation, the yeasts used as a carbon source residual glycerol from biodiesel production. Based on the results obtained from the characterization of the esterase, it was found that it has a high potential for use in the bioenergy and detergent industry.

Biodiesel residual glicerol

Bioenergy

Carica papaya

Circular economy

Fermentation

GDSL-like esterase/lipase proteins

Heterologous expression

Papaya esterase

Plant enzymes

Wastes recovery

© 2022 Published by Elsevier B.V. on behalf of Pontificia Universidad Católica de Valparaíso.