Biochemical and Functional Characterization of Glycoside Hydrolase Family 16 Genes in Aedes aegypti Larvae: Identification of the Major Digestive β-1,3-Glucanase

Abstract

Insect β-1,3-glucanases belong to Glycoside Hydrolase Family 16 (GHF16) and are involved in digestion of detritus and plant hemicellulose. In this work, we investigated the role of GHF16 genes in Aedes aegypti larvae, due to their detritivore diet. Aedes aegypti genome has six genes belonging to GHF16 (Aae GH16.1 – Aae GH16.6), containing two to six exons. Sequence analysis suggests that five of these GHF16 sequences (Aae GH16.1, 2, 3, 5, and 6) contain the conserved catalytic residues of this family and correspond to glucanases. All genomes of Nematocera analyzed showed putative gene duplications corresponding to these sequences. Aae GH16.4 has no conserved catalytic residues and is probably a β-1,3-glucan binding protein involved in the activation of innate immune responses. Additionally, Ae. aegypti larvae contain significant β-1,3-glucanase activities in the head, gut and rest of body. These activities have optimum pH about 5–6 and molecular masses between 41 and 150 kDa. All GHF16 genes above showed different levels of expression in the larval head, gut or rest of the body. Knock-down of AeGH16.5 resulted in survival and pupation rates lower than controls (dsGFP and water treated). However, under stress conditions, severe mortalities were observed in AeGH16.1 and AeGH16.6 knocked-down larvae. Enzymatic assays of β-1,3-glucanase in AeGH16.5 silenced larvae exhibited lower activity in the gut and no change in the rest of the body. Chromatographic activity profiles from gut samples after GH16.5 silencing showed suppression of enzymatic activity, suggesting that this gene codes for the digestive larval β-1,3-glucanase of Ae. aegypti . This gene and enzyme are attractive targets for new control strategies, based on the impairment of normal gut physiology.

Myriapod genomes reveal ancestral horizontal gene transfer and hormonal gene loss in millipedes

“…Various types of glucoside hydrolases are present in the genomes of phytophagous insects including beetles and moths 26 – 28 . While in the nematode Aphelenchoides besseyi and Aedes aegypti mosquito larvae, GH16 is suggested to be associated with glucan rich diets 29 , 30 . Previous studies on centipedes have also illustrated the presence of GH in venoms, where they are postulated to act as “spreading factors” that promote the pathological effects of other venom elements after envenomation, and also as antimicrobial factors 31 .…”

Rhodnius prolixus Hemolymph Immuno-Physiology: Deciphering the Systemic Immune Response Triggered by Trypanosoma cruzi Establishment in the Vector Using Quantitative Proteomics

“…In addition to their immune role, these proteins seem to have a digestive implication when they are expressed in the insect's midgut [42]. Interestingly, we have shown in the comparative proteome of R. prolixus midgut [43,44] that T1HU92 (ML domain protein) expression level increases in response to blood feeding.…”

Knockdown of Anopheles dirus far upstream element‐binding protein gene lower oocyst numbers of Plasmodium vivax

“…Proteins containing ABHD, including kraken protein, have diverse catalytic functions such as proteases, esterases, nucleases, glycosidases, and phosphatases. Most studies focusing on the hydrolase functions of mosquitoes, involving glucanases (Souza et al, 2019), epoxide hydrolases (Xu et al, 2014), and acetylcholinesterases (Dou et al, 2013), have suggested that they were not related to the response to pathogen infection. However, the range of oocyst numbers was variable depending on the blood sample used, T A B L E 5 The total binding energy of 20 top-scoring compounds which bound to AdFUEBP homology model.…”