호박벌의 휴면 제어 관련 유전자 탐색 및 특성 분석

황재삼 ( Jae Sam Hwang ),박인균 ( In Gyun Park ),윤은영 ( Eun Young Yun ),정덕웅 ( Duck Oung Jung ),윤형주 ( Hyung Joo Yoon ),이상범 ( Sang Beom Lee ),김성렬 ( Seong Ryul Kim ),이준하 ( Joon Ha Lee ),김인우 ( In Woo Kim ),권용남 ( 한국잠사학회 2012 한국잠사곤충학회지 Vol.50 No.2

To find diapause-related genes, we were performed by differential hybridization with three types of [α-32P]dCTP-labeled total cDNA probes synthesized from diapause-prepared, diapause-maintained and diapause-activated stage of Bombus ignitus queen. Nine individual cDNA clones were found to be differentially expressed in diapause-maintained and diapause- activated stage. Among these clones, BIDC9(BIDC ; Bombus ignitus differentially expressed clone) was analyzed through full-length sequencing and expression pattern analysis. This clone was specifically expressed in the thorax organ. The effect of Juvenile hormone analog(JHA) and CO2 treatment was examined. JHA treatment induced the expression of BIDC9 cDNA clone abruptly after 4 day of treatment. CO2 treatment induced also the clone after 2 day of treatment. BIDC9 cDNA was identified as Bombus ignitus diapause gene contained an open reading frame of 1376 bp encoding 255 amino acids.

RAPD 마커를 이용한 멧누에와 집누에 계통간의 분자적 유연관계 분석

황재삼,이진성,구태원,강현아,손해룡,김호락,Hwang, Jae-Sam,Lee, Jin-Sung,Goo, Tae-Won,Kang, Hyun-Ah,Sohn, Hae-Ryong,Kim, Ho-Rak 한국생명과학회 1998 생명과학회지 Vol.8 No.4

본 연구는 RAPD마커를 이용, 멧누에와 집누에의 분자적 유연관계를 분석하였다. 공시한 35개의 primer에서 166개의 RAPD마커를 얻었으며, 이들 마커를 UPGMA에 의해 분석한 결과, 멧누에와 분자적 유사계수가 가장 낮은 품종은 잠305였고, 가장 높은 품종은 Bibaekjam이었다. 또한, 분자적 유사계수 0.55에서 멧누에와 집누에 계통군으로 분류되고, 0.60에서 3개의 아군 그룹과 2개의 독립개체로 분류되었다. 제1아군에는 J111(일본종계),$pnd^{ps}$(일본종계), Bibaekjam(일본종계)이, 제2아군에는 Galwon(중국종계), C18(중국종계), od yujam JAM306(중국종계), C108(중국종계)이, 제3아군에는 R-hwang(중국종계)이 포함되어 있었고, zebra(유럽종계)와 JAM305(일본종게)는 독립개체로 분류되었다. The molecular relationships have analyzed between the Bombyx mandarina(wild silkworm) and Bombyx mori strains (domesticated silkworm, geographical silkworms). A total of 166 polymorphic RAPD markers amplified from 35 different primers were used to analyze the molecular relationships among thirteen silkworm strains. The genetic similarity coefficient between Bombyx mandarina and Jam305 showed the lowest genetic similarity value with 0.451, Bombyx mandarina and Bibaekjam showed the highest genetic similarity value with 0.958. These strains were classified into Bombyx mandarina(a wild silkworm) and Bombyx mori(twelve domesticated silkworm) groups upon the genetic similary coefficient of 0.55. Further classificient of 0.60; the 1st sub-group (J111, Bibaekjam, $pnd^{ps}$), the 2nd sub-group (Galwon, C18, od yujam, JAM306, C108), the 3rd sub-group(R-hwang, p50), the 4th sub-group(zebra) and the 5th sub-group(JAM305). According to this study, RAPD markers seems to be a valuable tool for molecular relationships and classification among the silkworms.

