Expression of an Erwinia pectate lyase in three species of Aspergillus.
Transgenic filamentous fungi of the species Aspergillus niger, A. nidulans and A. awamori expressing and secreting Erwinia carotovora subsp. atroseptica pectate lyase 3 (PL3) were generated. Correct processing of the pre-enzyme was achieved using the A. niger pectin lyase A (PEL A) signal peptide. With the prepro-peptide of A. niger polygalacturonase II, secreted enzymes still possessed the 6- aa pro-sequence, indicating the importance of the conformation of the precursor protein for correct cleavage of the signal sequence. PL3 expression was markedly increased in media optimized for limited protease activity, and reached 0.4, 0.8 and 2.0 mg/l for expression in A. niger, A. awamori and A. nidulans, respectively. Glycans attached to the PL3 enzymes exhibited species-specific differences, and an increase of molecular mass coincided with reduced specific activities of the enzymes.
Bartling
S
S
Department of Physiology, Carlsberg Laboratory, Gamle Carlsberg Vej 10, DK-2500 Copenhagen, Denmark.
van den Hombergh
J P
JP
Olsen
O
O
von Wettstein
D
D
Visser
J
J
D000595:Amino Acid Sequence; D001230:Aspergillus / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D004885:Erwinia / Q000201:enzymology* / Q000235:genetics; D015870:Gene Expression; D008969:Molecular Sequence Data; D011133:Polysaccharide-Lyases / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013045:Species Specificity; D014170:Transformation, Genetic
D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D011133:Polysaccharide-Lyases; C023911:pectate lyase