Human malignant melanoma is an aggressive skin cancer which has been rising at a greater rate than any other cancers. Although various new therapeutic methods have been developed in previous studies, this disease has properties of high proliferation and metastasis rate which remain obstacles that have lead to a poor prognosis in patients. It has been reported that a specific Lactobacillus extract has anti-cancer and –metastasis effect in vitro and in vivo. However, previous research has not specified precisely what effect the Lactobacillus rhamnosus GG (LGG) extract has had on human malignant melanomas. In this study, we showed that the LGG extract has anti-cancer and –metastasis effects on the human malignant melanoma cell lines, A375P and A375SM. At first, it was found that, while the LGG extract affects human neonatal dermal fibroblasts slightly, it induced the dose-dependent anti-cancer effect on A375P and A375SM by a WST-1 proliferation assay. As a result of a real-time PCR analysis, the expression patterns of several genes related to cell cycle, proliferation, and apoptosis were modulating in a manner that inhibited the growth of both malignant melanoma cell lines after the treatment of the LGG extract. Furthermore, genes related to the epithelialmesenchymal transition were down-regulated, and migration rates were also decreased significantly by the LGG extract. Our study showed that the LGG extract could be used as a potential therapeutic source.

Alginate lyase from Streptomyces violaceoruber was purified by DEAE sephacel chromatography and SP sepharose chromatography. The specific activity of the purified enzyme was 14.6 units/mg protein, representing a 40.6-fold purification of the crude extract. The final preparation thus obtained showed a single band on Tricine-SDS polyacrylamide gel electrophoresis whose molecular weight was determined to be 23.3 kDa. The polyMG block of sodium alginate was hydrolyzed by the purified alginate lyase and then separated by activated carbon column chromatography and bio gel P-2 gel filtration. The main hydrolysates were composed of hetero type M/G-oligosaccharides with the degrees of polymerization (D.P.) being 6 and 8. To investigate the effects of hetero type M/Goligosaccharides from the sodium alginate on the growth of some intestinal bacteria, cells were cultivated individually on the modified-MRS medium containing D.P. 6 and 8 M/G-oligosaccharides. B. longumgrew 4.25-fold and 6.44-fold more effectively by the treatment of D.P. 6 and 8 M/G-oligosaccharides compared with those of standard MRS medium. In addition, B. bifidumgrew 3.3-fold and 5.4-fold more effectively by the treatment of D.P. 6 and 8 M/G-oligosaccharides. In conclusion, D.P. 8 was more effective than D.P. 6 hetero M/G-oligosaccharides as regards the growth of Bifidobacteriumspp. and Lactobacillus spp

Alginate lyase from Streptomyces violaceoruber was purified by DEAE sephacel chromatography and SP sepharose chromatography. The specific activity of the purified enzyme was 14.6 units/mg protein, representing a 40.6-fold purification of the crude extract. The final preparation thus obtained showed a single band on Tricine-SDS polyacrylamide gel electrophoresis whose molecular weight was determined to be 23.3 kDa. The polyMG block of sodium alginate was hydrolyzed by the purified alginate lyase and then separated by activated carbon column chromatography and bio gel P-2 gel filtration. The main hydrolysates were composed of hetero type M/G-oligosaccharides with the degrees of polymerization (D.P.) being 6 and 8. To investigate the effects of hetero type M/Goligosaccharides from the sodium alginate on the growth of some intestinal bacteria, cells were cultivated individually on the modified-MRS medium containing D.P. 6 and 8 M/G-oligosaccharides. B. longumgrew 4.25-fold and 6.44-fold more effectively by the treatment of D.P. 6 and 8 M/G-oligosaccharides compared with those of standard MRS medium. In addition, B. bifidumgrew 3.3-fold and 5.4-fold more effectively by the treatment of D.P. 6 and 8 M/G-oligosaccharides. In conclusion, D.P. 8 was more effective than D.P. 6 hetero M/G-oligosaccharides as regards the growth of Bifidobacteriumspp. and Lactobacillus spp. Key words: hetero M/G-oligosaccharides, Streptomyces violaceoruber

Fermented food consists of a variety of lactic acid bacteria (LAB) that are also used in the industry as starter cultures. This genus comprises of different species that find importance as a preservatives in food like meat and sausages. Likewise, Lactobacillus brevis has been recognized as GRAS and is a potential probiotic strain extensively being used on an industrial scale. Since such bacteria are directly related to human health, there has been a need to identify and characterize them on the molecular level. In this study, LAB was identified and characterized from various fermented food samples available in South Korea. Two types to PCR-based molecular typing methods were used to analyze the 13 Lactobacillus brevis isolates, of which one was based on difference in the banding patterns originated on agarose gel and the other was related to the sequence analyses of various housekeeping genes in the particular strain. The former rep-PCR technique used three primers namely, REP, ERIC and (GTG)5 that amplified repetitive sequences in the genome and provided characteristic fingerprint profile for each isolate. This clustered the strains in 3 groups with the help of UPGMA method of clustering distinguishing between closely related strains. However, the latter multilocus sequencing typing (MLST) technique provided definite identification of the strains. A set of 7 housekeeping genes were determined as groEL, gyrB, rpoA, rpoB, pheS, recA and dnaK. These genes were amplified and sequenced and were subjected to comparative analysis. Discrete allelic profiles and 13 sequence types (STs) were resolved and minimum spanning tree (MST) was constructed, revealing the genetic relatedness among the isolates. On comparing the results from both the techniques, MLST proved to generate accurate and precise fingerprints owing to the sequence analysis of conserved genes thus providing a scope for research in the monitoring related species.

