초록 ▼

① 강력한 항균활성 GRAS 미생물 개발 : 기존의 GRAS 등급 미생물 유래 항균제중 가장 강력한 항균력 및 항균 spectrum 지님
→ GRAS 등급 미생물 유래 항세균/항진균활성 물질(국내․외 최초)

② 천연항균물질 개발 : 분리·정제 과정 없이 유산균 생산 후 배양상징액 만으로도 식중독/부패균 제어 가능
→ 생산단가 낮음(process의 단축)

③ 식품원료규격의 천연항균물질 개발 : GRAS 등급 미생물(유산균: 식품원료 규격), 균주생산배지(식용배지: 식품 원료 규격) 모두 식품첨가물

① 강력한 항균활성 GRAS 미생물 개발 : 기존의 GRAS 등급 미생물 유래 항균제중 가장 강력한 항균력 및 항균 spectrum 지님
→ GRAS 등급 미생물 유래 항세균/항진균활성 물질(국내․외 최초)

② 천연항균물질 개발 : 분리·정제 과정 없이 유산균 생산 후 배양상징액 만으로도 식중독/부패균 제어 가능
→ 생산단가 낮음(process의 단축)

③ 식품원료규격의 천연항균물질 개발 : GRAS 등급 미생물(유산균: 식품원료 규격), 균주생산배지(식용배지: 식품 원료 규격) 모두 식품첨가물 규격이 아닌 식품원료 규격에 부합하도록 하여 국내․외 시장 진입 바로 가능
→ 실용화 가능

④ 균주 생산배지를 배추농가에서 과잉 생산된 배추, 김치제조사에서 김치생산 과정 중 발생하는 폐절임 배추 활용 방안 도출
: 미이용자원 및 폐자원의 자원화 및 부가가치 제고
→ 농가소득 증대 및 김치제조사 현안 해결
※ 본 연구성과물인 미생물유래 천연항균제는 기술이전 및 사업화

(출처 : 요약서 3p)

Abstract ▼

Purpose&Contents
1) Unit 1. Development of valuable microorganisms harboring antimicrobial activity against food-borne pathogens and application technique of new biopreservative into non-thermal processed foods

(1) Development of valuable microorganisms harboring antimicrobial activity aga

Purpose&Contents
1) Unit 1. Development of valuable microorganisms harboring antimicrobial activity against food-borne pathogens and application technique of new biopreservative into non-thermal processed foods

(1) Development of valuable microorganisms harboring antimicrobial activity against food-borne pathogens
- Isolation & identification of GRAS m/o harboring antimicrobial activity against food-borne pathogens
- Characterization of the developed natural antimicrobial agent
- Antimicrobial activity and spectrum against food-borne pathogens
- Growth inhibition mechanism of food-borne pathogens
- Purification and structural identification of the antimicrobial agents

(2) Application technique of new antimicrobial agents(biopreservative) into non-thermal processed foods.
- Technique for commercial production: Development of edible media for natural antimicrobial agent production.
- Production of natural antimicrobial agent in edible media
- Validation of the developed antimicrobial agent in non-thermal processed foods compared to commercially used natural preservative
- Application of the developed antimicrobial agent into food system(Lab scale: non-thermal processed food/fresh food/minimal processed food)

2) Unit 2. Production and commercialization of natural antimicrobial agent
- Set-up of process standardization of each non-thermal process food for commercialization
- Mass production of natural antimicrobial agent and development of purification tehchnique
- Development of application technique of the natural antimicrobial agent into non-thermal processed food system
- Development of profit creation model using the natural antimicrobial agents

3) Unit 3. Toxicity evaluation of antimicrobial agent derived from microorganism for control of food-borne pathogens in non-thermal processed food(Good laboratory practice)
- Evaluation target : Developed antimicrobial agents in this study
(antimicrobial agents derived from lactic acid bacteria
: 2 ea / derived from Bacillus subtilis : 1 ea)

Results
1) Unit 1. Development of valuable microorganisms harboring antimicrobial activity against food-borne pathogens and application technique of new biopreservative into non-thermal processed foods

① Development of GRAS m/o harboring antimicrobial activity against food-borne pathogens
: LAB 3 sp., Bacillus subtilis 1 sp.
→ The developed 3 LAB species showed strong antibacterial activities as well as antifungal activities.

② Purification and characterization of natural antimicrobial agents
ⅰ) Antimicrobial agent from LAB: 3-hydroxy-5-dodecenoic acid
→ The identified antifungal compounds world-widely: only 25 compounds Our report is only one report in Korea.

