초록 ▼

○ 연구결과
1.[제1세부]김치발효 미생물 천이조절 우량 종균의 개발과 미생물 천이조절 김치의 개발
가. 실용화 천이조절 종균 2종이상 개발
- 본 과제에서는 위의 결과를 토대로 김치종균의 요건,즉 ①천이조절기능 + ②발효김치맛의 우수능 +③기능성 +④산업적 내구성을 지닌 종균을 최종 4종 개발함.
나. 종균 우점율 70%이상,산도 1%이하가 4개월 유지
- 본 연구 결과로부터 종균 우점 70%,산도 1%이하,4개월 유지가 성공적으로 이루어짐. 현행 김치저장기간(냉장에서)28일 → 김치저장기간 4개월로 연

○ 연구결과
1.[제1세부]김치발효 미생물 천이조절 우량 종균의 개발과 미생물 천이조절 김치의 개발
가. 실용화 천이조절 종균 2종이상 개발
- 본 과제에서는 위의 결과를 토대로 김치종균의 요건,즉 ①천이조절기능 + ②발효김치맛의 우수능 +③기능성 +④산업적 내구성을 지닌 종균을 최종 4종 개발함.
나. 종균 우점율 70%이상,산도 1%이하가 4개월 유지
- 본 연구 결과로부터 종균 우점 70%,산도 1%이하,4개월 유지가 성공적으로 이루어짐. 현행 김치저장기간(냉장에서)28일 → 김치저장기간 4개월로 연장 성과
다. 천이조절 김치 개발이 실험실 recipe김치뿐만 아니라 참여업체 김치에서도 성공적으로 이루어짐.
- 본 연구개발에서는 본 실험실에서 사용한 김치원+부재료하에서의 김치에서 뿐만 아니라,참여기업 3사의 김치환경하에 개발종균을 적용하여 실용화 가능성을 높임.위 표에서 보여지는 바와 같이 참여기업 3사중 H사 GR1종균김치에서는 4개월 김치저장에도 산도 1%이하 종균우점 72%이상 유지에 성공함.
이에 반해 종균을 접종하지 않은 H사김치는 산도 1.13%이고,이때 우점 유산균은 99.2%가 homoferment ative type인 Lactobacillussp.로 밝혀짐.
H사 GR1종균김치는 200인 이상 일반 소비자 관능평가에서 전체적 기호도 항목에서 4.07점(5점만점)으로 ‘우수’함으로 평가됨.
2. [제2세부]수출김치의 산막효모 제어 기술의 개발
가. 산막효모에 대해 강력한 항균활성을 지니는 김치유산균,고초균이 확보
- 김치유산균 3종 :Lb.plantium AF1,NO1,HD1 고초균 2종 :B.velezensisS100-2,S100-100
나. GRAS미생물로부터 실용화 가능한 산막효모제어용 항진물질 확보
- 항산막활성 원인 물질 규명 :
① 5-oxododecanoic acid, 3-hydroxydecanoic acid, 3-hydroxy-5-dodecanoic acid from Lb. platarum HD1
② δ-Dodecalactone, cyclo-Leu-Leufrom Lb. platarum AF1
→ 세계 12번째 유산균 유래 항진물질 규명,국내최초
다. 개발식용배지에서 생산된 항진균물질 처리에 의하여 실제 상품김치에서 산막효모가 2개월이상 불검출
- 시판김치에서 사용하고 있는 가스흡입제가 부착된 retortpouch1kg포장재에 김치를 담고,개발 유산균 항진물질 처리구와 대조구①로 항진물질 무처리구,대조구②로 김치산업체에서 현재 가장 널리 사용하고 있는 유까추출물과 자몽추출물을 처리함.
- -1℃에서 3개월 저장시 대조구①인 무처리구에서는 62일 만에 산막효모가 검출되기 시작하였고, 대조구②에서는 74~91일 만에 산막효모 검출됨.이에 반해 개발유산균의 처리구는 3개월 이상 저장에도 산막효모가 불검출 되어 우수한 활성 나타냄.
※ 산막효모의 김치에서 영향 :군덕내(이취),연부
☞ -1℃에서 3개월이상 저장김치에서 산막효모 불검출,기존 시판 산막효모제보다 강력한 활성 보임.
