본 시험은 마늘 절임에서 분리 동정된 Leuconostoc citreum SK2556 균주에 의한 마늘 발효 배양액 급여가 산란계의 계란 생산성, 계란 품질, 혈액 성상에 미치는 영향을 알아보고자 실시하였다. 시험 동물은 33주령 산란계(Hy-Line Brown) 180수를 공시하였고, 5주간 사양시험을 실시하였 다. 전체 사양기간동안 산란율은 대조구, FG (발효마늘) 0.3 및 FG 0.5 처리구는 78.02~78.53의 범위로 유의적인 차이가 없었으나, FG 0.1 처리구(75.37±3.22)는 다른 처리구들에 비교하여 감소하였다(P<0.05). 사료섭취량과 사료요구율에서는 유의한 차이가 관찰되지 않았다. 시험 전기간 Haugh unit에서는 마늘 발효액 FG 0.1이 대조구와 FG 0.5에 비해 유의하게 증가하였다(P<0.05). 혈액 내 Glucose는 모든 마늘 발효액 처리구가 대조구보다 유의하게 낮았다(P<0.01). 간 기능의 이상 여부를 나타내는 알칼리인산분해효소(alkaline phosphatase; ALP)는 FG 0.3 처리구가 대조구보다 유의하게 감소하였다(P<0.05). 본 연구 결과, 산란계 사료 내 마늘 발효 배양액 급여는 계란의 Haugh unit의 향상으로 계란의 신선도를 개선시켰을 뿐만 아니라, 건강 유지에 긍정적인 영향을 미쳐 사료첨가제로서의 이용가능성을 제시하고자 한다.

Leuconostoc spp. are generally utilized as kimchi starters, because these strains are expected to have beneficial effects on kimchi fermentation, including improvement of sensory characteristics. Here, we developed a detection method for verifying the presence of the kimchi starter Leuconostoc mesenteroides WiKim33, which is used for control of kimchi fermentation. A primer set for multiplex polymerase chain reaction was designed based on the nucleotide sequence of the plasmids in strain WiKim33, and their specificity was validated against 45 different strains of Leuconostoc spp. and 30 other strains. Furthermore, the starter strain consistently tested positive, regardless of the presence of other bacterial species in starter kimchi during the fermentation period. Our findings showed that application of a strain-specific primer set for strain WiKim33 presented a rapid, sensitive, and specific method for detection of this kimchi starter strain during natural kimchi fermentation.

여주(Momordica charantia) 열수 추출물을 젖산균(Leuconostoc mesenteroidesies)으로 발효하여 얻은 생산물의 항산화 및 항당뇨 효과를 관찰하였다. 여주 발효물(MC-LM)과 여주 추출물(MC)의 항산화 활성은 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)(ABTS) radical 소거 측정법을 사용하여 관찰하였다. MC 처리군은 radical 소거활성이 거의 나타나지 않았으나 MC-LM의 경우 500 μg/mL 농도에서 47%의 활성이 관찰되었으며 MC-LM의 농도가 증가함에 따라 활성이 유의하게 (p<0.05) 증가하는 것을 확인 할 수 있었다. MC-LM은 MC에 비해 peroxyl radical에 의한 DNA의 산화적 손상을 더 효과적으로 억제하였으며 MC-LM의 농도가 증가함에 따라 strand breakage의 억제가 유의적으로 증가 하였다(p<0.05). 혈당량과 밀접한 관련이 있는 α-glucosidase의 활성 억제정도를 측정한 결과 MC는 8 mg/mL 농도에서 α-glucosidase의 활성을 30.1% 감소시켰고 같은 농도에서 MC-LM은 58.9% 감소 시켰다. 또한 alloxan에 의한 RIN-m5F 췌장세포 사멸에 발효물의 영향을 관찰한 결과 MC-LM을 농도별로 처리한 군에서 세포생존율이 유의적으로(p<0.05) 증가하였으며 1000 μg/mL 농도에서 MC군 보다 20% 높게 증가되었다. Alloxan에 의한 β-cell파괴를 유도하여 RIN-m5F cell이 분비한 인슐린의 측정결과 MC-LM을 농도별로 첨가한 모든 군에서 인슐린 분비능이 증가하는 경향을 보였으며, 1000 μg/mL 농도에서 MC군 보다 15% 높게 증가되었다. 결론적으로 본 연구에서 관찰된 결과들을 통해 여주 발효물은 항산화 및 항당뇨 효과가 있는 것으로 확인되었다.

