Recombinant Trypsin Inhibitor (ILTI) Production in Confirms Its Potential Anti-Biofilm Effect and Reveals an Anti-Tumoral Activity

Overview

Journal Microorganisms

Specialty Microbiology

Date 2018 May 2

PMID 29710773

Citations 2

Authors

Fabio C Carneiro

Simone S Weber

Osmar N Silva

Ana Cristina Jacobowski

Marcelo H S Ramada

Maria L R Macedo

Octavio L Franco

Nadia S Parachin

Affiliations

Soon will be listed here.

Abstract

Protease inhibitors have a broad biotechnological application ranging from medical drugs to anti-microbial agents. The trypsin inhibitor (ILTI) previously showed a great in vitro inhibitory effect under the adherence of species, being a strong candidate for use as an anti-biofilm agent. Nevertheless, this is found in small quantities in its sources, which impairs its utilization at an industrial scale. Within this context, heterologous production using recombinant microorganisms is one of the best options to scale up the recombinant protein production. Thus, this work aimed at utilizing to produce recombinant ILTI. For this, the vector pPIC9K+ILTI was constructed and inserted into the genome of the yeast , strain GS115. The protein expression was highest after 48 h using methanol 1%. A matrix-assisted laser desorption ionization⁻time-of-flight (MALDI⁻TOF) analysis was performed to confirm the production of the recombinant ILTI and its activity was investigated trough inhibitory assays using the synthetic substrate Nα-Benzoyl-D,L-arginine p-nitroanilide hydrochloride (BAPNA). Finally, recombinant ILTI (rILTI) was used in assays, showing that there was no significant difference between native and recombinant ILTI in its inhibitory activity in biofilm formation. Anti-tumor assay against Ehrlich ascites tumor (EAT) cells showed that rILTI has a potential anti-tumoral effect, showing the same effect as Melittin when incubated for 48 h in concentrations above 25 µg/mL. All together the results suggests broad applications for rILTI.

Citing Articles

A Novel Kunitz Trypsin Inhibitor from Seeds Exhibits Antibiofilm Properties against Pathogenic Yeasts.

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Special Issue: Recombinant Protein Expression in Microorganisms.

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