3.[제3세부]동물실험을 통한 개발제품의 독성 및 기능성 평가
가. 산막효모제어균인 Lactobacillus plantarum AF1과 그 배양액의 독성평가
- 시료 2종 :Lb. plantarum AF1과 그 배양액
- 산막효모제어균인 Lb. plantarum AF1과 그 배양액 모두 단회 경구 투여 후 14일 간 관찰한 결과 저독성의 안전한 물질로 확인되었음.
- 산막효모제어균인 Lb. plantarum AF1과 그 배양액 모두 4주간 반복 경구 투여 후 시험물질에 기인하는 이상 소견을 발견할 수 없는 것으로 보아 저독성의 안전한 물질로 확인되었음.
- 산막효모제어균인 Lb.plantarum AF1과 그 배양액 모두 E.ColiPQ37에 대한 돌연변이 유발능(SOSchromotest)을 조사한 결과와 Chinesehamster유래 폐섬유아세포(CHL)를 이용하여 염색체 이상시험을 수행한 결과 유전 독성학적으로도 안전한 물질임.
나.개발김치 종균인 Leuconostoccitreum GR1과 그 배양액의 독성평가
- 시료 2종 :Leuc.citreum GR1와 그 배양액
- 개발김치 종균인 Leuc.citreum GR1과 그 배양액의 단회 경구 투여 후 관찰한 결과 저독성의 안전한 물질로 확인되었음.
- 개발김치 종균인 Leuc.citreum GR1과 그 배양액의 4주 반복 경구 투여 후 저독성의 안전한 물질로 확인되었음.
- 개발김치 종균인 Leuc.citreum GR1과 그 배양액 모두 유전 독성학적으로도 안전한 물질임.
다.김치유산균이 생성한 SPE정제 조항진균 물질의 안정성 농도 설정 및 독성평가
- 시료 2종 :Lb.plantarum AF1과 Lb.plantarum HD1SPE 정제 조항진균 물질
- 항진균 물질의 독성평가 농도 설정 :1차년도 과제에서 사용한 농도의 역가와 실제 개발김치 내농도를 고려하여 설정(김치 하루 섭취권장량 보다 100배 이상 사용)
- Lb. plantarum AF1과 Lb. plantarum HD1 SPE정제 조항진균 물질들을 단회 경구 투여 후 관찰한 결과 저독성의 안전한 물질로 확인되었음.
- Lb. plantarum AF1과 Lb. plantarum HD1 SPE 정제 조항진균 물질들을 4주 반복 경구 투여 후 저독성의 안전한 물질로 확인되었음.
라. 김치분말의 기능성평가
- in vitro항비만 효능 검증 : 3T3-L1지방전구세포와 돼지 지방전구세포의 증식과 분화를 농도의존적으로 억제하였으며,LPL활성 억제 및 중성지방 함량과 지방세포 내 중성지방의 축적을 억제하여 invitro항비만 효능을 검증하였음.
- in vivo항비만 효능 검증 : 고지방-고콜레스테롤식이 급여로 인하여 증가되어진 체중,부고환 지방조직,장간막지방조직 및 등지방조지의 무게가 특히 김치분말 10%첨가로 유의하게 저하되어 비만 억제효과가 있는 것으로 나타났으며,지방축적인자로 알려진 LPL 활성도 저하시켜 in vivo항비만 효능을 검증하였음.
4. [제1협동]DGGE기법을 통한 김치 및 원부재료의 미생물 천이 규명
가. 계절별 배추와 김치 부재료의 미생물상 분석
- 절임배추 겉잎과 속잎의 미생물상은 일치하였고 3개 회사 시료는 차이 보였으나 주요 미생물은 Leuconostoc, Lactobacillus, Weissella속이었음 .
- 세 회사 김치에 모두 존재한 미생물은 Leu.mesenteroides,회사별로 미생물상은 차이보였음.
- 파의 주요 미생물은 Weissella, 마늘의 주요 미생물은 Leuconostoc
나. 참여기업에서 수집한 김치의 계절별,제조사별,발효온도별 미생물상 분석
- 여름 제조 김치의 미생물상이 가장 다양하였고 (총 14종)모든 게절에 공통으로 존재한 미생물은 Leuconostoc, Lactobacillus, Weissella
- 제조사별 김치 미생물상은 서로 동일하였으나 분포는 달랐음.
- 발효 온도별 미생물상의 차이는 크지 않았고 미생물상의 변화속도만 달랐음.
