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Kyungjae Andrew Yoon,Kyungmun Kim,A-Young Kim,Young Han Park,Woo Young Bang,Chang Hwan Bae,Joo-Hong Yeo,Young Ho Koh,Si Hyeock Lee 한국응용곤충학회 2016 한국응용곤충학회 학술대회논문집 Vol.2016 No.10
Bombolitin is a venom peptide originally isolated from bumblebees and possesses various biological activities, including hemolytic activity. Bombolitins exhibit amphipathic α-helical structure in lipid-membrane-mimicking environments. To investigate their pharmacological and toxicological properties, anti-tumor, anti-microbial and cytotoxic activities of bombolitins from Bombus ardens and Bombus ussurensis were evaluated. Bombolitins of the two species exhibited extremely high anti-tumor activity against ovarian tumor cells SK-OV-3 and NIH; OVCAR-3 at 25-50 μM, which is 2-fold more potent than other wasp venom peptides studied to date (Yoon et al., 2015; Yoon et al., 2016). The two bombolitins showed significantly high antimicrobial activity. However, bombolotin of B. ussurensis showed no antimicrobial activity against Escherichia coli. In addition to their high levels of anti-tumor activity, bombolitins showed considerable levels of hemolytic activity. Thus, to utilize bombolitins as a potential candidate for anti-tumor peptide drugs, further studies for reducing cytotoxic properties of bombolitns is essential.
Specific anti-tumor activities in venom peptides of lesser paper wasp Parapolybia varia
Kyungjae Andrew Yoon,Kyungmun Kim,A-Young Kim,Young Han Park,Woo Young Bang,Chang Mu Kim,Young Ho Koh,Si Hyeock Lee 한국응용곤충학회 2016 한국응용곤충학회 학술대회논문집 Vol.2016 No.04
The lesser paper wasp, Parapolybia varia, belongs to large subfamily Polistinae and is distributed in Middle East, the Indo-Papuan region and East Asia. P. varia is known to become aggressive when disturbed for defending their colonies, resulting in fatal envenomation. Vespid chemotactic peptide (VCP) and vespakinin have recently been determined to be the top two genes most abundantly transcribed in venom glands of P. varia. To investigate the pharmacological and toxicological properties of VCP and vespakinin, their antitumor, antimicrobial, and cytotoxic activities were evaluated. VCP exhibited a significantly high antitumor activity against ovarian tumor cell SK-OV-3 at 100 M. VCP also showed higher hemolytic activity than vespakinin. Antimicrobial activity was only observed with VCP against yeast Candida albicans at 1 mM. Since VCP showed a relatively low hemolytic activity but a considerable level of antitumor activity, it has further merits to be exploited as a potential antitumor agent with reduced side effects on normal cells.
Comparison of Endogenous Cellulase Genes from Four Termite Species with Different Habitats
Kyungjae Andrew Yoon,Young Ho Kim,Si Hyeock Lee 한국응용곤충학회 2013 한국응용곤충학회 학술대회논문집 Vol.2013 No.04
To search for a variety of cellulase genes from termites with different habitats consuming different foods, we collected three species (Neotermes spp., Odontotermes spp., Macrotermes spp.) from the wood and one species (Nasutitermes spp.) from the cow dung. Total RNA was isolated both from alimentary track tissues containing paunch and from other tissues, and used for the suppression subtractive hybridization (SSH). The resulting EST libraries were sequenced and searched by BLAST to identify cellulase genes. A total of 16 cellulase genes were found from the wood-dwelling termites whereas 4 cellulase genes from the cow dung-dwelling termites. Endo-beta-1,4-glucanase and beta-glucosidase were identified as the most abundant cellulase from the wood-dwelling termites and cow dung-dwelling termites, respectively. This finding suggests that cellulase profiles are significantly different depending on the termite’s habitat and food. In addition, we analyzed phylogenetic relationships among the cellulase genes along with other cellulase genes reported to date. All cellulase genes appeared to be originated from endosymbioants without any hint of horizontal gene transfer. Functional expression of endo-beta-1,4-glucanase using a baculovirus expression system is in progress to characterize its enzymatic properties.
Kyungjae Andrew Yoon,Young Ho Kim,Si Hyeock Lee 한국응용곤충학회 2013 한국응용곤충학회 학술대회논문집 Vol.2013 No.10
The aim of this study is to analyze the functional activity of an endo-β-1, 4-glucanase from the wood dwelling lower termite Coptotermes gestroi. Full length cDNA sequences of the endo-β-1,4-glucanase were obtained by primer walking in conjunction with Rapid Amplification cDNA Ends. With the obtained full length sequences, primers for amplifying open reading frame (ORF) excluding the signal peptide and glycophosphatidylinositol anchor were designed. Amplified endo-β-1,4-glucanase fragment was cloned and expressed using pET30(+) expression vector in BL21 E.coli strain. Expression of endo-β-1,4-glucanase was confirmed by Western blotting and the result revealed that only full ORF was expressed. The cellulase activity of protein preparations from the induced and non-induced cells was analyzed with Congo Red assay with the cellulase from Aspergillus niger (Sigma Aldrich) as a positive control. The activity of C. gestroi endo-β-1,4-glucanase was significantly higher than those observed in the positive control and the enzyme preparation from non-induced cells. Therefore, this study confirmed that C. gestroi endo-β-1,4-glucanase had a function of cellulose hydrolysis.
