Utilizing Red Spotted Apollo Butterfly Transcriptome to Identify Antimicrobial Peptide Candidates against Porphyromonas gingivalis

Abstract

Classical antibiotics are the foremost treatment strategy against microbial infections. Overuse of this has led to the evolution of antimicrobial resistance. Antimicrobial peptides (AMPs) are natural defense elements present across many species including humans, insects, bacteria, and plants. Insect AMPs are our area of interest, because of their stronger abilities in host defense. We have deciphered AMPs from an endangered species Parnassius bremeri, commonly known as the red spotted apollo butterfly. It belongs to the second largest insect order Lepidoptera, comprised of butterflies and moths, and lives in the high altitudes of Russia, China, and Korea. We aimed at identifying the AMPs from the larvae stages. The rationale of choosing this stage is that the P. bremeri larvae development occurs at extremely low temperature conditions, which might serve as external stimuli for AMP production. RNA was isolated from larvae (L1 to L5) instar stages and subjected to next generation sequencing. The transcriptomes obtained were curated in in-silico pipelines. The peptides obtained were screened for requisite AMP physicochemical properties and in vitro antimicrobial activity. With the sequential screening and validation, we obtained fifteen candidate AMPs. One peptide TPS–032 showed promising antimicrobial activity against Porphyromonas gingivalis, a primary causative organism of periodontitis.

Developmental Transcriptome Analysis of Red-Spotted Apollo Butterfly, Parnassius bremeri

“…Few researchers have focused on developing molecular markers, such as SSRs [ 22 ]. Our previous transcriptome study dealt with the identification of antimicrobial peptide candidates against the primary periodontitis pathogen Porphyromonas gingivalis [ 16 ]. In general, the genus Parnassius is an altitudinal butterfly, and they are thought to appear in different colors depending on altitude.…”

“…The genome of the nearest species of P. bremeri, P. apollo, Papilio xutus, Papilio machaon and P. bianor have been studied recently [ 13 , 14 , 15 ]. In addition, in recent times we have identified antimicrobial peptide candidates in P. bremeri against the causative agent of periodontitis, Porphyromonas gingivalis [ 16 ]. However, the mechanisms involved in the development of P. bremeri from egg stage to adult stage remain unclear.…”

Parnassin, a Novel Therapeutic Peptide, Alleviates Skin Lesions in a DNCB-Induced Atopic Dermatitis Mouse Model

“…Parnassin is a novel therapeutic peptide deciphered from P. bremeri transcriptome data using our in silico curation pipeline [20]. Briefly, total RNA was isolated from larva, sequenced and de novo assembled.…”

“…Parnassin was synthesized and purified at Lugen Sci Co., Ltd. (Bucheon, Republic of Korea) according to the methods described previously [20]. Parnassin was dissolved in distilled phosphate buffered saline (PBS) at a concentration of 1 mg/mL.…”

“…Parnassin (KTFR) is a tetrapeptide with anti-bacterial activity curated from Parnassius bremeri (P. bremeri, red-spotted Apollo butterfly) transcriptome data using our in silico therapeutic peptide screening platform [20]. Therapeutic peptides with the anti-microbial property have also been shown to possess immunomodulatory effects and wound healing efficacy [21,22].…”