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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

A Collagenolytic Aspartic Protease from Thermomucor indicae-seudaticae Expressed in Escherichia coli and Pichia pastoris

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Author(s):
Pereira, Waldir Eduardo Simioni [1] ; da Silva, Ronivaldo Rodrigues [1] ; de Amo, Gabriela Salvador [2] ; Ruller, Roberto [3] ; Kishi, Luciano Takeshi [4] ; Boscolo, Mauricio [1] ; Gomes, Eleni [1] ; da Silva, Roberto [1]
Total Authors: 8
Affiliation:
[1] Univ Estadual Paulista Unesp, Inst Biociencias Letras & Ciencias Exatas Ibilce, Campus Sao Jose Rio Preto, Sao Paulo - Brazil
[2] Inst Fed Educ Ciencia & Tecnol Sao Paulo, Campus Catanduva, Catanduva, SP - Brazil
[3] Univ Fed Mato Grosso do Sul, Campo Grande, MS - Brazil
[4] Lab Nacl Computacao Cient, Petropolis, RJ - Brazil
Total Affiliations: 4
Document type: Journal article
Source: Applied Biochemistry and Biotechnology; v. 191, n. 3 FEB 2020.
Web of Science Citations: 1
Abstract

Proteases are produced by the most diverse microorganisms and have a wide spectrum of applications. However, the use of wild microorganisms, mainly fungi, for enzyme production has some drawbacks. They are subject to physiological instability due to metabolic adaptations, causing complications and impairments in the production process. Thus, the objective of this work was to promote the heterologous expression of a collagenolytic aspartic protease (ProTiN31) from Thermomucor indicae seudaticae in Escherichia coli and Pichia pastoris. The pET\_28a (+) and pPICZ alpha A vectors were synthesized containing the gene of the enzyme and transformed into E. coli and P. pastoris, respectively. The recombinant enzymes produced by E. coli and P. pastoris showed maximum activity at pH 5.0 and 50 degrees C, and pH 5.0 and 60 degrees C, respectively. The enzyme produced by P. pastoris showed better thermostability when compared to that produced by E. coli. Both enzymes were stable at pH 6.0 and 6.5 for 24 h at 4 degrees C, and sensitive to pepstatin A, beta-mercaptoethanol, and Hg2+. Comparing the commercial collagen hydrolysate (Artrogen duo/Brazil) and gelatin degradation using protease from P. pastoris, they showed similar peptide profiles. There are its potential applications in a wide array of industrial sectors that use collagenolytic enzymes. (AU)

FAPESP's process: 17/14629-9 - Heterologous expression of milk-clotting protease pruduced by Thrmomucor indicae-seudaticae N-31 in Escherichia coli and Pichia Pastoris
Grantee:Waldir Eduardo Simioni Pereira
Support type: Scholarships in Brazil - Master