다. 미생물 천이조절 김치와 일반김치 및 분말김치의 미생물상 분석
- 사용한 종균이 김치 저장 16주째까지 우점으로 존재하여 종균의 유용성이 확인 되었음.
- 일반김치에서는 Leuconostoccitreum은 존재하지 않았으며 천이조절 김치와 일반김치 모두 Lb. sakei가 종균을 제외한 젖산균들 중 3-4개월 숙성한 김치의 특징적인 균으로 생각됨.
- 분말김치의 분쇄조건별,저장기간 및 온도별 미생물상은 차이가 없었음.
5. [제2협동]종균 미생물의 대량 배양 및 안정화 기술 개발
가. 기존 MRS 대비 1/15이하 단가의 신규배지 개발
- 일반적으로 이용되고 있는 유산균 배지인 MRS 배지는 가격이 높아서 종균의 대량화를 위한 대체 배지 개발
- 폐배추 추출물은 잎 추출물 30%,당은 maltose 2%, 질소원은 yeastex tract 0.25%, 무기질은 2X salt stock(2% sodium acetate trihydrate,0.8% disodium hydrogen phosphate,0.8% sodium cirate,0.8% ammonium sulfate,0.04% magnesium sulfate,0.02% manganese sulfate)로 결정하였고,이 배지를 MFL배지로 명명함.
- 본 연구에서 신규배지를 300mL까지 배양시켰음.종균보급을 위한 대량배양 가능
- MRS 대비 2.2배 생균수 증가, MRS 대비 배지 가격 10.8배 감소,균수를 반영한 배지 가격 절감효과는 약 23배로 목표치 15배보다 더 큰 단가 절감 효과를 볼 수 있음.
나. 300L의 대량 배양에서 10⁸ CFU/mL이상의 생육에 도달
- Agitation speed는 50 RPM, pH control은 6.8,pH 조절제는 NaOH 용액을 이용하였고, inducer는 전체 발효액부피의 0.1%인 200ml을 첨가하여 30℃에서 발효를 실시함.
- 생균수 1.0×10¹⁰ CFU/ml에 도달하여 500L 발효조에 새로 개발된 배지를 300L 발효함으로써 최적화 성공
다.종균저장 시 생존율 80%이상, 3개월 이상 유지
- 종균의 안정화는 여러 가지 종류의 안정제를 첨가하거나 저장 시 저장온도를 결정하여 생존율을 높게 유지할 수 있는 시스템 개발
- 기존의 안정제에 관한 연구에서는 완충용액,소당류,trehalose,산화방지제 등을 이용하여 균의 저장성을 연장하였으나,본 연구 결과 0℃에서 기존의 안정제 보다 새로 개발한 배지인 MFL를 안정제로 첨가하였을 때 생존율이 더 높게 나타났으며 4주 동안 기존의 안정제는 30%의 생존율도 미치지 못하는데 비해 MFL은 4주까지 생존율 87.9%를 유지함.그러나 3개월간 80%에는 미치지 못함.
- 안정제로 MFL를 이용하고 glycerol5% 또는 10%를 첨가하여 균주를 -20℃에서 저장했을 때 3개월 동안 80% 이상의 생존율 유지
- 균주보관에 냉동건조 시키는 방법이 많이 이용되고 있지만,본 연구 결과 농축종균의 저온 저장이 보다 효과적이었음.
6. [제3협동]분말김치의 개발과 상품화
가. 분말김치의 개발
- 숙성김치(pH 4.0-4.3)에 동결건조보호제로 maltose 또는 lactose를 1.5% 첨가하여 동결건조한 후 powdermixer를 사용하여 14,000rpm에서 2분간 분쇄하는 조건으로 분말김치 개발
☞ 냉동(-20℃)에서 4개월 저장 시 생균수 109 CFU/g유지하여 6개월 이상 저장 가능
나. 분말김치의 제형화/제제화
- 분말김치의 제형화 및 제제화 조건을 수립하여 캡슐형,과립형,정제형 분말김치를 개발함

Abstract ▼

Unit 1. Development of a kimchi starter for control of microbial succession in kimchi and a microbial succession controling kimchi
A. Development of a kimchi starter for control of microbial succession in kimchi
a. Requisite for starter
① Bacteriocin production ability
② Bacteriocin prod

Unit 1. Development of a kimchi starter for control of microbial succession in kimchi and a microbial succession controling kimchi
A. Development of a kimchi starter for control of microbial succession in kimchi
a. Requisite for starter
① Bacteriocin production ability
② Bacteriocin production should be induced by presence of sencetive cells
③ Fermentation characteristics : good flavor and taste(mannitol production ability)
④ Functionality : safety, survive in a intestinal environment, adherence, anti-cancer effect ect.