견사곤충에서 Mariner Transposase-like Element의 분자적 동정

이진성,황재삼,김용성,서동상,Lee, Jin Sung,Hwang, Jae Sam,Kim, Yong Sung,Suh, DongSang 한국생명과학회 1998 생명과학회지 Vol.8 No.4

누에, 멧누에, 천잠 및 작잠에서의 형질전환용 vector 개발을 위한 첫단계로 이미 인간을 포함한 여러 곤충에서 그 존재가 확인된 mariner transposable element가 이들 견사 곤충에 존재하는지의 여부를 탐색하기 위해서 이미 보고된Drosophila mauritiana와 Hyalophora ceropia의 mariner transposase 유전자의 고도로 보존성이 높은 부위에 대한 degenerctive primer pair를 제작하여 이들의 존재 여부를 탐색하였다. 결과적으로 예상되는 약 500bp의 PCR product가 관찰됨에 따라서 모든 견사곤충의 genome에는 maminer-like element(MLE)가 존재한다는 것을 간접적으로 확인할 수 있었다. 이미 cloning된 pBmoMAR를 probe DNA로하여 southern hybridization을 수행한 결과에서도 확인 할 수 있었으며, 각 견사층의 genome내에 high copy로 존재한다는 것을 추정할 수 있었다. 따라서 이와 같은 결과는 견사곤충에서 MLE가 vector system으로써 개발될 수 있는 기초자료로 제공될 수 있을 것이다. As a first step for developing universal genetic transformation vector of dilkmoths., we identified the presence of mariner-like element(MLE) which is one of transposable element discovered from many insects to human species, from Bombyx mori, Bombyx mandarina, Antherae yammamai and Antherae pernyi. We used a degenerative primer pair designed from a transposase gene of Drosophila mauritiana and Hyalophora ceropia MLE. As results, major PCR product of 500bp expected as a part of transposase of MLE was detected from all the slkmoths used of this study using these primer. And hybridization assay using pBmoMAR as a probe DNA that was previously cloned from Bombyx mori by the same primer pair, confirmed the presence of MLE from all the silkmoths. This assay showed also that the endogenous MLE in genome of the silkworm is present as high copy number unlikely Drosophila mauritiana which has 10-20 copy number. This data will be a fundamental genetic information for developing mariner-derived vector to transform the silkmoths and other useful insects.

누에에서의 Mariner 유사 전이인자유전자의 동정

이진성,황재삼,김용성,서동상,권오유,Lee, Jin-Sung,Hwang, Jae-Sam,Kim, Yong-Sung,Suh, Dong-Sang,Kwon, O-Yu 한국생명과학회 1998 생명과학회지 Vol.8 No.3

We have cloned an internal fragment of the putative transoisase gene of MLE in the silkworm, Bombyx mori, using PCR method with degenerative oligonucleotide primers designed to represent regions of amino acids encoding transposase. The resulting PCR clone, designed as BmoMAR, cords a partial ORF(152 a.a.) of MLE in which interrupted by five stop codons, and the sequence of its deduced amino acids showed 37% homology with Mos1, an active mariner, from Drosophila mauritiana. Furthermore, the BmoMAR exhibits nucleotide and amino acid homology with 59% and 37% from Apis mellifera and D. mauritiana 7.9 clone, respectively. This result strongly that a MLE is present in the genome of B. mori. 이미 밝혀져 있는 mariner 전이인자의 전이효소를 암호화하는 부위에 대하여 퇴화성 primer를 사용하여 PCR 방법에 의해 누에(Bombyx mori)에서 ariner 유사 전이닌자의 잠정적인 전이효소 부위를 클로닝 하였다. BmoMAR로 망명된 이 PCR 클론으로부터 추론된 아미노산은 152개로 다섯 개의 종결코돈이 삽입되어 있었으며, Drosophila mauritiana의 active Mos 1에 37%의 아미노산 상동성을 보였다. 또한, 기존의 곤충들에서 밝혀진 mariner-like element에 대한 상동성은 DNA 수주에서는 Apis mellifera에 59% 그리고 아미노산 수준에서는 D. mauritiana 7.9 clone에 37% 상동성을 보였다. 이 결과는 mariner-like element가 B. mori에도 존재하고 있지만. 이들 전이인자의 전이효소를 암호화하는 부위에 종결코돈이 발견되는 것으로 보아서 비활성 전이인자 혹은 일존의 selenoprotein으로 추정된다.

