Production of Canecystatin in transgenic sugarcane vacuoles

• Grant number: 17/16118-1
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: December 01, 2017
• End date: March 31, 2021
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Flavio Henrique da Silva
• Grantee: Priscila Yumi Tanaka Shibao
• Host Institution: Centro de Ciências Biológicas e da Saúde (CCBS). Universidade Federal de São Carlos (UFSCAR). São Carlos , SP, Brazil
• Associated scholarship(s): 18/20051-2 - Production of a sugarcane cystatin in Nicotiana benthamiana, BE.EP.DR

Abstract

One of the major challenges in Brazilian sugarcane production is the weevil Sphenophorus levis. The major digestive proteins of S. levis belong to cysteine peptidases (cathepsins L -like). Sugarcane, on the other hand, presents endogenous defense by the production of phytocystatins, cysteine peptidases inhibitors. Based on sugarcane EST library (SUCEST), Laboratory of Molecular Biology produced and characterized several of these inhibitors (Cane-CPI-1 to Cane-CPI-6). CaneCPI-5 demonstrated high inhibitory capability against Sl-CathL, the major cathepsin produced by S. levis, besides inhibiting other cathepsins. In addition, previous studies carried out in our lab revealed that CaneCPI-5 is acid resistant and strongly binds to dental enamel, presenting commercial application for teeth protection against erosion (Patent deposited). Therefore, it is inevitable to obtain a transgenic lineage of sugarcane overexpressing CaneCPI-5 and directing this protein to lytic vacuoles, which will allow its accumulation and recovery from the sugarcane juice. A transgenic lineage with these characteristics should be more resistant to the weevil and other beetles that use cysteine peptidases as digestive enzymes by concentrating the protein, in addition to facilitate its purification. The present project aims in the production of caneCPI-5 in the vacuole of transgenic sugarcane obtained by biolistic bombardment and Agrobacterium tumenfaciens transformation, transgenic characterization, pest defiance and obtaining high yield of purified protein from the sugarcane juice.

Scientific publications

(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)

TANAKA SHIBAO, PRISCILA YUMI; SANTOS-JUNIOR, CELIO DIAS; SANTIAGO, ADELITA CAROLINA; MOHAN, CHAKRAVARTHI; MIGUEL, MARIANA CARDOSO; TOYAMA, DANYELLE; SANCHES VIEIRA, MARCOS ANTONIO; NARAYANAN, SUBRAMONIAN; FIGUEIRA, ANTONIO; CARMONA, ADRIANA K.; et al. Sugarcane cystatins: From discovery to biotechnological applications. International Journal of Biological Macromolecules, v. 167, p. 676-686, JAN 15 2021. (17/16118-1, 19/11683-8, 15/10855-9, 18/20051-2)

SHIBAO, PRISCILA YUMI TANAKA; FERRO, MILENE; DE PAULA, FERNANDO FONSECA PEREIRA; LIMA, BRUNO SALATA; HENRIQUE-SILVA, FLAVIO. Identification and Functional Analysis of a Pseudo-Cysteine Protease from the Midgut Transcriptome of Sphenophorus levis. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, v. 22, n. 21, NOV 2021. (17/16118-1, 13/11641-7)