TY - JOUR
T1 - Expression of a Cutinase of Moniliophthora roreri with Polyester and PET-Plastic Residues Degradation Activity
AU - Vázquez-Alcántara, Laura
AU - Oliart-Ros, Rosa María
AU - García-Bórquez, Arturo
AU - Peña-Montes, Carolina
N1 - Publisher Copyright:
© 2021 Vázquez-Alcántara et al.
PY - 2021/12
Y1 - 2021/12
N2 - Cutinases are enzymes produced by phytopathogenic fungi like Moniliophthora roreri. The three genome-located cutinase genes of M. roreri were amplified from cDNA of fungi growing in different induction culture media for cutinase production. The mrcut1 gene was expressed in the presence of a cacao cuticle, while the mrcut2 and mrcut3 genes were expressed when an apple cuticle was used as the inducer. The sequences of all genes were obtained and analyzed by bioinformatics tools to determine the presence of signal peptides, introns, glycosylation, and regulatory sequences. Also, the theoretical molecular weight and pI were obtained and experimentally confirmed. Finally, cutinase 1 from M. roreri (MRCUT1) was selected for heterologous expression in Escherichia coli. Successful overexpression of MRCUT1 was observed with the highest enzyme activity of 34,036 U/mg under the assay conditions at 40°C and pH 8. Furthermore, the degradation of different synthetic polyesters was evaluated; after 21 days, 59% of polyethylene succinate (PES), 43% of polycaprolactone (PCL), and 31% of polyethylene terephthalate (PET) from plastic residues were degraded.
AB - Cutinases are enzymes produced by phytopathogenic fungi like Moniliophthora roreri. The three genome-located cutinase genes of M. roreri were amplified from cDNA of fungi growing in different induction culture media for cutinase production. The mrcut1 gene was expressed in the presence of a cacao cuticle, while the mrcut2 and mrcut3 genes were expressed when an apple cuticle was used as the inducer. The sequences of all genes were obtained and analyzed by bioinformatics tools to determine the presence of signal peptides, introns, glycosylation, and regulatory sequences. Also, the theoretical molecular weight and pI were obtained and experimentally confirmed. Finally, cutinase 1 from M. roreri (MRCUT1) was selected for heterologous expression in Escherichia coli. Successful overexpression of MRCUT1 was observed with the highest enzyme activity of 34,036 U/mg under the assay conditions at 40°C and pH 8. Furthermore, the degradation of different synthetic polyesters was evaluated; after 21 days, 59% of polyethylene succinate (PES), 43% of polycaprolactone (PCL), and 31% of polyethylene terephthalate (PET) from plastic residues were degraded.
KW - Cutinase
KW - Moniliophthora roreri
KW - Polyester degradation
UR - http://www.scopus.com/inward/record.url?scp=85121678278&partnerID=8YFLogxK
U2 - 10.1128/Spectrum.00976-21
DO - 10.1128/Spectrum.00976-21
M3 - Artículo
C2 - 34730414
AN - SCOPUS:85121678278
SN - 2165-0497
VL - 9
JO - Microbiology Spectrum
JF - Microbiology Spectrum
IS - 3
M1 - e00976-21
ER -