이 연구는 고지방 및 고 콜레스테롤식이(HD)를 먹인 rat에서의 저지질혈증 및 항염증 효과에 영향을 미치는 카제인과 유당(cMRP) 그리고 락토바실러스 fermentum H9 (FcMRP)에 의해 발효된 cMRP에 의해 반응된 마일라드 반응 생성물의 효과를 조사했다. HD 식이를 실시한 rat은 정상적인 식이를 먹인 rat과 비교했을 때, 간의 트리글리세리드 농도가 유의적으로 증가했다. 이는 HD 그룹에 비해서 Simvastatin, L. fermentum H9(H9), cMRP, FcMRP의 처리기간의 총 트리글리세리드를 감소시킨 것으로 나타났다. 조직학적 분석에서 cMRP, F-cMRP 및 H9-를 먹인 rat에서 간과 대동맥 조직의 지질 축적이 감소한 것으로 관찰되었다. 또한, F-cMRP와 cMRP는 HD 그룹에서 내중막 두께를 감소시켰다. 게다가 H9, Cmrp, F-cMRP 처리는 ICAM-1 및 VCAM-1의 발현수준을 유의적으로 감소시켰지만, MCP-1의 발현 수준은 유의하게 감소시키지 못했다. 특히, ICAM-1과 VCAM-1의 발현은 HD 그룹에 비해 F-cMRP 그룹에서 유의적으로 감소했다. 이 연구 결과는 낙농 식품의 cMRP와 F-cMRP가 심혈관계 질환, 특히 죽상 동맥 경화증을 예방하거나 치료하는데 잠재적으로 사용될 수 있음을 시사한다.

In this study, we found that the glycoprotein (18 kDa) isolated from Lactobacillus plantarum L67 (L67 protein) inhibits allergy-related inflammatory mediators in BPA treated rat basophilic leukemia 2H3 (RBL-2H3) cells and primary cultured splenocytes. First, we measured the levels of ERK, p38, and AP-1 by Western blotting and we evaluated the expression of IL-1β, IL-6, and IL-10 cytokines by real-time quantitative PCR. Finally, we evaluated the applicability of L. plantarum L67 as a yogurt starter culture by measuring pH, enumeration of bacteria, and sensory scores. The results of this study shows that L67 protein inhibits the phosphorylation of ERK and p38 MAPK, as well as the activation of AP-1 (c-Fos and c-Jun). With inflammationrelated factors (Th2-related cytokines), L67 protein inhibits BPA-induced IL-1β, IL-6, and IL-10 activity. Bisphenol A stimulates MAPK signaling, which induces increased GATA-3 expression, and GATA-3 induces the differentiation of Th2 cells and secretion of Th2-related cytokines. Our findings showed that L67 protein may regulate the activation of GATA3 and T-bet through the induction of MAPK signaling, and it relieves the Th1/Th2 cytokine profile imbalance. Taken together, these findings demonstrate that the L. plantarum L67 strain is a potential food supplement and starter culture, that could help in the prevention of Th2-related immune disorders.

Lactic acid bacteria (LAB) producing phenyllactic acid (PLA), which is known as antimicrobial compound, was isolated from button mushroom bed and the isolated LAB was identified to Lactobacillus casei by 16 rRNA gene sequence analysis. Cell-free supernatant (CFS) from L. casei was assessed for both the capability to produce the antimicrobial compound PLA and the antifungal activity against three fungal pathogens (Rhizoctonia solani, Botrytis cinerea, and Collectotricum aculatum). PLA concentration was investigated to be 3.23 mM in CFS when L. casei was grown in MRS broth containing 5 mM phenylpyruvic acid as precursor for 16 h. Antifungal activity demonstrated that all fungal pathogens were sensitive to 5% CFS (v/v) of L. casei with average growth inhibitions ranging from 34.58% to 65.15% (p < 0.005), in which R. solani was the most sensitive to 65.15% and followed by C. aculatum, and B. cinerea. The minimum inhibitory concentration (MIC) for commercial PLA was also investigated to show the same trend in the range of 0.35 mg mL-1 (2.11 mM) to 0.7 mg mL-1 (4.21 mM) at pH 4.0. The inhibition ability of CFS against the pathogens were not affected by the heating or protease treatment. However, pH modification in CFS to 6.5 resulted in an extreme reduction in their antifungal activity. These results may indicate that antifungal activities in CFS was caused by acidic compounds like PLA or organic acids rather than protein or peptide molecules.

