Although the grasshopper Oxya chinensis sinuosa has long been used as a food in Korea, there is little data on itsfunctional effects. Thus we prepared and analyzed total RNA from the whole body of adult Escherichia coli non-immunizedand immunized Oxya chinensis sinuosa strains. Using an Illumina Hiseq sequencer, we generated a total of 66,555 pooledtranscriptome and singletons with and without Escherichia coli immunization, respectively. Then, we performed in silicoanalysis of the Oxya chinensis sinuosa transcriptome, using bioinformatics tools for screening putative antimicrobial peptides(AMPs) and 38 AMPs were finally selected and tested their antimicrobial activity of Gram positive, Gram negative bacteriaand antifungal activity with radial diffusion assay. As a result, 5 out of 38 AMPs showed the highest antimicrobial activityand antifungal activity against microbes and it also evidenced with no hemolytic activity.

Previously, we performed de novo RNA sequencing of Scolopendra subpinipes mutilans using high-throughput sequencing technology and identified several AMP candidates. Among them, a synthetic peptide (CP112) was designed based on the physicochemical properties of antimicrobial peptide such as length, charge, isoelectric point. Here, we have assessed the antimicrobial activities of CP112 against various microbes and the antioxidative effects. The results showed that CP112 had antimicrobial activities in radial diffusion assay and colony count assay. In addition, we found that CP112 bound to the surface of microorganisms via a specific interaction with lipoteichoic acid, lipopolysaccharide and peptidoglycan, which is one of bacteria cell wall components. Furthermore, CP112 has shown significant DPPH radical scavenging activity. Taken together, the results would be provided the basis for developing of peptide antibiotics and antioxidants.

Previously, we have performed de novo RNA sequencing of Scolpendra subpinipes mutilans using next generation sequencing technology and identified several AMP candidates. Among them, a synthetic peptide (scolopendrasin I) was designed based on SVM algorithm. In this study, we reported that the synthetic peptide scolopendrasin I had an antimicrobial and anticancer activity. As a result, scolopendrasin I showed antibacterial activities against Gram positive and Gram negative bacteria strains in radial diffusion assay and colony count assay without hemolytic activity. In addition, we confirmed that scolopendrasin I bound to the surface of bacteria via a specific interaction with lipoteichoic acid and lipopolysaccharide, which is one of bacteria cell membrane components. In addition, we found that scolopendrasin I had anticancer activities in the human leukemic T lymphocyte cell line Jurkat using MTS assay. In conclusion, our results suggested that scolopendrasin I could be useful for developing peptide antibiotics and anticancer agents.

To identify genes that are differentially expressed, we compared the mRNA expression profile of Harmonia axyridis larvae untreated and treated with LPS. We extracted mRNAs from the larvae with or without LPS treatment, and subjected them to ACP RT-PCR analysis using a combination of 120 arbitrary primers (ACP1-ACP120)and oligo (dT) primer (dT-ACP2). After synthesized cloning DNA from 37 DEGs, it practiced the sequencing homology analysis using BLAST search. Among the 37 DEGs differentially expressed, we identified a cDNA showing homology with previously reported antimicrobial peptide. A cDNA encoding a 82-mer propeptide was identified and its predicted molecular mass and pI was 9.25 kDa and 7.54, respectively. A 35-mer mature peptide was also selected and named herein as Hamoniasin. The antimicrobial activity of chemically synthesized peptide (Mou def 1~8) against human bacterial pathogens was investigate. the result showed all bacteria strains were susceptible to Mou def 2,8 with MIC values in the 32 uM range. And biological changes of the respective cells according to peptide (Mou def 8) treatment were compared. MTT assay was tested that treatment of Mou def 8 decreased cell viability in AML-2, Jurkat, U937 (maximum 200ug/ml, 24hours). That is, fragmentation of DNA, typical characteristics observed in the process of apoptosis, was confirmed in the nucleus of cells dying owing to Mou def 8 treatment.