ⅱ) Antimicrobial agent from B. subtilis: a novel bacteriocin
→ Its a.a. sequence was registrated in NCBI; Acession NO. PRJNA309888; The novel bacteriocin was designated as Mejucin.

③ Development of edible media for LAB & Bacillus sp.
: Four kinds of edible media was developed using waste Korean cabbage.

④ Verification of comparative advantage compared to commercially used natural preservative
: The developed antimicrobial agent culture supernatant of GRAS m/o in this study showed much stronger activities as well as broader spectrum than the commercially used proservatives(4 kinds) even at their maximum recommended concentration.

⑤ Application of the developed antimicrobial agents into foods(non-thermal processed food/fresh food)
: 100% control of Food-borne pathogens/putrefactive microorganism However, the agents were not effective into useful fermentation m/o as well as olfactory characteristics of the foods.

2) Unit 2. Production and commercialization of natural antimicrobial agent
① Setup of standards processing for non-heated products for industrialization
: Standards processing and specifications for kimchi and pickles

② Mass production and purification technology of natural antibacterial materials
: Establishment of optimal culture conditions and mass production through concentration
→ Establishment of mass production process over 3 tons

③ Technology development of natural antibacterial material on non-heated food (industrial scale)
: Construction of non-heated food application 4 cases (kimchi, geotjeori, white radish cabbage, korean lettuce kimchi)

④ Development of creating profit models using new natural antibacterial materials.
: Check of replaceability through comparative antibacterial activity experiment of four kinds of existing antibacterial materials
: Establishment of case through application experiment of instant food
: Establishment of case for sensory improvement through deregulation of heating process
: Possession of data for extension of shelf life

3) Unit 3. Toxicity evaluation of antimicrobial agent derived from microorganism for control of food-borne pathogens in non-thermal processed food(Good laboratory practice)

① The toxicity study of crude antifungal compounds produced by Lactobacillus plantarum AF1+HD1
◦ Single dose oral toxicity study of in rat
The crude antifungal compounds produced by Lb. plantarum AF1+HD1 did not observed any toxic effects in single dose oral treated animals at 2000 mg/kg body weight dose level. Approximate lethal dose(ALD) of the material was considered to be higher than 2000 mg/kg body weight in rats.

◦ Repeated dose 4 weeks oral dose range finding study in SD rats(Non-GLP)
Adverse effect related with the dose of The crude antifungal compounds produced by Lb. plantarum AF1+HD1 was not observed up to 2000 mg/kg. Therefore, the NOAEL (No Observed Adverse Effect Level) of The crude antifungal compounds produced by Lb. plantarum AF1+HD1 was considered to be 2000 mg/kg/day (the highest dose tested) in both sexes. So dose range of 13 weeks repeated oral dose toxicity test was considered to be 0, 500, 1000, and 2000 mg/kg body weight.

◦ Bacterial reverse mutation study
The crude antifungal compounds produced by Lb. plantarum AF1+HD1 was not mutagenic (negative) under the conditions employed in the present study.

◦ In vitro mammalian chromosomal aberration test
The crude antifungal compounds produced by Lb. plantarum AF1+HD1 was not considered to have the ability to induce the chromosomal aberrations in CHL/IU cells under the present experimental conditions.

◦ Mammalian erythrocyte micronucleus test
The crude antifungal compounds produced by Lb. plantarum AF1+HD1 was determined not to induce an increased frequency of micronulei in the bone marrow cells of male ICR mice under the present experimental condition

② The toxicity study of crude antifungal compounds produced by Bacillus subtilis SN7
◦ Single dose oral toxicity study of in rat
The crude antifungal compounds produced by B. subtilis SN7 did not observed any toxic effects in single dose oral treated animals at 2000 mg/kg body weight dose level. Approximate lethal dose(ALD) of the material was considered to be higher than 2000 mg/kg body weight in rats.

◦ Repeated dose 4 weeks oral dose range finding study in SD rats(Non-GLP)
Adverse effect related with the dose of The crude antifungal compounds produced by B. subtilis SN7 was not observed up to 2000 mg/kg. Therefore, the NOAEL (No Observed Adverse Effect Level) of The crude antifungal compounds produced by B. subtilis SN7 was considered to be 2000 mg/kg/day (the highest dose tested) in both sexes. So dose range of 13 weeks repeated oral dose toxicity test was considered to be 0, 500, 1000, and 2000 mg/kg body weight.

◦ Bacterial reverse mutation study
The crude antifungal compounds produced by B. subtilis SN7 was not mutagenic (negative) under the conditions employed in the present study.

◦ In vitro mammalian chromosomal aberration test
The crude antifungal compounds produced by B. subtilis SN7 was not considered to have the ability to induce the chromosomal aberrations in CHL/IU cells under the present experimental conditions.