본 연구에서는 김치로부터 L. mesenteroides CJNU0041 을 분리하여 16S rRNA 염기서열 분석을 통해, 동정하였 으며, β-glucosidase 활성이 우수한 것으로 나타났다. L. mesenteroides CJNU0041를 이용한 브로콜리 발효 동안 생 균수와 pH 및 β-glucosidase 활성에 대해 분석한 결과, 발효 시간은 48 hours이 적당한 것으로 판단되었다. 또한, 발효 후 생물 전환이 일어남을 HPLC 분석을 통해 확인 하였다. 그리고, L. mesenteroides CJNU0041의 브로콜리 발효물에서 C. difficile에 대한 생육 억제 효과를 확인 할 수 있었다. 따라서, 본 연구를 통해 분리된 신규 L. mesenteroides CJNU0041을 C. difficile의 생육 제어를 위한 유산균 제재화가 가능할 것으로 판단되며, 브로콜리 발효물도 다양한 식품에 적용이 가능할 것으로 사료된다.

Glucansucrase is an enzyme classified as a glycoside hydrolase (GH) 70 family, which catalyzes the synthesis of glucooligosaccharides with a low molecular weight using sucrose as a donor of D-glucopyranose and maltose as a carbohydrate acceptor. In this study, glucansucrase-producing lactic acid bacteria strain was isolated from the fermented foods collected in traditional markets, and the optimum conditions for the oligosaccharide production were investigated. The strain CCK940 isolated from Chinese cabbage kimchi was selected as an oligosaccharide-producing strain due to its high glucansucrase activity, with 918.2 mU/mL, and identified as Leuconostoc lactis. The optimum conditions for the production of oligosaccharides using Leu. lactis CCK940 were to adjust the initial pH to 6.0, add 5% (w/v) sucrose and 10% (w/v) maltose as a donor and acceptor molecules, respectively, and feed 5% (w/v) sucrose at 4 and 8 h of cultivation. When Leu. lactis CCK940 was cultured for 12 h at optimum conditions, at least four oligosaccharides with a polymerization degree of 2-4 were produced.

Glucansucrase is an enzyme classified as a glycoside hydrolase (GH) 70 family, which catalyzes the synthesis of various glucans and glucooligosaccharides with a low molecular weight using sucrose as a donor of D-glucopyranose and maltose as a carbohydrate acceptor. Oligosaccharides are indigestiable carbohydrate with low calories, which have various positive effects on the health of intestinal track. In this study, oligosaccharide produced from Leuconostoc lactis CCK940 was purified and its prebiotics effect was investigated. Leuconostoc lactis CCK940 which isolated from Chinese cabbage kimchi produced oligosaccharide using 5%(w/v) sucrose as a donor and 3%(w/v) maltose as an acceptor. Oligosaccharide produced was purified using Bio-gel P2 column chromatography and DPs of the resulting oligosaccharide were ranged from 3 to 7. When the prebiotics effects of freeze dried oligosaccharide were examined, 1%(w/v) of purified oligosaccharide added to carbohydrate-free MRS showed good prebiotics effects on Lactobacillus plantarum KCCM 12116, L. reuteri KCTC40417, and Bifidobacterium animalis KCCM1209.

For preliminary molecular typing, PCR-based fingerprinting using random amplified polymorphic DNA (RAPD) is the method of choice. In this study, 14 bacterial strains were isolated from different Korean food sources, identified using 16S rRNA gene sequencing, and characterized through RAPD-PCR. Two PCR primers (239 and KAY3) generated a total of 130 RAPD bands, 14 distinct PCR profiles, 10 polymorphic bands, one monomorphic band, and four unique bands. Dendrogram-based analysis with primer 239 showed that all 14 strains could be divided into seven clades out of which clade VII had the maximum of seven. In contrast, dendrogram analysis with the primer KAY3 divided the 14 L. citreum strains into four clades out of which clade IV consisted of a maximum of 10 strains out of 14. This research identified and characterized bacterial populations associated with different Korean foods. The proposed RAPD-PCR method, based on sequence amplification, could easily identify and discriminate the lactic acid bacteria species at the strain-specific level and could be used as a highly reliable genomic fingerprinting tool.

