보고서 정보
주관연구기관 |
국립농업과학원 National Institute of Agricultural Sciences |
보고서유형 | 최종보고서 |
발행국가 | 대한민국 |
언어 |
한국어
|
발행년월 | 2015-02 |
과제시작연도 |
2014 |
주관부처 |
농촌진흥청 Rural Development Administration(RDA) |
등록번호 |
TRKO201500010484 |
과제고유번호 |
1395035640 |
사업명 |
국책기술개발 |
DB 구축일자 |
2015-07-11
|
DOI |
https://doi.org/10.23000/TRKO201500010484 |
초록
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Ⅳ. 연구개발결과
○ 장수풍뎅이 유충 식용화를 위한 분말제조조건으로 절식, 세척, 멸균, 세척(멸균수), 동결 건조 설정하였고, 장수풍뎅이 유충 동결건조 분말은 전체 조단백 함량은 38.6%이며 아미노산(필수 33.6%, 비필수 57%) 중 Glutamic acid가 가장 함량이 높았고, 조지방 함량은 28.76%이며 이 중 올레산이 가장 높은 46.42% 차지하였음
○ 염증 유도한 대식세포에서 장수풍뎅이 처리시 3000 μg/ml에서 TNF-α는 약 14배, IL-6는 약 2배 억제, NO와 COX-2는 완벽히 억제하여
Ⅳ. 연구개발결과
○ 장수풍뎅이 유충 식용화를 위한 분말제조조건으로 절식, 세척, 멸균, 세척(멸균수), 동결 건조 설정하였고, 장수풍뎅이 유충 동결건조 분말은 전체 조단백 함량은 38.6%이며 아미노산(필수 33.6%, 비필수 57%) 중 Glutamic acid가 가장 함량이 높았고, 조지방 함량은 28.76%이며 이 중 올레산이 가장 높은 46.42% 차지하였음
○ 염증 유도한 대식세포에서 장수풍뎅이 처리시 3000 μg/ml에서 TNF-α는 약 14배, IL-6는 약 2배 억제, NO와 COX-2는 완벽히 억제하여 정상수준으로 회복함을 확인하였고, 염증 유도한 마우스의 혈청과 장기에서 장수풍뎅이 3000 mg/kg 경구투여 시 TNF-α는 약 2.7배, IL-6는 약 2.3, IL-1β는 약 2배, NO는 약 15배, COX-2는 약 2배 억제됨을 확인하였음
○ 장수풍뎅이의 식용화를 위해 장수풍뎅이 분말 100 g당 올레산(oleic acid)이 16.15 g으로 가장 높은 함량을 차지하므로 올레산을 대표물질로 선정함
○ 장수풍뎅이 분획물 및 대표지방산의 in vitro 항산화능, BACE1 저해활성 및 Aβ 유도 독성으로부터의 신경세포보호효과를 확인하였으며 순수물질인 올레산 및 감마 리놀렌산의 NF-κB 신경전달 체계 관련 항염증 반응을 통한 알츠하이머성 치매 예방 메커니즘을 규명함
○ DEN으로 유도된 간독성 동물모델에서 간세포의 죽음을 억제하여 지방간, 간경화를 완화시키는 간보호 활성을 규명하였고, 유충분획물들이 가진 암세포 특이적으로 작용하는 세포독성 활성을 규명함
○ 국제적 GLP 수준으로 유전독성 (복귀돌연변이, 체외염색체이상, 체내소핵시험) 및 일반 독성 (단회 및 4주, 13주 반복 경구투여 독성시험) 평가를 통해 DNA 또는 염색체 손상여부 및 경구투여에 따른 독성발현 여부를 확인하여 인체에 무해함을 체계적이고 과학적으로 증명함
○ 장수풍뎅이의 산업화를 위한 멸균공정, 동결건조 공정에 대한 제조공정 표준 확립 및 시제품 제조(제형, 포장용기, 재질, 포장방법, 표기사항)
○ 장수풍뎅이 시제품에 대한 가속실험을 실시하여 지표물질의 변화를 관찰하고 아레니우스 방정식에 의한 유통기한 설정
Abstract
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In this study, we established optimal powder manufacturing process for Allomyrina dichotoma larva for using as a novel food. For this purpose, it should be feed with the oak sawdust sterilized by water vapor for 3-5 days, starved without water or food for 3 days, and then the larvae were sterilized
In this study, we established optimal powder manufacturing process for Allomyrina dichotoma larva for using as a novel food. For this purpose, it should be feed with the oak sawdust sterilized by water vapor for 3-5 days, starved without water or food for 3 days, and then the larvae were sterilized before freeze-drying. The sterilized A. dichotoam larva were lyophilzed and grinded by a blender. A safety of the powder as a food was validated by evaluation of Raw 264.7 macrophage cytotoxicity using MTS assay. As above results, we propose that optimal powder manufacturing process established in this study can be used in industrial production of A. dichotoma as a novel food.