새로운 식품원료로 등록된 식용곤충의 영양성분 비교 분석

백민희(Minhee Baek),황재삼(Jae-Sam Hwang),김미애(Mi Ae Kim),김수희(Soo-Hee Kim),구태원(Tae-Won Goo),윤은영(Eun-Young Yun) 한국생명과학회 2017 생명과학회지 Vol.27 No.3

본 연구에서는 갈색거저리, 흰점박이꽃무지 및 장수풍뎅이 유충의 영양성분을 비교분석하였고, 이를 통해 식용곤충 이용 시 곤충 선택 기준확립에 기초 자료로 활용될 수 있을 것으로 사료된다. 3종 식용곤충의 영양성분 비교결과, 조단백질, 조지방, 탄수화물은 각각 흰점박이꽃무지, 갈색거저리 및 장수풍뎅이 유충에서 가장 높았다. 아미노산 함량 비교 분석 결과, 총 아미노산 함량과 필수 아미노산의 함량은 갈색거저리 유충에서 가장 높았다. 지방산의 조성은 갈색거저리와 흰점박이꽃무지 유충이 유사하였으며, 이들은 장수풍뎅이 유충에 비해 불포화 지방산 함량이 높았다. 또한, 무기질 함량의 비교 분석 결과, 칼슘과 철은 장수풍뎅이 유충에서 가장 높은 반면 인과 칼륨은 흰점박이꽃무지 유충에 가장 많이 함유되어 있었다. 위와 같은 결과를 통해 각각의 곤충이 함유하는 영양성분에 따라 노인, 어린이 및 임산부 등 다양한 영양 요구를 가지는 대상에 따라 필요한 곤충을 선택하여 섭취가 가능할 것이며 식용곤충을 다양하게 조합하는 경우 보다 풍부한 영양분을 섭취할 수 있을 것으로 사료된다. In this study, the nutritional components of Tenebrio molitor, Protaetia brevitarsis, and Allomyrina dichotoma larvae, which have been registered as novel foods, were analyzed and compared to expand the diversity of selection criteria for edible insects. The contents of crude components, amino acids, fatty acids, and minerals were analyzed. According to the results of comparative analysis of edible insects, crude proteins were abundant in all three kinds of insects. The content of crude fat was the highest in T. molitor, and the content of carbohydrate was the highest in A. dichotoma. When comparing the composition of amino acids, total amino acid content and essential amino acids were the highest in T. molitor larvae. In T. molitor and P. brevitarsis larvae, the compositions of fatty acids were similar, with higher amounts of unsaturated fatty acids than in A. dichotoma. In terms of mineral content, A. dichotoma contained the highest amounts of calcium and iron, whereas P. brevitarsis contained the highest amounts of phosphorus and potassium. With these results, it is expected that edible insects could be selected according to nutritional demand. In addition, multiple combinations of edible insects will offer a richer intake of nutrients.

