Next Generation Sequencing Identifies Five Major Classes of Potentially Therapeutic Enzymes Secreted by Lucilia sericata Medical Maggots.
Lucilia sericata larvae are used as an alternative treatment for recalcitrant and chronic wounds. Their excretions/secretions contain molecules that facilitate tissue debridement, disinfect, or accelerate wound healing and have therefore been recognized as a potential source of novel therapeutic compounds. Among the substances present in excretions/secretions various peptidase activities promoting the wound healing processes have been detected but the peptidases responsible for these activities remain mostly unidentified. To explore these enzymes we applied next generation sequencing to analyze the transcriptomes of different maggot tissues (salivary glands, gut, and crop) associated with the production of excretions/secretions and/or with digestion as well as the rest of the larval body. As a result we obtained more than 123.8 million paired-end reads, which were assembled de novo using Trinity and Oases assemblers, yielding 41,421 contigs with an N50 contig length of 2.22 kb and a total length of 67.79 Mb. BLASTp analysis against the MEROPS database identified 1729 contigs in 577 clusters encoding five peptidase classes (serine, cysteine, aspartic, threonine, and metallopeptidases), which were assigned to 26 clans, 48 families, and 185 peptidase species. The individual enzymes were differentially expressed among maggot tissues and included peptidase activities related to the therapeutic effects of maggot excretions/secretions.
Department of Bioresources, Fraunhofer Institute for Molecular Biology and Applied Ecology, Winchesterstraße 2, 35394 Giessen, Germany.
Zdeněk
Z
Franta
Department of Entomology, Max Planck Institute for Chemical Ecology, Hans-Knöll-Straße 8, 07745 Jena, Germany.
Heiko
H
Vogel
Department of Bioresources, Fraunhofer Institute for Molecular Biology and Applied Ecology, Winchesterstraße 2, 35394 Giessen, Germany.
Rüdiger
R
Lehmann
Justus-Liebig-University of Giessen, Bioinformatics and System Biology, Heinrich-Buff-Ring 58, 35392 Giessen, Germany.
Oliver
O
Rupp
Justus-Liebig-University of Giessen, Bioinformatics and System Biology, Heinrich-Buff-Ring 58, 35392 Giessen, Germany.
Alexander
A
Goesmann
Department of Bioresources, Fraunhofer Institute for Molecular Biology and Applied Ecology, Winchesterstraße 2, 35394 Giessen, Germany; Justus-Liebig-University of Giessen, Institute for Insect Biotechnology, Heinrich-Buff-Ring 26-32, 35392 Giessen, Germany.
Andreas
A
Vilcinskas
D000818:Animals; D016000:Cluster Analysis; D020451:Contig Mapping; D016208:Databases, Factual; D003646:Debridement / Q000379:methods; D004175:Diptera / Q000201:enzymology*; D004798:Enzymes / Q000235:genetics / Q000302:isolation & purification*; D020869:Gene Expression Profiling; D019476:Insect Proteins / Q000235:genetics / Q000302:isolation & purification*; D007422:Intestines / Q000201:enzymology; D007814:Larva / Q000201:enzymology; D010447:Peptide Hydrolases / Q000737:chemistry; D059748:Proteolysis; D011550:Pseudomonas aeruginosa; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012469:Salivary Glands / Q000201:enzymology; D017423:Sequence Analysis, RNA; D013211:Staphylococcus aureus; D059467:Transcriptome; D014945:Wound Healing
D004798:Enzymes; D019476:Insect Proteins; D010447:Peptide Hydrolases