For preliminary molecular typing, PCR-based fingerprinting using random amplified polymorphic DNA (RAPD) is the method of choice. In this study, 14 bacterial strains were isolated from different Korean food sources, identified using 16S rRNA gene sequencing, and characterized through RAPD-PCR. Two PCR primers (239 and KAY3) generated a total of 130 RAPD bands, 14 distinct PCR profiles, 10 polymorphic bands, one monomorphic band, and four unique bands. Dendrogram-based analysis with primer 239 showed that all 14 strains could be divided into seven clades out of which clade VII had the maximum of seven. In contrast, dendrogram analysis with the primer KAY3 divided the 14 L. citreum strains into four clades out of which clade IV consisted of a maximum of 10 strains out of 14. This research identified and characterized bacterial populations associated with different Korean foods. The proposed RAPD-PCR method, based on sequence amplification, could easily identify and discriminate the lactic acid bacteria species at the strain-specific level and could be used as a highly reliable genomic fingerprinting tool.

Dextran is a generic term for a bacterial exopolysaccharide synthesized from sucrose and composed of chains of D-glucose units connected by α-1,6-linkages by using dextransucrases. Dextran could be used as vicosifying, stabilizing, emulsifying, gelling, bulking, dietary fiber, prebiotics, and water holding agents. We isolated new strain capable of producing dextran from Korean traditional kimchi and identified as Leuconostoc sp. strain JYY4. Batch fermentation was conducted in bioreactor with a working volume of 3 L. The media was MMY and 15% (w/v) sucrose. Mineral medium consisted of 3.0 g KH2PO4, 0.01 g FeSO4, H2O, 0.01 g MnSO4, 4H2O, 0.2 g MgSO4 7H2O, 0.01 g NaCl, 0.05 g CaCl2 per 1 liter deionized water. The pH of media was initially adjusted to 6.0. The inoculation rate was 1.0% (v/v) of the working volume. Temperature was maintained at 28oC. The agitation rate was 100 rpm. The production pattern of dextran was associated with the cell growth. After 24 hr dextran reached its highest concentration of 59.4 g/L. The sucrose was consumed completely after 40 hr. Growth reached stationery phase when sucrose became limiting, regardless of the presence of fructose or mannitol. When the specific growth rate was 0.54 hr-1, utilization averaged 5.8 g/L-hr. The yield and productivity of dextran were 80% and 2.0 g/L-hr, respectively. Dextrans produced by were separated to two different size by an alcohol fraction method. The size of high molecular weight dextran (45% alcohol, v/v), less soluble dextran, was between MW 500,000 and 2,000,000. Soluble dextran (55% alcohol, v/v) was between 70,000 and 150,000. The molecular weight average of total dextran (70% alcohol, v/v) was between 150,000 to 500,000. The enzymatic hydrolyzates of total dextran of ATCC 13146 showed branched dextrans by Penicillium dextranase contained of glucose, isomaltose, isomaltotriose, and isomaltooligosaccharides greater than DP4 (degree of polymerization) that had branch points. Compounds greater than DP4 were branched isomaltooligosaacharides. Hydrolysates by the Lipomyces dextranase produced the same composition of oligosaccharides as those by Penicillin dextranase.

Lactic acid bacteria (LAB) are known to promote activation of macrophages, selectively eliminate harmful intestinal bacteria that enter the body, and increase secretion of cytokines, chemokines, and inflammatory mediators that boost the immune system. In the current study, the immune-enhancing effects of five Leuconostoc stains (Leu. kimchii WK18, Leu. citreum DH33, Leu. mesenteroides DH34, SE34, and GY17) isolated from traditional Korean fermented food, Kimchi, were evaluated. Lactobacillus rhamnosus GG (LGG), which is known for its extraordinary immunopotentiating activity, was used as a reference strain. According to the results, several heat-killed Leuconostoc strains induced a higher level of NO, IL-1β, or TNF-α production than the LGG strain. Collectively, results of the current study suggest that Leuconostoc strains could enhance immune response through activation of macrophages, and could be significant references when verifying the possibility for use of Leuconostoc strains as probiotics, starter, dietary supplement, or immune-enhancing medicine.