⑤ Resistance to industrial environment : resistance to salt, acid, heat treatment and stability during microbial generation
☞ We develop four starters to satisty the above five requirements.
(Leu. citruem GR1, Leu. kimchii GJ22, Lb. sakei YY1, SC1)
b. Control of kimchi fermentation by using the starter(starter dominance rate 70↑, activity 1%↓, maintenance period 4 months)
◦ The results aresummerized as follows
① single starter
② mixed-starter
③ Sensory characteristics of the developed starter kimchi
☞ The results suggested that starter dominance rate 70%↑ and activity 1%↓ for four months were successfully achieved.
Present technical status 28 day atrefregerate temp
→ Kimchi storage period was extended to 4 months
Unit 2. Development of film-forming yeast control technique for export-kimchi
A. Development of GRAS microorganisms harboring anti-film-forming yeast activity
a. Identification of film-forming yeast(Gomori) in kimchi
☞ The yeasts associated with film-forming, off-odor and softening-texture were isolated and identified
- Pichia kudriavzevii 3 sp.
- Kazachstania servazzii 2 sp.
- Kazachstania bulderi 1 sp.
- Kazachstania exigua 1 sp.
b. From 108 species of kimchi lactic acid bacteria(LAB) and 100 species of Bacillus from fermented soybean products
◦ Development of strong anti-film-forming yeast activity kimchi LAB(3 species)
→ Lactobacillus plantarum AF1, Lactobacillus plantarum NO1, Lactobacillus plantarum HD1
◦ Selection of Bacillus sp. harboring the strongest activity against film-forming yeast
→ Bacillus velezensis SSH100-2, Bacillus velezensis SSH100-10
☞ Kimchi LAB 3 species : Lb. plantium AF1, NO, HD
Bacillus 3 species : B. velezensis SSH100-2, SSH100-10
c. Characterization of anti-fungal compounds(anti-film-forming yeast activity compounds)
◦ Identification of antifungal compounds from LAB
① 5-oxododecanoic acid, 3-hydroxydecanoic acid, 3-hydroxy-5-dodecanoic acid from Lb. platarum HD1
② δ-Dodecalactone, cyclo-Leu-Leu from Lb. platarum AF1
☞ ① & ② compounds were the 12th identification in the world from LAB. Those are the first identification in korea.
◦ Identification of antifungal compounds from Bacillus
① Iturin C14-C15
B. Development of technique for control of film-forming yeast on kimchi
a. Development of edible LAB medium
: LAB are cultivated in the edible medium and the culture extracts are used for anti-fungal compounds to prohibit the growth of film-forming yeast on kimchi
b. Treatments of the developed antifungal compounds from LAB
◦ Kimchi was packed in 1 kg retort pouch attached with agas-inhalant
◦ Treatment:
① antifungal compounds from Lb. plantarum AF1, Lb. plantarum HD1
② control ① : non-treatment
③ control ② : Yucca extract, Grapefruit extract
☞ The kimchitreated with AF1 and HD1 antifungal compounds did notshow any film-forming yeasts during 3 months at-1℃.
The developed antifungal compounds showed the strongest activity against film-forming yeasts.
Unit 3. Toxicity study and functional evaluation of the products developed through animal testing
The present study was carried out to investigate the in vivo single dose acute toxicity of Lactobacillus plantarum AF1(Lb. plantarum AF1)and its cell medium or Leuconostoc citreum GR1(Leuc. citreum GR1)and its cell medium, a lactic acid bacterium isolated from kimchi, in ICR male and female mice. The test articles were orally administered once to both sexes of mice. Mortalities, clinical findings, autopsy findings, and body weight changes were monitored daily for two weeks. Male and female mice were gavaged with Lb. plantarum AF1 and its cell medium or Leuc. citreum GR1 and its cell medium of four doses. No significant changes in general conditions, body weights, clinical signs and any gross lesions was observed in both sexes of mice administered orally with Lb. plantarum AF1 and its cell medium or Leuc. citreum GR1 and its cell medium. The results suggest that the no adverse effect was found in Lb. plantarum AF1 and its cell medium or Leuc. citreum GR1 and its cell medium.