LPS로 염증 유도된 RAW 264.7세포에 대한 참콩풍뎅이(Popillia flavosellata) 에탄올 추출물의 항염증 효과

윤영일(Young-Il Yoon),황재삼(Jae-Sam Hwang),김미애(Mi-Ae Kim),안미영(Mi Young Ahn),이영보(Young-Bo Lee),한명세(Myung Sae Han),구태원(Tae-Won Goo),윤은영(Eun-Young Yun) 한국생명과학회 2015 생명과학회지 Vol.25 No.9

본 연구에서는 참콩풍뎅이(Popillia flavosellata) 에탄올 추출물(PFE)의 항염증 효능을 분석하기 위해 PFE를 농도별(500, 1,000, 2,000 ㎍/ml)로 대식세포인 RAW 264.7에 처리 시 최고 처리농도인 2,000 ㎍/ml까지 통계적인 유의성 있는 독성이 없음을 확인하였다. LPS (100 ng/ml)로 염증 유도된 RAW 264.7 세포에 PFE를 농도별(500, 1,000, 2,000 ㎍/ml)로 동시 처리 시 농도 의존적으로 염증성사이토카인인 TNF-α와 IL-6의 단백질 생성을 통계적인 유의성(p<0.001)있게 억제함을 확인하였다. 또한 염증 유도된 RAW 264.7 세포에 PFE 동시 처리 시 NF-κB p65의 핵으로 이동이 차단됨과 iNOS와 COX-2의 단백질 발현을 감소시키는 것을 확인하였다. 이상의 연구결과를 통해 참콩풍뎅이는 염증에 의해 활성화된 TLR-4 신호전달과정을 조절하는 NF-κB p65의 활성과 염증성사이토카인 TNF-α와 IL-6의 생성 및 염증성효소 iNOS와 COX-2의 생성을 억제하는 항염증 효능이 있음을 확인하였다. The beetle Popillia flavosellata has been no reported its functional effects. In this study, we investigated the anti-inflammatory effect of P. flavosellata ethanol extract (PFE) on RAW 264.7 mouse macrophage cells treated with lipopolysaccharide (LPS) for the induction of inflammation. First, we examined the cytotoxicity of PFE in the RAW 264.7 cells at a concentration of 2,000 μg/ml or less. To evaluate the anti-inflammatory effects of PFE, we investigated the expression levels of proinflammatory cytokines, such as tumor necrosis factor (TNF)-α and interleukin (IL)-6, and proinflammatory enzymes, such as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in LPS-induced RAW 264.7 cells. In addition, we examined whether PFE inhibited the translocation of nuclear factor kappa B (NF-κB) p65 into the nucleus in the LPS-induced RAW 264.7 cells. We found that the protein levels of TNF-α and IL-6 were decreased in the LPS-induced RAW 264.7 cells after the treatment with PFE in a dose-dependent manner. In addition, we confirmed that PFE inhibited the translocation of NF-κB p65 into the nucleus, as well as the protein expression levels of iNOS and COX-2. Accordingly, we propose that PFE exerts an anti-inflammatory effect through the down-regulation of NF-κB p65, TNF-α, IL-6, iNOS, and COX-2 via the toll like receptor (TLR)-4 inflammatory signaling pathway.

곤충식품 개발 현황 및 전망

윤은영(Eun-Young Yun),황재삼(Jae-Sam Hwang) 한국식품과학회 2016 식품과학과 산업 Vol.49 No.4

The reasons for selecting insects as future food are high growth and feed conversion rates, reproduce quickly, low environmental footprint, valuable source of nutrients, and source of a various undeveloped functional materials. Since 2014, Tenebrio molitor larva, Protaetia brevitarsis larva, Allomyrina dichotoma larva, and Gryllus bimaculatus were registered as new food in Korea because it has been scientifically proven that they are not harmful to eat and nutritious. Therefore they can be legally produced and sold as food. Accordingly, there are 7 species of edible insects including grasshopper, silkworm pupa, and Baekgangjam in Korea. To improve aversion to edible insects, using their powder, chop, and gravy hidden their morphology, we developed more than 100 different kinds of recipes for general food menu, held several times tasting events to be familiar with edible insect food, and published cookbooks. Moreover, we developed more than 50 kinds of recipes for patients. To eat the insect food more and more people, we have been analyzed various function of insects. If health food based on the results of functional analysis is developed, edible insect’s value will be raised. If various insect foods are developed and consumed by people, edible insect market will grow up more than 100 billion won in 2020.

