Franta Zdeněk, Vogel Heiko, Lehmann Rüdiger, Rupp Oliver, Goesmann Alexander, Vilcinskas Andreas
Department of Bioresources, Fraunhofer Institute for Molecular Biology and Applied Ecology, Winchesterstraße 2, 35394 Giessen, Germany.
Department of Entomology, Max Planck Institute for Chemical Ecology, Hans-Knöll-Straße 8, 07745 Jena, Germany.
Biomed Res Int. 2016;2016:8285428. doi: 10.1155/2016/8285428. Epub 2016 Mar 28.
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.
丝光绿蝇幼虫被用作顽固性和慢性伤口的替代治疗方法。它们的排泄物/分泌物含有促进组织清创、消毒或加速伤口愈合的分子,因此被认为是新型治疗化合物的潜在来源。在排泄物/分泌物中存在的各种物质中,已检测到多种促进伤口愈合过程的肽酶活性,但负责这些活性的肽酶大多仍未确定。为了探索这些酶,我们应用下一代测序技术分析了与排泄物/分泌物产生和/或消化相关的不同蛆组织(唾液腺、肠道和嗉囊)以及幼虫身体其他部分的转录组。结果,我们获得了超过1.238亿对末端读数,使用Trinity和Oases组装器进行从头组装,产生了41421个重叠群,N50重叠群长度为2.22kb,总长度为67.79Mb。针对MEROPS数据库的BLASTp分析在577个簇中鉴定出1729个重叠群,这些簇编码五种肽酶类别(丝氨酸、半胱氨酸、天冬氨酸、苏氨酸和金属肽酶),它们被分配到26个家族、48个家族和185个肽酶种类。这些个体酶在蛆组织中差异表达,包括与蛆排泄物/分泌物治疗效果相关的肽酶活性。