Our previous study demonstrated that Coprisin, a peptide from Copris tripartitus infected with bacterial pathogens, has an antibacterial activity. We assessed in this study whether Coprisin caused cellular toxicity in various mammalian cell lines. Coprisin selectively caused a marked drop of cell viability in Jurkat T cells, U937 cells and AML-2 cells belonging to the human leukemia cells but not in Caki cells and Hela cells. Fragmentation of DNA, a maker of apoptosis, was also confirmed in theleukemia cell lines but not in other cells. The Coprisin-induced apoptosis in leukemia cells was mediated by AIF (apoptosis inducing factor), a caspase -independent pathway.

To examine the expression profile of oxidative stress responsive genes in Spodoptera litura, we constructed a cDNA library from S. litura injected with hydrogen peroxide (H2O2). Using a microarray chip composed of 2,964 cDNAs, we screened gene expression at 1, 3, 5, 7, and 9 h post H2O2 injection. Data were clustered into 15 groups of genes that behave similarly across each time course. Seventy-three genes were identified as being at least 2-fold up- or downregulated after treatment with H2O2 in S. litura. We constructed expressed sequence tags (ESTs) for genes that changed at least 2-fold after treatment with H2O2. The functional classification of these ESTs based on Gene Ontology showed that the ESTs are rich in genes involved in oxidoreductase activity (5.7%), defense (14.3%), cellular process (22.9%), and development (17.1%).

Inflammatory bowel disease (IBD) is a group of chronic disorders of unknown etiology characterized by inflammation of the gastrointestinal tract. Recent data showed that the development of IBD is associated with the interplay of genetic, bacterial, and environmental factors and dysregulation of the intestinal immune system. We investigated how the gut cells were repaired after injury in Drosophila melanogaster. In this study we made D. melanogaster intestine damage model by oral feeding with variety IBD inducer such as pathogenic bacteria Serratia marcescens, Dextran Sulfate Sodium (DSS) and bleomycin, because its function is very similar with human, even though D. melanogaster has relatively simple organism. We repeated oral feeding with variety IBD inducer and got the survival rate and 50% lethal dose (LD50). After feeding with IBD inducer, we investigated the change of the intestinal stem cells, innate immune-related gene expression, and apoptosis in D. melanogaster gut. We examined the Delta, stem cell marker, staining image in the gut after feeding with DSS and S. marcescens with LD50 concentration. The Delta positive cells greatly increased in gut cells damaged by DSS or S. marcescens. This result supports the idea that intestinal gut stem cells are increased after gut cell damage and play very important role in damaged cell repair. Expression level of antimicrobial peptides was dramatically up-regulation after gut damage. As a result of TUNEL (terminal deoxynucleotidyl transferase mediated X-dUTP nick end labeling) assay, we confirmed that cell death by apoptosis was very increased in DSS feeding flies. Accordingly, we suggest that D. melanogaster is a proper IBD model organism to study how intestine damage can be repaired.

A novel beetle antimicrobial protein from stimulated Copris tripartitus and the corresponding gene were isolated in parallel through differential display-PCR and expression in Escherichia coli. To find cDNA clones responsible for bacteria resistance, the suppression subtractive hybridization and GeneFishing differentially expressed genes system were employed in the dung beetle, Copris tripartitus immunized with lipopolysaccaride. One cDNA clone from eight subtracted clones was selected through dot blot analysis and confirmed by northern blot analysis. The 516-bp, selected cDNA clone was determined by 5' and 3'rapid amplication of cDNA ends and cloned into the GST fusion expression vector pGEX-4T-1 for expression of the protein. The expressed protein was predicted 14.7 kDa and inhibited the growth of gram-negative bacteria such as Escherichia coli and Pseudomonas aeruginosa. These results implied that the expressed protein is related to immune defense mechanism against microorganism.