Comparative transcriptomic analysis of social hornets Vespa crabro and Vespa analis
Kyungjae Andrew Yoon,Kyungmun Kim,Hyo-min Ah,Ki-Gyoung Kim,Young Ho Koh,Young Ho Koh,Si Hyeock Lee 한국응용곤충학회 2015 한국응용곤충학회 학술대회논문집 Vol.2015 No.10
The hornets Vespa crabro and V. analis are widely distributed in Asia and are known to be aggressive when disturbed, resulting in frequent stinging accidents. To investigate the differences in venom properties and toxicities between these two hornets, the transcriptomic profiles of venom glands, in conjunction with the venom components, were analyzed and compared. A total of 35 venom-specific genes were identified in both venom gland transcriptomes, but their transcriptional profiles were different between V. crabro and V. analis. In addition, the major venom components were identified and confirmed by mass spectroscopy. Prepromastoparan, vespid chemotactic precursor and vespakinin were the top three genes most prevalently transcribed in the venom gland of V. crabro, and their transcription rates were 112-, 16- and 161-fold higher, respectively, compared with those in V. analis, as judged by FPKM values. In the venom gland of V. analis, however, vespid chemotactic precursor was the most abundantly transcribed gene, followed by premastoparan and vespakinin. In general, most major venom genes were more abundantly expressed in V. crabro, whereas some minor venom genes exhibited higher transcription rates in V. analis, including muscle LIM protein, troponin, paramyosin, calponin, etc. Our findings reveal that the overall venom components of V. crabro and V. analis are similar, but that their expression profiles and levels are considerably different. The comparison of venom gland transcriptomes suggests that V. crabro likely produces venom with more highly enriched major venom components, which has potentially higher toxicity compared with V. analis venom.
Kyungjae Andrew Yoon,Young Han Park,Young Ho Koh,Si Hyeock Lee 한국응용곤충학회 2017 한국응용곤충학회 학술대회논문집 Vol.2017 No.10
Biological activities of bombolitins from Bombus ardens, B. consobrinus, B. terrestris and B. ussurensis (bombolitinsA, C, T and U, respectively) were examined using hemolytic, anti-microbial, anti-fungal and anti-tumor activity assays.Among the four bombolitins tested, bombolitin T showed the highest hemolytic and anti-tumor activities. All bombolitinsexhibited strong anti-microbial and anti-fungal activities, and bombolitin A specifically possessed the highest anti-microbialactivity against the Gram-negative bacteria Escherichia coli. Circular dichroism spectrometry analysis revealed that allfour bombolitins had over 61.7% and 45.5% of α-helicity in 30 mM sodium dodecyl sulfate and 50% trifluoroethanolbuffers, respectively, which form lipid-membrane-mimicking environments. Bombolitin T showed the lowest IC50 valuesof 8.5 μM and 8.8 μM against SK-OV-3 and NIH-OVCAR-3 cell lines, respectively, after 72 h of treatment, but itsrelative hemolytic activity at a concentration of 200 μM was 2.3-fold higher than that of 0.1% Triton X-100. Thisstudy provides new information on the biological and molecular properties of venom peptides of bumblebees. However,further studies on reducing cytotoxicity of bombolitins are needed for designing selective anti-tumor peptides.
Detection of the Bacterial Symbiosis in the Pea Aphid Acrythosiphon pisum.
Kyungjae Andrew Yoon,Young Ho Kim,Si Hyeock Lee 한국응용곤충학회 2012 한국응용곤충학회 학술대회논문집 Vol.2012 No.10
Many previous ecological studies on three major bacterial symbionts and a newly discovered symbiont PAXS (pea aphid X-type symbiont) in the pea aphid Acrythosiphon pisum have shown that these symbionts are associated with the expression of a variety of host phenotypes, including resistance to parasitoid Aphidius ervi and tolerance to heat stress. The principal role of all four symbionts “Candidatus Serratia symbiotica”, “Candidatus Hamiltonella defensa”, “Candidatus Regiella insecticola”, and PAXS is to protect aphids against abiotic stress by preserving the cells in which most of symbionts dwell and by reducing the rate of parasitism. In this experiment, we detected endosymbionts from four aphid clones by means of genomic DNA extraction, PCR with gene specific primer, and restriction enzyme cutting. The patterns of PCR and restriction enzyme cutting were all identical in the four aphid clones. In order to specifically identify the endosymbiont, we searched the sequences using BLAST. The BLAST search revealed that nucleotide sequences of the symbiont were 98% identical to Serratia entomophil. S. entomophil is also known to provide tolerance to heat stress, resistance to parasitoid wasps, and restoration of reproduction in aphids, suggesting its role in host protection.
Kyungjae Andrew Yoon,Do Eun Lee,Woo Young Bang,Chang Hwan Bae,Joo-Hong Yeo,Young Ho Koh,Si Hyeock Lee 한국응용곤충학회 2018 한국응용곤충학회 학술대회논문집 Vol.2018 No.10
To identify the venom components and their expression patterns of some Aculeata bees/wasps, venom gland-specific transcriptome analysis was conducted. FPKM values were normalized with the average of the transcription level of reference gene (a-tubulin). Common components in both solitary and social wasp venoms include hyaluronidase, phospholipase A2, metalloendopeptidase, etc. Although it has been expected that more diverse bioactive components with the functions of prey inactivation and physiology manipulation are present in solitary wasps, the information on venom compositions of solitary wasps obtained in this study was not sufficient to generalizae this notion. Nevertheless, some neurotoxic peptides (e.g., pompilidotoxin and dendrotoxin-like peptide) and proteins (e.g., insuline-like peptide binding protein) appear to be specific to solitary wasp venom. In contrast, several proteins, such as venom allergen 5 protein, venom acid phosphatase, and various phospholipases, appear to be relatively more abundant in social wasp venom. In the venom gland trancsriptome of bumblebees, major allergens or pain producing factors were barely identified, implying that bumblebee venoms are relatively less toxic than those of social or solitary wasps.