To evaluate A. dichotoma as food materials, we investigated their composition of nutritional and harmful components. Analysis of nutritional composition (moisture, crude protein, crude fat, crude ash, crude fiber and carbohydrates) showed that the content of crude protein, fat, carbohydrate were by 38.36%, 28.76%, 26.23% in A. dichotoma powder. In A. dichotoma, all bacteria were not detected, but small amount of Hg (0.06 mg/kg) was detected. And, we also investigated whether their extract have anti-inflammatory, anti-obesity, or anti-diabetic effect on various cell lines and mice.
Alzheimer’s disease (AD) is an progressive neurodegenerative disorder characterized by neuronal loss and extracellular senile plaques containing β-amyloid peptide (Aβ). Aβ is proteolytically produced by β- and γ-secretase from a single transmembrane amyloid precursor protein (APP). The progressive accumulation of insoluble Aβ peptide is considered to initiate a pathogenic cascade of AD. Considerable evidence has been suggesting that the reactive oxygen species (ROS) are involved in the apoptotic mechanism and inflammatory response of Aβ-mediated neurotoxicity. The present study was designed to demonstrate the anti-AD effect of A. dichotoma larvae extracts and their major compounds through the regulation of in vitro BACE1, Aβ-induced neurotoxicity and Aβ-induced inflammatory response. The isolated major compounds showed novel neuroprotective properties not only via the selective and specific inhibition of BACE1 activity but also through the alleviation of Aβ25-35-induced toxicity. Moreover, the results implied that extracts and their isolated compounds attenuated neuroinflammatory responses through the inactivation of NF-κB by NF-κB dependent inflammatory pathways and phosphorylation of MAPK in Aβ25-35-induced PC12 cells.
Beetle larvae have been used as a traditional medicine to treat various human liver diseases. To prove liver protective function of A. dichotoma larvae (ADL), we induced liver damages by intraperitoneal injection of a hepatotoxic reagent, diethylnitrosamine (DEN), to C3H/HeN male mice, and orally administered freeze dried ADL powder. ADL powder lessened DEN-induced hepatotoxicity considering reduced signs of acute and chronic hepatotoxicities such as ALP level in blood serum, TUNEL positive hepatocytes, ductural reaction, steatotic hepatocytes, and collagen deposition of the Masson’s trichrome staining. In addition to the hepatoprotection, anticancer activity of ADL has been examined. The ADL powder was extracted with ethanol and then fractionated with hexane, ethyl acetate, and water by solvent partition technique. The ethyl acetate fraction showed cytotoxicity to various cancer cells through induction of apoptosis and necrosis, and perturbed metabolism of the cancer cell to trigger autophagy. Collectively, ADL contains bioactive substances that can protect hepatocyte from toxic chemicals and trigger cell death in cancer cells. Thus, further purification and analyses of ADL fractions could lead to identification of novel bioactive compounds.
To Evaluate toxicological information of A. dichotoma for development as a food ingredient, we conducted the toxicity tests of the edible insects under the GLP condition. The safety and toxicity assessment for edible insects will contribute to secure the food source in the future. This research is significant to secure the initiating technology in the relevant filed. We carried out the establishment of validation method, the general toxicity studies, and the genotoxicity studies for freeze-dried powder from A. dichotoma. As a result, the validation method for the homogeneity, contents and stability analysis of freeze-dried powder from A. dichotoma was established. In the results of acute, 4 weeks and 13 weeks toxicity studies of freeze-dried powder from A. dichotoma, there were no toxicity findings in any studies. In the results of Ames, chromosomal aberration, micronucleus and comet assay, there were negative findings or no toxicity finding in any studies. The safety of A. dichotoma was established and these insects will be available for the food ingredient.
Freeze-dry condition (time, temperature, vacuum level) and sterilization condition (time, temperature) setting in insect for standardization of production process. Also, setting self standard test (appearance, moisture, foreign substance, crude protein, crude fat content etc.) with standardization of production process based on the Korea Food Standards Codex.
Check self standard test and material change on the acceleration test with regard to insect resource. And expiration date set at by the Arrhenius equation. Also, Finished producing a prototype of powder form with package design considering product characteristic and Check self standard test and material change on the acceleration test with regard to prototype. Based on these results a plan to apply for temporary registration with the Food and Drug processed foods
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