Cloning and expression of animal toxins of biotechnological interest

• Grant number: 12/14996-8
• Support Opportunities: Regular Research Grants
• Start date: October 01, 2012
• End date: September 30, 2014
• Field of knowledge: Health Sciences - Pharmacy - Toxicological Analysis
• Principal Investigator: Eliane Candiani Arantes Braga
• Grantee: Eliane Candiani Arantes Braga
• Host Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated researchers: Flavio Henrique da Silva ; Ludier Kesser Santos Silva ; Renata Santos Rodrigues ; Simone Kashima Haddad

Abstract

Currently, the interest in the studying chemical and functional characteristics of toxins isolated from animal venoms is not due solely to its relevance in the envenoming, but also for their potential use as valuable research tools. The hyaluronidases from scorpion and snake venoms are enzymes that catalyze the breakdown of hyaluronan, a major component of the extracellular matrix and act as "scattering factor" for accelerating the absorption and diffusion of the venom by tissues of the victims, contributing to the local or systemic envenoming. Although hyaluronidases have been extensively used in various fields of medicine, the number of published studies about this enzyme is reduced, due to its lack of toxicity and the difficulties in purifying of the enzyme from venoms. By the other hand, snake venom serine proteases are considered promising in the development of drugs for treatment of certain hemostatic disorders by acting on specific points of the coagulation cascade, mimicking natural regulatory components. These enzymes act mainly on plasma proteins, platelet aggregation, blood pressure and complement and nervous system. Despite the diversity of pathophysiological actions and pharmacological potential presented by animal toxins, there is a discrepancy between the enormous number of proteins isolated and characterized and the amount of proteins that have become an effective drug, which is due to limitations in obtaining toxins in sufficient quantities to a detailed structural and functional characterization and the impossibility of manipulation of the native molecule. Thus, the heterologous expression of proteins is an alternative for obtaining toxins in laboratory scale for the study of its enzymatic and functional activities. Therefore, this project aims to clone and express hyaluronidase from T. serrulatus and serine proteases from B. pauloensis and C. d. collilineatus venom gland in a heterologous system, which consists of a viable alternative for obtaining them in sufficient quantities to study their enzymatic and biological activities. Furthermore, the comparative study of the toxins in native and recombinant forms can generate prospects for the production of these toxins on an industrial scale, aiming its therapeutic and pharmacological application. (AU)

Scientific publications

(6)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

AMORIM, FERNANDA GOBBI; LONGHIM, HELOISA TAVONI; COLOGNA, CAMILA TAKENO; DEGUELDRE, MICHEL; DE PAUW, EDWIN; QUINTON, LOIC; ARANTES, ELIANE CANDIANI. Proteome of fraction from Tityus serrulatus venom reveals new enzymes and toxins. Journal of Venomous Animals and Toxins including Tropical Diseases, v. 25, APR 18 2019. (14/07824-1, 15/17466-8, 12/14996-8, 13/26083-0, 11/23236-4, 14/22959-0, 13/26200-6)

AMORIM, FERNANDA GOBBI; BOLDRINI-FRANCA, JOHARA; FIGUEIREDO BORDON, KARLA DE CASTRO; CARDOSO, IARA AIME; DE PAUW, EDWIN; QUINTON, LOUC; KASHIMA, SIMONE; ARANTES, ELIANE CANDIANI. Heterologous expression of rTsHyal-1: the first recombinant hyaluronidase of scorpion venom produced in Pichia pastoris system. Applied Microbiology and Biotechnology, v. 102, n. 7, p. 3145-3158, APR 2018. (11/12317-3, 13/26083-0, 12/14996-8)

AMORIM, FERNANDA GOBBI; CORDEIRO, FRANCIELLE ALMEIDA; PINHEIRO-JUNIOR, ERNESTO LOPES; BOLDRINI-FRANCA, JOHARA; ARANTES, ELIANE CANDIANI. Microbial production of toxins from the scorpion venom: properties and applications. Applied Microbiology and Biotechnology, v. 102, n. 15, p. 6319-6331, AUG 2018. (16/04761-4, 14/16182-3, 11/12317-3, 12/14996-8)

BOLDRINI-FRANCA, JOHARA; PINHEIRO-JUNIOR, ERNESTO LOPES; ARANTES, ELIANE CANDIANI. Functional and biological insights of rCollinein-1, a recombinant serine protease from Crotalus durissus collilineatus. Journal of Venomous Animals and Toxins including Tropical Diseases, v. 25, APR 8 2019. (16/04761-4, 14/16182-3, 12/14996-8)

DE OLIVEIRA, ISADORA SOUSA; PUCCA, MANUELA BERTO; SAMPAIO, SUELY VILELA; ARANTES, ELIANE CANDIANI. Antivenomic approach of different Crotalus durissus collilineatus venoms. Journal of Venomous Animals and Toxins including Tropical Diseases, v. 24, NOV 26 2018. (11/23236-4, 12/14996-8)

DE OLIVEIRA, ISADORA SOUSA; MANZINI, RAFAELLA VARZONI; FERREIRA, ISABELA GOBBO; CARDOSO, IARA AIME; FIGUEIREDO BORDON, KARLA DE CASTRO; THOMAZELA MACHADO, ANA RITA; GREGGI ANTUNES, LUSANIA MARIA; ROSA, JOSE CESAR; ARANTES, ELIANE CANDIANI. Cell migration inhibition activity of a non-RGD disintegrin from Crotalus durissus collilineatus venom. Journal of Venomous Animals and Toxins including Tropical Diseases, v. 24, OCT 20 2018. (16/11578-1, 11/23236-4, 12/14996-8, 17/03580-9)