식용 귀뚜라미를 고압증기멸균한 뒤 동결건조하였으며, 이를 분쇄하여 AL/PET/LLDPE film에 포장 후 25, 35 및 40℃에서 6개월 동안 저장하며 한 달 간격으로 품질변화를 측정하였다. 수분함량은 저장기간 중 모든 구에서 증가하 였으나, 저장 6개월 차에 저장온도별 유의적 차이는 없었다. 산가의 초기값은 1.83 mg KOH/g 이였으며, 저장 6개월 차에 25, 35 및 40℃ 저장구는 1.93, 2.04 및 2.15 mg KOH/g 으로 증가하였다. 지방산은 palmitic acid, oleic acid 및 linoleic acid가 24.79, 28.58 및 28.55 g/100 g으로 높은 함량 을 나타내었으며, palmitic acid 및 linoleic acid는 저장온도 별 증·감소의 경향이 뚜렷하지 않았으나, oleic acid는 저장 기간 중 감소하는 경향을 나타내었다. 일반세균 및 대장균 군은 저장기간 동안 검출되지 않았다. 관능검사의 경우 25 및 35℃ 저장구는 유의적 차이가 없으나, 40℃ 저장구는 유의적 차이를 나타내었다(p<0.05). 저장기간 중 품질에 영 향을 미치는 인자는 수분함량, 산가, oleic acid 및 flavor로 판단되었으며, 이들의 한계 기준을 각각 5%, 3 mg KOH/g, 26.5 g/100 g 및 6으로 설정하여 유통온도 25℃를 기준으로 VSLSF 프로그램으로 유통기한을 산출하였다. 수분함량, 산가, oleic acid 및 flavor의 유통기한을 산출한 결과 23, 61, 150 및 155개월이었으며, 유통기한이 가장 짧은 항목에 안전계수 0.8을 적용하여 18개월로 설정하였다. 이상의 결 과는 제조 후 25℃ 유통조건에서 18개월까지는 귀뚜라미 분말의 품질 안정성이 유지되는 것으로 판단되었다.

Insects have gained increasing attention as an alternative protein and nutrient rich food source for humans. This study was conducted to investigate the physicochemical characteristics and harmful components of edible crickets (Gryllus bimaculatus) in the 6 districts of Yeonggwang (YG), Jeongseon (JS), Wonju (WJ), Hwaseong (HS), Geochang (GC), and Chungju (CJ). The average crude protein and crude lipid contents on a dry basis were 64.34% and 16.60%, respectively. The crude protein content of CJ was the highest (67.40%), whereas YG (59.42%) had the lowest content. On the other hand, the crude fat content of YG was the highest (20.61%), whereas CJ (14.04%) had the lowest content. The unsaturated fatty acid contents were 57.97-63.93 g/100 g of the total fatty acid content in the crickets of the 6 districts. The major fatty acids of the crickets in the 6 districts were palmitic acid, oleic acid, and linoleic acid. Among the essential amino acids, valine, leucine, and lysine were the most abundant. GC had the highest total amino acids (57.93 g/100 g), whereas YG (48.65 g/100 g) had the lowest. Major mineral contents included potassium (K, 0.92~1.01 mg/100 mg) and phosphorus (P, 0.74~0.88 mg/100 mg). The mineral composition was fairly similar among the crickets. Crickets in the 6 districts were verified to have safe levels of residual heavy metals according to the Korea Food & Drug Administration (KFDA) advisory levels.

본 연구는 귀뚜라미 소재를 식품으로서의 활용도를 높이기 위하여 다양한 가공방법에 따른 품질 변화를 조사하였다. 일반성분 중 조단백과 조회분 함량은 동결건조 처리구에서 가장 높게 나타났고, 수분함량과 조지방은 튀김건조 처리구에서 가장 높게 나타났으며, 조섬유는 각각의 처리구에서 유사하게 나타났다. 열풍건조 처리구의 키틴함량은 약 12%로 튀김건조 처리구의 6%의 약 2배 높은 함량을 나타내었다. 동결건조 귀뚜라미 시료의 지방 100 g 중에서 불포화지방산이 63.55 g으로 가장 높게 측정되었고, 포화지방산은 튀김건조 처리구가 지방 100 g 중에서 31.88 g으로 가장 높은 함량을 나타내었다. 무기질 함량은 동결건조 처리구에서 가장 높게 나타났으며, 열풍건조, 볶음건조 및 튀김건조 순으로 나타났다. 건조조건에 따른 귀뚜라미의 안전성을 알아보기 위하여 중금속과 미생물 분석을 한 결과, 카드뮴과 수은이 검출되었으나 이는 각각 중금속의 식약처 고시 기준인 0.2 ppm, 0.5 ppm 보다 낮은 수준으로 나타났으며, 납과 비소는 모든 처리구에서 검출되지 않았다. 또한 병원성 미생물도 모든 처리구에서 검출되지 않았다