This study was also performed to investigate the 4-week repeated-dose toxicity of Lb. plantarum AF1 and its cell medium or Leuc. citreum GR1 and its cell medium, a lactic acid bacteriums isolated from kimchi, in male and female rats. Sprague-Dawley male and female rats were divided into four groups, with 10 animals in each group. There were no test article-related deaths or abnormal clinical signs in either gender of rat during the observation period. Furthermore, no differences were found between the control and treatment groups in terms of body weight changes, food intake, and water consumptions. Hematological parameters, serum biochemical analysis, and any other findings also showed no significant or dose-dependent alterations. There were no alterations in organ weights upon administration of Lb. plantarum AF1 and its cell medium or Leuc. citreum GR1 and its cell medium. These results suggest that there were no adverse effects of oral application of Lb. plantarum AF1 and its cell medium or Leuc. citreum GR1 and its cell medium in both male and female rats.
We investigated the acute toxicity from a single dose of the crude anti-fungal compounds produced by Lb. plantarum AF1 or Lb. plantarum HD1 in ICR male and female mice in vivo.The test articles were orally administered once to both sexes of mice. The mortality rate, clinical findings, autopsy findings, and body weight changes were monitored daily for 14 days. In the oral acute toxicity test, male and female mice were gavaged with four doses (5, 50, 300 or 2,000m g/kg)of the crude an-ti-fungal compounds. The oral LD50 of the crude anti-fungal compounds was higher than 2,000 mg/kg. No significant changes in general conditions, body weights, clinical signs, or appearance of gross lesions were observed. In conclusion, our results suggest a low toxicity and no-adverse-effects from crude anti-fungal compounds produced by Lb. plantarum AF1 or Lb.plantarum HD1 up to 2,000mg/kg via the oral route.
This study was also performed to investigate the four-week repeated-dose toxicity of the crude antifungal compounds produced by Lb. plantarum AF1 or Lb. plantarum HD1 in male and female rats. Sprague-Dawley male and female rats were divided into four groups, with 10 animals in each group. The test articles were administered once daily by gavage to rats at dosage levels of 0,500, 1,000, and 2,000 mg/kg/day for four weeks. There were no test-article-related deaths or abnormal clinical signs in both the male and female rats during the observation period. Furthermore, no differences in the body weight changes, food intake and water consumption levels of the control and treatment groups were found. The hematological parameters, serum biochemical analysis results, histopathological examination results and all other findings also showed no significant or dose-dependent changes. There were also no changes in the organ weights upon the administration of the crude antifungal compounds produced by Lb. plantarum AF1 or Lb. plantarum HD1. These results suggest that the oral administration of the crude antifungal compounds produced by Lb. plantarum AF1 or or Lb. plantarum HD1 had no adverse effects up to a dosage level of 2,000 mg/kgin both male and female rats.
The study was carried out to determine the effects of the kimchi powder on proliferation and differentiation of 3T3-L1 preadipocytes and pig preadipocytes. To measure the cell proliferation and differentiation, the cells were treated with 25,50 and 100 μg/mL ethanol extracts of the kimchi powder for two days, and dimethyl sulphoxide(DMSO) was used as the control group. The kimchi powder extract showed the inhibitory action on proliferation, differentiation, triglyceride accumulation and LPL activity of 3T3-L1 preadipocytes and pigpreadipocytes in a dose-dependent manner. These actions indicate that the ethanol extracts of kimchi powderm ay have potential to reduce the fat accumulation and obesity.
This study was carried out to investigate the effect of kimchi powder on anti-obesity effects in rats fed a high-fat/high-cholesterol diet for 4 weeks. Weight-matched male Sprague-Dawley rats were assigned to four groups according to dietary fat, cholesterol levels and kimchi powder levels. Experimental groups were normal diet group(N), high-fat/high-cholesterol diet group (C), high-fat/high-cholesterol diet with 5% kimchi powder group (CKL) and high-fat/high-cholesterol diet with 10% kimchi powder group (CKH). The body weight gain and FER were increased by a high-fat/high-cholesterol diet, but significantly decreased in the CKH group, compared with the C group. The food intake was not different among the groups. The adipose tissues weights of C group were heavier than that of N group, whereas those of groups administered kimchi powder were gradually decreased. The serum ALT, AST, ALP and LDH activities elevated by a high-fat/high-cholesterol diet were significantly decreased by kimchi powder administration. Levels of serum total cholesterol and LDL-cholesterol tended to bedecreased in the kimchi powder fed groups compared with the C group. The serum HDL-cholesterol level decreased in the C group and markedly increased in the kimchi powder fed groups. Levels of triglyceride and total cholesterol in liver were lower in kimchi powder administered groups than in C group. These results suggest that kimchi powder may improve lipid metabolism and prevent obese.