효소반응과 부분 산가수분해 결과를 해석하여 Leuconostoc mesenteriodes M-12 덱스트란수크라제 억셉터 반응 산물이 새로운 분지올리고당의 구조를 확인하였다. 분지올리고당 B_4의 구조는6^2-O-α-D-kojibiosylmaltose인 것으로 확인되었으며, 분지올리고당 B_5의 구조는 6^3-O-α-D-kojibiosylpanose였다. 억셉터 반응산물을 덱스트라나제로 분해한 결과 새로운 올리고당인 D_4를 확인할 수 있었다. 억셉터 반응의 산물을 억셉터로 이용한 두 번째 억셉터 반응의 생성물을 덱스트라나제 처리하여 D_4를 얻었는데 덱스트라나제와 글루코아밀라제에 의해 분해되지 않았다. 그 구조는 6^2-O-α-D-kojibiosylisomaltose로 확인되었다. 직선상 또는 분지결합을 가진 d.p. 6이하의 억셉터 반응산물의 생성 패턴도 확인하였다.

Grapefruit extract rich in functional substances adjusting pH 5.0 and adding sucrose (final concentration, 2%) was fermented by Leuconostoc mesenteroides CJNU 0147 strain, consequently prepared a fermented grapefruit extract containing dextran. As a result of analyzing the growth inhibitory effect on gut harmful bacterium Clostridium difficile strain using the prepared fermented grapefruit extract, the viable cell count was significantly reduced (p<0.05 vs. control for 0.5 brix as a final concentration; p<0.001 vs. control for 1.0 brix). On the other hand, the proliferative ability of the extract for Bifidobacterium breve strain, which is one of the species of the genus Bifidobacterium well known as human gut beneficial bacteria, was confirmed (p<0.001 vs. control for both 0.5 and 1.0 brix). These results indicate fermented grapefruit extract with Leu. mesenteroides CJNU 0147 strain inhibits the growth of gut harmful bacterium C. difficile and promotes the proliferation of beneficial bacterium B. breve and is expected to be used as a functional food material for gut health.

Sweet pumpkin paste (SPP) was fermented by Leuconostoc mesenteroides SM at 25℃ for 3 days for enhancing its physicochemical properties. SPPs with 5%, 10%, and 15% solid contents (SC) were fortified with 20% sucrose and 0.5% yeast extract. The unfermented SPP with 15% SC indicated L, a, and b color values of 25.02, 4.66, and 13.35, respectively, and a consistency index of 48.6 Pa·sn. During the 3 days of fermentation, both the a and b color values decreased slightly, whereas the consistency index increased to 188.8 Pa·sn, giving the fermented product a pudding-like consistency. This fermented SPP (15% SC) showed the highest acid production and viable cell counts among samples, indicating pH 3.85, 1.30% acidity and 9.2×108 CFU/mL respectively. The added sucrose was completely utilized after 1 day of fermentation. After 3 days, the insoluble and soluble dextran contents were 8.9% and 4.5%, respectively. Furthermore, the contents of mannitol and fructose were 3.11% and 1.76%, respectively. Regarding the sensory evaluation, this fermented sample also indicated the highest color, taste and texture scores, and was the overall preferred sample. In conclusion, the fermented SPP with 15% SC was carotinoid-rich a wholesome pumpkin-based product that is rich in probiotics and lactic bacteria-produced mannitol and dextran, which gave the product an acceptable viscous pudding-like consistency and good organoleptic properties.

Many traditional fermented foods contain diverse bacteria and many lactic acid bacteria (LAB) play important roles for the fermentation of foods such as many dairy foods and kimchi. Especially, Leuconostoc mesenteroides is one of major bacteria in those fermented foods and the development of this species would be expected to be critical for strain improvement as well as the industrialization. Up to now, a lot of plasmids were isolated from Leuconostocs species including Leu. mesenteroides and a number of vector systems has been developed. Many plasmid vector systems using Leuconostocs spp. employ RCR (rolling circle replication) producing single-stranded DNA intermediates or sometimes takes theta replication. These plasmids include the sequences for Pre protein, recombination specific sites such as RSA (recombination site A) and RSB (recombination site B), and single strain origins for RCR replication. This information might be helpful to elucidate the improvement of Leuconostocs spp. and moreover the development of diverse products using Leuconostocs spp.

