Lucifensin, Chymotrypsin Proteins and Molecular Characterization of Lucilia sericata


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Erdoğan E. , Karaca S.

Mikrobiyoloji Bulteni, vol.55, no.1, pp.81-90, 2021 (Journal Indexed in SCI)

  • Publication Type: Article / Letter
  • Volume: 55 Issue: 1
  • Publication Date: 2021
  • Doi Number: 10.5578/mb.20175
  • Title of Journal : Mikrobiyoloji Bulteni
  • Page Numbers: pp.81-90

Abstract

Lucilia sericata, one of the most common species of the Calliphoridae family, is found in large numbers

around droppings, garbage and carcasses. This fly species is important in medicine, forensics and veterinary

medicine. The larvae of the parasite are important both in veterinary medicine and in combating

of the animal diseases, as they cause significant losses in animal production. Since they are one of the

first fly colonies to settle on corpses, they can also be used in determining the time of death in the field

of forensic medicine. L.sericata larvae used in Maggot debridement treatment (MDT) which is a treatment

method with fly larvae, help wound healing by destroying necrotic tissues and infectious agents in

wounds. While the larvae protect themselves from polymicrobial flora with the proteins they secrete; at

the same time, they make an interesting contribution to wound healing with these molecules secreted.

One of the most important molecules discovered in recent years is lucimycin which has an antifungal

effect. In addition, lucifensin and chymotrypsin secretions have gained importance in recent years due

to their antibacterial effects and especially their effects on resistant gram-negative and positive bacteria.

There is a need for the discovery of the molecules that can be alternative in the treatment of non-healing

wounds or that can be applied together with existing antibiotics. It is necessary to investigate the antimicrobial

characterization of the compounds involved in maggot therapy and their mechanisms. The aim

of this study was to clone, molecular characterization and analysis of the antigenic structures of lucifensin

and chymotrypsin genes, which are important defensin molecules secreted by L.sericata larvae used in

MDT. Primarily, the cultivation of L.sericata colonies to be used in molecular studies were performed.

Later, RNA isolation and cDNA synthesis from larvae were carried out. Lucifensin and chymotrypsin

genes were individually inserted into the pJet1.2 plasmid by cloning reactions. The presence of the recombinant

plasmid was confirmed by PCR screening and DNA sequence analysis methods in all steps.

Nucleotide and amino acid based molecular characterizations of these two genes, which are important

larval components in wound treatment, have been made. Antigenic regions and three-dimensional structures

of the proteins were obtained. The isolate numbered MT495795 of the L.sericata lucifensin gene

and the isolate numbered MT495794 of the chymotrypsin gene were registered to GenBank. This data

reported for the first time in the Republic of Turkey will contribute to the literature. From the beginning

of the 20th century until the discovery of the antibiotics, MDT was applied especially on soldiers but did

not find much application area after the discovery of the antibiotics. Drug resistance, which is the most

important problem encountered in the treatment of the wounds today, has led to the recall of MDT and

its mechanism of action. In this study the data, obtained will constitute a source for the multidisciplinary

studies of the scientists from different fields on the discovery and applicability of the important molecules

in the treatment of the wounds.