Unit 4. Investigation of microbial succession of kimchi and itsingredient susing DGGE technique
Bacterial distributions in kimchi and kimchi ingredients were examined through the culture-independent PCR-DGGE technique. Salted Chinese cabbage samples, ingredient mix, and kimchi samples were collected from three kimchi companies in July, October, and January and were analyzed. Inner and outer leaves of salted Chinese cabbage were compared. Distributions of lactic acid bacteria (LAB) in garlic and green onion samples collected from kimchi companies and localold marketsin Korea were also analyzed by comparing the SDS-PAGE whole cell protein patternsand 16S rRNA sequences. Bacteria profile of kimchi samples made with starter culture were analyzed by PCR-DGGE and were compared to that of commercial kimchi samples during the fermentation and aging periods.There was little significant seasonal difference in bacterial distribution in salted Chinese cabbage and kimchi samples but slightly more varied bacteria profile in samples obtained in summer. A total of 360 LAB were isolated from 10 garlic and 7 green onion samples and were differentiated into groups by comparing SDS-PAGE whole cell protein patterns. Major LAB in garlic were Leuconostoc (Lc.), Weissella (W.), and Lactobacillus (Lb.) and in green onion were Weissella, Leuconostoc, and Lactococcus through the 16S rRNA sequence analysis. Leuconostoc was the most dominated LAB in garlic and Weissella was the most dominated LAB in green onion. LAB in salted Chinese cabbage were analyzed by both culture-dependent SDS-PAGE and culture-independent PCR-DGGE techniques followed by sequencing of the 16S rRNA gene. The results were compared to those of LAB that had previously been found in kimchi. The two identification methods produced distinct overal lLAB profiles. The PCR-DGGE method detected a more diverse microflora, including non-LAB strains. The culture-dependent method uniquely detected Weissella sp. and was able to provide the quantitative distribution of LAB in samples. Lc. mesenteroides, Lb. curvatus, and Lc. carnosum, which had also been reported as the dominantLAB in kimchi in the previous studies, were identified by both methods. Changes in bacterial microflora of two commercial kimchi, salted cabbage, and ingredient mix samples during 30 days of fermentation at 4℃ and 10℃ were monitored by PCR-DGGE technique. Leuconostoc was the dominant LAB over Lactobacillus at 4℃ despite the more varied LAB profile at 10℃. Weissella confusa was detected in the ingredient mix and also in samples throughout fermentation in both samples at 4℃ and 10℃. Lc. gelidum was detected as the dominant LAB at 4℃ in both samples. The temperature affected the LAB profile of kimchi by varing the pH, which was primarily caused by the temperature-dependent competition among different LAB speciesin kimchi. At 4℃, the sample variationsin pH and titratable acidity were more conspicuous due to the delayed growth of LAB. The LAB identified in kimchi ingredient samples in this study were previously found as dominant microorganisms in kimchi. The initial microflora in kimchi sample is probably determined mainly by the micr of loraofingredient mix, not by that of the salted cabbage.