Antioxidant activity is important for reducing oxidative stress that causes various metabolic disorders. Metabolic disorders are highly related to loss of productivity in livestock. Therefore, development of effective antioxidant compounds originating from plants is important for organic agriculture. Phenolic compounds in edible plants are regarded as major components relevant to antioxidant activity. The present study investigated the changes in antioxidant activity and phenolic compound profiles of Aronia (Aronia meloncarpa) by fermentation using different strains of Leuconostoc mesenteroides. A total of 5 strains of L. mesenteroides were used as starter cultures and their β-glucosidase activities were measured. A total of 6 experiment runs were prepared, one for control (uninoculated) and the others (inoculated) for treatments. For biological activity, antioxidant and antibacterial activities were measured. For phenolic compound profiling, TLC and HPLC analysis were performed. The strains of KACC12313 and KACC12315 showed greater enzyme activity than others. Treatment with KCCM35046 showed strong and broad antibacterial activity against to Listeria monocytogenes. Treatments with KCCM35046 and KACC12315 showed the highest total polyphenol content. The highest antioxidant activity was found in KACC12315 treatment. No remarkable alteration was found in thin layer chromatography (TLC) analysis. In phenolic compound profiling analysis, KCCM35046 showed notable alteration in compound area ratio compared to others and also showed the highest caffeic acid content. In chlorogenic acid, treatments with KCCM35046 and KACC12315 showed great content than others. Treatment with KACC12315 showed the greatest content of trans-ferulic acid. As a result of relative performance indexing analysis, L. mesenteroides KCCM35046 and KACC12315 were selected as the best strain for the fermentation of Aronia.

The present study was carried out to investigate the in vivo single-dose acute toxicity of Leuconostoc citreum GR1 (Leuc. citreum GR1), a lactic acid bacteria isolated from kimchi, in ICR male and female mice. The test article was orally administered once to both sexes of mice. The mortalities, clinical findings, autopsy findings, and body weight changes were monitored daily for two weeks. The male and female mice were gavaged with Leuc. kimchi GR1 of four doses (625, 1,250. 2,500 and 5,000 mg/kg). The oral LD50 of Leuc. citreum GR1 was considered higher than 5,000 mg/kg. No significant changes in the general conditions, body weights, clinical signs and presence of gross lesions were observed in both sexes of mice to whom Leuc. citreum GR1 was administered orally. The results indicated that the 5,000 mg/kg dose of Leuc. citreum GR1 showed no adverse effect.

자연적으로 일어나는 형질전환은 세균에서 유전 요소의 수평적 전달 메커니즘의 하나로서 세균 생리학에서 빠질 수 없을 만큼 중요한 현상이다. S. thermophilus는 자연적으로 형질전환이 가능한 상태로 변환하기 위해서 Rgg-타입의 response regulator인 ComR과 소수성 펩타이드 페르몬인 ComS를 포함하는 새로운 quorum sencing 시스템을 이용한다. S. thermophilus를 화학적 제한배지에서 배양하면 대수기 초기에 신호서열이 분리된 ComS는 세포외부로 배출된다. 배출된 ComS는 올리고 펩타이드 전달 시스템인 Ami를 통하여 세포내로 유입되어 ComR과 결합하게 된다. ComS와 결합된 활성화된 ComR은 후기 형질전환 관련 유전자들의 전사를 조절하는 대체형 시그마펙터(σ^x)를 암호화하는 comX 유전자의 발현을 유도함으로써 형질전환이 일어나도록 만든다. 신호서열이 제거된 ComS를 합성하여 화학적 합성배지에 첨가하게 되면, S. thermophilus의 형질전환을 인공적으로 유도할 수 있다. Leuconostoc 속 유산균들 은 그들이 후기 형질전환 관련 유전자들과 서열이 유사한 단백질 유전자들을 보유하고 있음에도 불구하고 자연적 형질전환 메커니즘은 아직 밝혀지지 않았다. Leuconostoc 속의 자연적 형질전환의 메커니즘을 규명하는 연구는 김치산업에서 사용될 수 있는 개량된 균주 개발을 가능하게 할 것이다.