For several decades, lactic acid bacterium (Lactobacillus graminis: LAB) has been generally recognized as safe. To develop the pan-environmental bio-control agent, algicidal activity of the live LAB cell and its culture filtrate (CF) was examined against Microcystis aeruginosa. LAB cells perfectly lysed M. aeruginosa within 3 days, while the CF had a less effect than the live cells, approximately 78% inhibition of algal growth during a same culture period. The concentration of microcystin in alone culture of M. aeruginosa was 7.1 μg L-1, but gradually increased and leach 158.5 μg L-1 on 10 days. However, LAB cells clearly decreased the microcystin by 10.3 μg L-1 in the same period, approximately 93.5%. CF of LAB showed a strong algicidal activity over 75% between pH 2-7, 91.3% by the treatment of proteinase K, 87.8% by below 3 kDa in particle size, and 75.3% by heat treatment, respectively. Of five solvents, fractions of CF passed through solvents diethyl ether and ethyl acetate showed an obvious algicidal activity in the algal-lawn test. Among 5 fractions purified by silica-gel TLC plate, two spots showed a most strong removal activity on M. aeruginosa. Another analysis of GC indicate that CF contained six representative fatty acids. Even though most of these substance have been known as an anti-algal substance against M. aeruginosa, oleic acid is the most effective. These results suggested that the culture filtrate or specific substances, like a fatty acids, in comparison with live L. graminis can be a successful and eco-friendly agent to control Microcystis bloom.

α-Amylase 및 protease 활성을 동시에 가지는 1-B-12 균주를 된장으로부터 분리하였다. 16S rRNA 유전자 염기서열 분석결과, Bacillus licheniformis로 동정되어 B. licheniformis 1-B-12 균주로 명명하였다. 효소활성 측면에서 현미(50%), 미강(30%) 및 대두(20%)의 조합이 최적의 기질 조합 조건으로 확인되었다. 기능성이 부가된 효소식품을 제조하기 위하여 Lactobacillus casei GW140 균주를 도입하였는데, B. licheniformis 1-B-12 균주와의 혼합배양 조건은 2단발효(B. licheniformis 1-B-12 접종 > 24 h 발효 > L. casei GW140 접종 > 12 h 추가 발효)가 적합한 것으로 나타났다. 이러한 최적 조건으로 생산된 효소식품을 동결건조하여 25℃에서 보관하였을 때 시험 마지막 날인 45일까지 효소의 활성이 유지되었다.

비피도박테리아 증식인자인 1,4-dihydroxy-2-naphthoica cid 생산 유산균주 LP2가 선행연구에서 자연치즈로부터 분리되었다. LP2 균주는16S rRNA 유전자 염기서열 분석에 의한 균주동정에서 Lactobacillus plantarum(99% 일치율)과 가장 높은 상동성을 나타내어 Lactobacillus plantarum LP2 균주로 명명하였다. LP2 균주의 배양 상등액은 Helicobacter pylori KCTC 12083 균주에 대하여 항균활성을 나타내었으며 이는 생산된 DHNA에 기인하는 것으로 추측되었다.

본 연구에서는 호밀을 이용하여 사일리지 제조시 Lactobacillus plantarum KCC-10 및 KCC-19를 접종하여 사일리지의 사 료가치, 품질 및 미생물상의 변화를 조사하였다. 호밀 사일 리지에서 젖산균 접종구와 무접종구에서 사료가치는 크게 변화되지 않았다. 그리고 유산 함량은 젖산균 접종구가 현 저하게 높은 경향을 보였으나 초산과 낙산은 젖산균 접종 구가 약간 낮아지는 경향을 보였다. 젖산균 분포는 젖산균 접종구에서 현저하게 높았으며 효모와 곰팡이는 104CFU/g 에서 관찰되지 않았다. 호밀 사일리지 제조시 KCC-10 및 KCC-19 접종구의 조단백질 및 TDN 함량은 무접종구 보 다 약간 증가하였으나 섬유소 및 in vitro 건물소화율에서 는 비슷한 수준을 보였다. 이상의 결과를 요약해 보면 호 밀 사일리지 제조시 KCC-10 및 KCC-19 접종에 의해 사일 리지의 품질이 향상되었다.

This study was to investigate whether the bioactivity of L. plantarum L67 can be enhanced by treatment with Zanthoxylum piperitum DC (ZPDC) glycoprotein or compounds of similar molecular weights (24 kDa). When L. plantarum L67 was treated with ZPDC glycoprotein of various concentrations, the intensities of a few SDS-PAGE bands were slightly changed. The amount of a 23-kDa protein was increased upon treatment with increasing concentrations of ZPDC glycoprotein. The activities of antioxidant enzymes (SOD, GPx, and CAT), growth rate, and β-galactosidase activity were more increased at treatment with ZPDC glycoprotein than at treatment of L. plantarum L67 only, whereas the ATPase activity was decreased. Interestingly, it did not affect scavenging activity for the DPPH. The founding result suggest that, one of beneficial strategies for increase the bioactivity of L. plantarum L67 might be to take ZPDC.