Unit 5. Development of fermentation conditions for industrial application and stabilization technology
In the kimchi manufacturing process, the starter is cultured on a large-scaleand needs to be supplied at a low price to kimchi factories. However, current high costs associated with the culture of lactic acid bacteria or the starter, haveled to rising kimchi prices. To solve this problem, the development of a new medium for culturing lactic acid bacteria was studied. The base materials of a this novel medium consisted of Chinese cabbage extract, a carbon source, a nitrogen source, and inorganic salts. The optimal composition of this medium was determined to be 30% Chinese cabbage extracts, 2% maltose, 0.25% yeast extract, and 2X saltstock (2% sodium acetate trihydrate, 0.8% disodium hydrogen phosphate, 0.8% sodium citrate, 0.8% ammonium sulfate). The newly developed medium was named MFL (Medium For Lactic acid bacteria). After culture for 24hr at 30℃, the CFU/mL of Leuconostoc citreum GR1 in MRS and MFL was 3.41×109 and 7.49×109, respectively. The number of cells in the MFL medium was 2.2 times higher than their number in the MRS media. In a scale-up process using this of optimized medium, the fermentation conditions for Leuc. citreum GR1 were tested in a 2 L working volume using a 5 L jar fermentor at 30℃. At an impeller speed of 50 rpm(without pH control), the viable cell count was 8.60 ×109 CFU/mL. From studies on pH-stat control fermentation, the optimal pH and regulating agent was determined to be 6.8 and NaOH, respectively. At an impeller speed of 50 rpm with pH control, the viable cell count was 1.14 ×1010 CFU/ml after cultivation for 20 hr. This value was 3.34 times higher than that obtained using the MRS media in biomassp roduction. This MFL media is expected to have economic advantages for the cultivation of Leuc. citreum GR1 as a starter for kimchi production.
For a stable supply of kimchi starter, Leuconostoc citreum GR1 to kimchi industry, stabilizer, storage conditions and thermal stability of Leuc. citreum GR1 were analyzed. Trehalose, skim milk, tween 80, IMO, vitamin C, PBS solution and MFL were used as stabilizer for concentrated kimchi starter storage at 0℃, 4℃, 15℃, and 25℃. According to the results of 4 weeks of storage, 0℃ and MFL was most excellent as storage temperature and stabilizer, respectively. The survival rates after 4 weeks storage were MFL (87.9%) > 1X PBS solution (24.1%)> 5% tween 80 (16.4%) > 5% skim milk (13.9%)> 5% trehalose (0%) > 5% IMO (0%), which showed a significant difference in survival rate. Using the MFL as a primary stabilizer, glycerol was added to a concentration of 0%, 1%, 5%, and 10% and stored at 4℃, 0℃, and -20℃ for 12 weeks. Storage at –20℃ was the most good, and at –20℃ storage, glycerol concentrations of 5% or morecould be maintained more than 80% survival rate of Leuc. citreum GR1. Those results indicated that the MFL as stabilizer, -20℃ as storage temperature, and 5% or more concentrations of glycerol was the most effective for storing concentrated kimchi starter. Heat resistance of Leuc. citreum GR1 was determined in vitro. D-values at 46℃, 50℃, 54℃, and 58℃ ware 3,333min, 400min, 19min, and 1min. z-value of Leuc. citreum GR1 inferred from the Arrhenius equation was 3.44℃ and the activation energy was 586.46 KJ/mol.
Unit 6. Development and Commercialization of Kimchi Powder
A. Objectives and Significance of Research
- This study was conducted to development of formulation technology and commercialization of Kimchi powder to increase the shelf-life of Kimchi, and find a way to intake Kimchi without reluctance to foreigners who are not familiar with Kimchi.
B. Scope and Contents of Research
a.Development of Kimchi Powder
- Optimization of freeze-drying condition
- Establishment of micro-particle condition of Kimchi powder
- Analysis of physicochemical properties of Kimchi powder
b. Formulation of Kimchi Powder
- Diversification of formulated-products
- Selection of bonding-agent set by formulation and establishment of adding condition
- Establishment of expiration date of Kimchi powder
C. Results of Research
a. Development of Kimchi Powder
- Kimchi was fermented on 10℃ for 2 weeks, showing pH 4.0-4.3, to keep the 10⁷-10⁸ CFU/g level of lactic acid bacteria and then usedas as a mple for freeze-frying. Crushed Kimchi was more suitable for Kimchi powder than cuted Kimchi due to shortening the drying time. Also, set temperature of freeze-dryer shelf must be under 30℃ to keep the lactic acid bacteria. As a result of adding freeze-drying protect agent for improving survival ratio of lactic acid bacteria, lactose 1.5% showed higher survival rate 1 log CFU. Kimchi powder was micro-particulated using powder mixer, and then micro-particle condition was set 14,000 rpm for 2 min based on color, ASTA value, and sensory evaluation. Moisture content of Kimchi powder was 15.8%, and the number of lactic acid bactria showed 1.53 ×10⁷ CFU/g.