국산 및 중국산 갈색거저리(Tenebrio molitor)의 영양성분 및 유해물질 비교분석

유정미(Jeongmi Yoo),황재삼(Jae-Sam Hwang),구태원(Tae-Won Goo),윤은영(Eun-Young Yun) 한국식품영양과학회 2013 한국식품영양과학회지 Vol.42 No.2

국산 및 중국산 갈색거저리 유충의 일반성분의 함량 분석결과, 갈색거저리의 동결건조 분말은 고함량의 단백질(50.32∼52.99%)을 포함하고 있었고, 중국산 분말이 약 2.67% 정도 국산에 비해 높은 단백질을 함유하고 있었다. 갈색거저리의 필수 및 비필수아미노산을 포함하고 있었고, 분석된 아미노산의 함량은 국산과 중국산이 비슷한 수준을 나타내었으며, 비필수아미노산 중 glutamic acid의 함량이 다른 아미노산에 비해 가장 높게 존재하였다. 지방산의 분석 결과, 갈색거저리 분말은 총 지방산 중 76.80∼80.55%의 많은 불포화지방산을 함유하고 있었다. 불포화지방산 중 linoleic acid는 국산 20.80±1.1%, 중국산 34.69±1.9%로 13.89% 차이, oleic acid는 국산 51.40±0.47%, 중국산 40.20±1.5%로 11.2%의 차이, linolenic acid는 국산 0.5%, 중국산 1.4%로 명확한 함량 차이와 유의성(p<0.05)이 인정되었다. 유해물질인 식중독균(E. coli O157:H7, Salmonella spp.)은 국산과 중국산 분말 모두에서 검출되지 않았으며, 중금속 검사 결과에서 수은(Hg)이 국산과 중국산에서 각각 0.03 mg/kg, 0.08 mg/kg으로 검출되었으나 일반식품의 기준치 이하로 확인되었고 그 밖의 중금속은 존재하지 않았다. As part of a study on insects as food, the nutritional and harmful components in the mealworm (Tenebrio molitor) were analyzed. In addition, due to a recent introduction of live Chinese mealworms in the Korean market, components between Korean and Chinese mealworms were compared. Analysis of general composition (moisture, crude protein, crude fat, crude ash, crude fiber, and carbohydrates) showed that crude protein (50.32~52.79%) was abundant in both Korean and Chinese mealworm powders, with the protein content in the Chinese mealworm powder higher than that in the Korean mealworm powder by 2.67%. The amino acid compositions were similar, but the fatty acid compositions differed in the Korean and Chinese mealworm powders. The unsaturated fatty acid contents were 76.80~80.55% of the total fatty acid content in the mealworms. The linoleic acid contents in the Korean and Chinese mealworms were 20.8±1.1% and 34.69±1.9%; the linolenic acid contents were 0.47% and 1.31%; and the oleic acid contents were 51.40±0.9% and 40.20±1.5%, respectively. With respect to harmful components, such as heavy metals and bacteria that cause food poisoning, bacteria such as Escherichia coli O157:H7 and Salmonella spp. were not detected in both Korean and Chinese mealworms, and the mercury content was below the standard values for common foods (Korea, 0.03 mg/kg; China, 0.08 mg/kg).