◦ Mammalian erythrocyte micronucleus test
The crude antifungal compounds produced by B. subtilis SN7 was determined not to induce an increased frequency of micronulei in the bone marrow cells of male ICR mice under the present experimental condition.

③ The toxicity study of crude antifungal compounds produced by Lactobacillus plantarum EM
◦ Single dose oral toxicity study of in rat
The crude antifungal compounds produced by Lb. plantarum EM did not observed any toxic effects in single dose oral treated animals at 2000 mg/kg body weight dose level. Approximate lethal dose(ALD) of the material was considered to be higher than 2000 mg/kg body weight in rats.

◦ Repeated dose 4 weeks oral dose range finding study in SD rats (Non-GLP)
Adverse effect related with the dose of The crude antifungal compounds produced by Lb. plantarum EM was not observed up to 2000 mg/kg. Therefore, the NOAEL (No Observed Adverse Effect Level) of The crude antifungal compounds produced by Lb. plantarum EM was considered to be 2000 mg/kg/day (the highest dose tested) in both sexes. So dose range of 13 weeks repeated oral dose toxicity test was considered to be 0, 500, 1000, and 2000 mg/kg body weight.

◦ Bacterial reverse mutation study
The crude antifungal compounds produced by Lb. plantarum EM was not mutagenic (negative) under the conditions employed in the present study.

◦ In vitro mammalian chromosomal aberration test
The crude antifungal compounds produced by Lb. plantarum EM was not considered to have the ability to induce the chromosomal aberrations in CHL/IU cells under the present experimental conditions.

◦ Mammalian erythrocyte micronucleus test
The crude antifungal compounds produced by Lb. plantarum EM was determined not to induce an increased frequency of micronulei in the bone marrow cells of male ICR mice under the present experimental condition.

Expected Contribution

(출처 : SUMMARY 8p)

목차 Contents

• 표지 ... 1
• 제 출 문 ... 2
• 보고서 요약서 ... 3
• 요 약 문 ... 5
• SUMMARY ... 8
• CONTENT ... 13
• 목차 ... 15
• 1. 연구개발과제의 개요 ... 17
• 제 1 절 연구개발의 목적 ... 17
• 제 2 절 연구개발의 필요성 ... 17
• 제 3 절 연구개발의 범위 ... 20
• 2. 국내외 기술개발 현황 ... 26
• 제 1 절 국내외 기술개발 및 시장(제품) 현황 ... 26
• 제 2 절 본 연구결과가 국내외 기술개발 현황에서 차지하는 위치 ... 32
• 3. 연구수행 내용 및 결과 ... 34
• 제 1 장 제1세부 항식중독 미생물 활성을 지닌 유용 미생물의 개발 및 비가열 식품에 신규 천연항균소재 적용 기술 개발 ... 34
• 제 1 절 연구수행 방법 ... 34
• 제 2 절 연구내용 및 결과 ... 54
• 제 2 장 제1협동 천연항균소재의 산업화 및 상품화 ... 214
• 제 1 절 연구수행 방법 ... 214
• 제 2 절 연구내용 및 결과 ... 217
• 제 3 장 제2협동 비가열 식품의 식중독 제어를 위한 미생물유래 천연항균제의 독성평가 ... 251
• 제 1 절 연구수행 방법 ... 251
• 제 2 절 연구내용 및 결과 ... 254
• 제 4 장 종합분석 및 고찰 ... 307
• 제 1 절 연구결과 종합 요약 ... 307
• 제 2 절 기술적 성과 ... 327
• 제 3 절 경제적 성과 ... 329
• 제 4 절 교육지도 ･ 홍보 성과 ... 331
• 제 5 절 학술발표 ･ 인력양성 성과 ... 333
• 4. 목표달성도 및 관련분야 기여도 ... 335
• 제 1 절 목표달성도 ... 335
• 제 2 절 관련분야 기여도 ... 337
• 5. 연구결과의 활용계획 ... 345
• 제 1 절 실용화 ･ 산업화 계획 ... 345
• 6. 연구과정에서 수집한 해외과학기술정보 ... 347
• 7. 연구개발결과의 보안등급 ... 352
• 8. 국가과학기술종합정보시스템에 등록한 연구시설･장비 현황 ... 353
• 9. 연구개발과제 수행에 따른 연구실 등의 안전조치 이행실적 ... 354
• 10. 연구개발과제의 대표적 연구실적 ... 358
• 11. 기타사항 ... 360
• 12. 참고문헌 ... 361
• 끝페이지 ... 366