b. Formulation of Kimchi Powder
- Various of Kimchi powder was attempted such as capsule granula, and tablet for product diversification. Capsule-type product was manufactured using gelatin capsule filling 350 mg Kimchi powder. Granula-type product was carried out by mixing with ethanol, and then 4g of KImchi powder was granulated. Tablet-tyleproduct was produced with granulated Kimchi powder. Round and rectangle shape of tablets were manufactured to improve the amount of Kimchi powder per 1 tablet, and then coated to prevent moisture absorption. Capsule-type Kimchi powder was stored at-20, 0, 4, and 25℃ for 4 months to establish the expiration date of Kimchi powder, then monitored the number of lactic acid bacteria. The number of lactic acid bacteria showed the highest at-20℃, and Kimchi powder stored at 0 and 4℃ also showed over 10⁷ CFU/g. However, lactic acid bacteria was not detected at Kimchi powder stored at 25℃, indicating that Kimchi powder must be distributed under cold-chain system. In case of freezing storage (-20℃), lactic acid bacteria in Kimchi powder can be preserved over 6 months.
D. Application Plan of Results
- In this study, Kimchi powder was developed by setting up the freeze-drying and micro-particle conditions, in addition, capsule, granula, and tablet conditions for commercialization of Kimchi powder were established.
- The study is expected to be published in patents and papers. Also we willplan to conduct to the media relations "Kimchi powder-living lactic acid bacteria".
- Research results can be applied for developing new-product in various fields such as soup-base or confectionery and bakery-base products.

목차 Contents

• 표지 ... 1
• 제출문 ... 2
• 요약문 ... 3
• SUMMARY ... 21
• CONTENTS ... 30
• 목차 ... 32
• 제 1장 연구개발과제의 개요 ... 34
• 제 1절 연구개발의 필요성 ... 34
• 제 2절 연구개발 목표 및 범위 ... 35
• 제 2장 국내외 기술개발 현황 ... 43
• 제 1절 특허분석 측면 ... 43
• 제 2절 논문분석 측면 ... 45
• 제 3절 제품 및 시장분석 측면 ... 48
• 제 3장 연구개발수행 내용 및 결과 ... 50
• 제 1절 [1세부과제]김치발효 미생물 천이조절 우량종균의 개발과 미생물 천이조절 김치의 개발 ... 50
• 제 2절 [2세부과제]수출김치의 산막효모 제어 기술의 개발 ... 144
• 제 3절 [3세부과제]동물실험을 통한 개발제품의 독성 및 기능성 평가 ... 220
• 제 4절 [1협동과제]DGGE기법을 통한 김치 및 원부재료의 미생물 천이 규명 ... 296
• 제 5절 [2협동과제]종균 미생물의 대량 배양 및 안정화 기술 개발 ... 331
• 제 6절 [3협동과제]김치발효 미생물 천이조절 우량종균의 개발과 미생물 천이조절 김치의 개발 ... 365
• 제 4장 목표달성도 및 관련분야에의 기여도 ... 382
• 제 1절 연도별 연구목표 및 평가착안점 ... 382
• 제 2절 연구개발 목표의 달성도 및 관련분야의 기술 발전에의 기여도 ... 383
• 제 5장 연구개발 성과 및 성과활용 계획 ... 397
• 제 1절 실용화․산업화 계획 ... 397
• 제 2절 교육지도․홍보 ... 398
• 제 3절 논문․특허․인력양성 ... 399
• 제 6장 연구개발과정에서 수집한 해외과학기술정보 ... 405
• 제 1절 동물실험을 통한 개발제품의 독성 평가 ... 405
• 제 2절 DGGE기법을 통한 김치 및 원부재료의 미생물 천이 규명 ... 405
• 제 3절 분말김치의 개발과 상품화 ... 406
• 제 7장 연구시설‧장비 현황 ... 408
• 제 8장 참고문헌 ... 409
• 제 1절 [1세부]김치발효 미생물 천이조절 우량 종균의 개발과 미생물 천이조절 김치의 개발 ... 409
• 제 2절 [제2세부]수출김치의 산막효모 제어 기술의 개발 ... 410
• 제 3절 [제3세부]동물실험을 통한 개발제품의 독성 및 기능성 평가 ... 411
• 제 4절 [제1협동]DGGE기법을 통한 김치 및 원부재료의 미생물 천이 규명 ... 412
• 제 5절 [제2협동]종균 미생물의 대량 배양 및 안정화 기술 개발 ... 415
• 제 6절 [제3협동]분말김치의 개발과 상품화 ... 416
• 끝페이지 ... 418