조리방법별 갈색거저리 유충의 물리적 및 관능적 특성

백민희,윤영일,김미애,황재삼,구태원,윤은영,Baek, Minhee,Yoon, Young-Il,Kim, Mi Ae,Hwang, Jae-Sam,Goo, Tae-Won,Yun, Eun-Young 한국식품조리과학회 2015 한국식품조리과학회지 Vol.31 No.5

Recently, the Tenebrio molitor larva was recognized as a novel food ingredient by the Ministry of Food and Drug Safety in Korea. Accordingly, we investigated its physical and sensory characteristics to establish the cooking conditions that may increase the demand of T. molitor larvae as a food. In this study, T. molitor larvae were cooked by various methods such as hot air dry, oven-broil, roast, pan fry, deep fry, boil, steam, and by microwave. In the physical evaluation of texture, the hardness and fracturability values were highest when larvae were cooked in the microwave. The adhesiveness, springiness, and chewiness values were highest when larvae were boiled. Boiled and steamed larvae had the highest lightness (L value), while oven-broiled larvae had the highest redness (a value) and yellowness (b value) values. Sensory evaluations assessed the appearance, aroma, flavor, and texture of cooked T. molitor larvae. Steamed and boiled larvae sizes were significantly large and the form was well preserved similar to fresh larvae. The moisture heat cooked (steamed and boiled) T. molitor larvae had the aroma and flavor of steamed corn, canned pupa, and boiled mushroom. In case of oven-broiled T. molitor larvae, the aroma and flavor of mealworm oil, seafood, sweet and roasted sesame were higher than in those cooked by other methods. In texture among sensory evaluation, the hardness and crispiness were the highest in the hot air dried and oven-broiled larvae, whereas juiciness was significantly higher in the boiled and steamed. Accordingly, we suggest that oven-broiled T. molitor larva will be prefered by consumer, due to its the rich aroma and flavor.

쌍별귀뚜라미 식품 용도 개발에 따른 절식조건

안미영 ( Mi Young Ahn ),황재삼 ( Jae Sam Hwang ),윤형주 ( Hyung Joo Yoon ),박꽃하얀 ( Kyothayan Park ),김성현 ( Sunghyun Kim ),김은미 ( Eun Mi Kim ) 한국잠사학회 2015 한국잠사곤충학회지 Vol.53 No.2

본 연구는 사육용 쌍별귀뚜라미의 배변제거를 위한 절식시간을 알아보는 것을 실험 목적으로 하였다. 귀뚜라미는 현재 드디어 한시적 식품인정을 받았다. 따라서 식용 곤충으로서 국내시판을 위한 연구의 일환으로 귀뚜라미 생체 안의 배변을 제거하여 미생물의 번식을 예방하고 이취 제거와 맛을 향상시키기 위하여 본 쌍별귀뚜라미의 절식 실험을 실시한 결과 농가와 실험실에서 두 곳 모두 절식 2일째부터는 귀뚜라미의 습성에 의해 동족포식의 카니발리즘이 발동하여 귀뚜라미의 생체가 훼손되어 나감을 관찰 하였다. 쌍별 귀뚜라미 장내 분변을 현미경으로 살펴본 결과 1일째 이후 분변은 거의 없어지고 싸움으로 획득한 귀뚜라미의 팔다리 일부가 장내에 들어 있는 양상을 보여 1일이 귀뚜라미 장내 변이 거의 없어지는 시간임을 알 수 있어 생체 운송 배달시 1일이 걸리는 기간을 감안하면 절식시간은 1일이 적당하다고 사려 되어진다. This study aimed to determine a feed withdrawal duration time of Gryllus bimaculatus (Gb), for removal of intestinal feces. Finally, this species of cricket has been temporarily approved for use as a human food source in Korea. To commercialize Gb as an edible insect, we attempted to prevent the proliferation of micro-organisms and improve taste of Gb by removing intestinal feces of live crickets. Our study showed that weak crickets were damaged in 2 days after feed withdrawal in farm. The microscope findings also revealed that feces of strong crickets`` intestine almost disappeared at 1 day after feed withdrawal and instead, some of arms and legs of weak crickets were observed in the former``s intestine. The results suggest that 1 day is deemed to be an adequate time for feed withdrawal, when it usually takes 1 day to deliver live crickets from the farm to a destination.