Journal: Applied Microbiology and Biotechnology

Article Title: Recombinant Aspergillus β-galactosidases as a robust glycomic and biotechnological tool

doi: 10.1007/s00253-013-5192-3

Figure Lengend Snippet: Galactosidase phenogram and SDS-PAGE analysis of recombinant enzymes. a Protein alignment based on the ClustalW method was performed in MegAlign 7.2.1 (DNAstar Lasergene software package) using full-length protein sequences of galactosidases as obtained from the GenBank database. The phenogram was prepared using the MegAlign’s built-in phylogenetic tree function. The enzymes described in this study are indicated by a gray background (including the A. oryzae enzyme used for comparative purposes). The amino acid sequences with the following IDs were used: A. niger lacA ID XP_001389622, A. niger ID A2QL84, A. nidulans lacA ID XP_658360, A. nidulans lacB ID XP_658584, A. clavatus ID XP_001268843, A. fumigatus ID XP_752787, A. fumigatus ID XP_748360, A. oryzae ID XP_001727461, Penicilium sp. ID Q700S9, Trichoderma reesei CAD70669, Homo sapiens ID NP_000395, K. lactis XP_452194, K. lactis 3OB8_A, and Escherichia coli ID NP_414878. b SDS-PAGE (Tris–glycine; resolving gel: T12.5/C1, stacking gel: T5.7/C2.2) analysis of purified recombinant enzymes. Each galactosidase was obtained free from visible contaminations as judged by Coomassie Brilliant Blue staining. Purified recombinant proteins were incubated with (+) or without (−) PNGase F to verify whether they carry N -glycans. MW molecular weight in kilodaltons

Article Snippet: Fig. 1 Galactosidase phenogram and SDS-PAGE analysis of recombinant enzymes. a Protein alignment based on the ClustalW method was performed in MegAlign 7.2.1 (DNAstar Lasergene software package) using full-length protein sequences of galactosidases as obtained from the GenBank database.

Techniques: SDS Page, Recombinant, Software, Purification, Staining, Incubation, Molecular Weight