Overexpression of the plg1 gene encoding pectin lyase in Penicillium griseoroseum.
The pectin lyase (PL) is an industrially important enzyme since it is used for maceration and clarification in the process of fruit juice production in food industries. In order to increase the yields of pectin lyase we cloned the plg1 (pectin lyase 1) from Penicillium griseoroseum gene under the control of the strong constitutive promoter of the glyceraldehyde-3-phosphate dehydrogenase gene (gpdA) and the terminator region of the tryptophan synthetase (trpC) gene from Aspergillus nidulans (plasmid pAN52-Plg1) and transformed this construct into the P. griseoroseum strain PG63. One of the pAN52-Plg1 multi-copy transformants (strain 105) grown in culture medium containing glucose or sugar cane juice showed PL activities of 4,804 or 5,202 U ml(-1) respectively, which represented 57- and 132-fold increases. In addition, the apparent specific activity of PL produced by this strain was much higher than the one observed for a commercial pectinase preparation. Evaluation of the extracellular proteins in the culture supernatant of strain 105 by SDS-PAGE showed the presence of a clear and strong band of approximately 40 kDa that probably corresponds to PL. The enzyme yields reported here demonstrate that the system we developed is able to express pectin lyase at levels comparable to, or exceeding, previously reported data.
Cardoso
Patrícia Gomes
PG
Departamento de Biologia/Setor de Microbiologia, Universidade Federal de Lavras, Lavras, MG, Brazil.
Ribeiro
João Batista
JB
Teixeira
Janaina Aparecida
JA
de Queiroz
Marisa Vieira
MV
de Araújo
Elza Fernandes
EF
D001233:Aspergillus nidulans / Q000235:genetics; D003470:Culture Media; D004274:DNA, Recombinant; D004591:Electrophoresis, Polyacrylamide Gel; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D025282:Mycelium; D030781:Organisms, Genetically Modified / Q000235:genetics; D010407:Penicillium / Q000201:enzymology* / Q000235:genetics*; D010957:Plasmids; D011133:Polysaccharide-Lyases / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic / Q000235:genetics; D013728:Terminator Regions, Genetic; D014170:Transformation, Genetic
D003470:Culture Media; D004274:DNA, Recombinant; D011133:Polysaccharide-Lyases; C027349:pectin lyase