To increase industrial applicability of Astragalus membranaceus (AM) as immunostimulating materials, hot-water extract (AME) was prepared from AM and fermented with Kimchi-lactic acid bacteria (Lactobacillus sakei & Leuconostoc mesenteroides) to prepare fermented AM-postbiotics (FAME). Although FAME prepared from AM-postbiotics did not show a significant enhancement in macrophage stimulating activity compared to non-fermented AME, crude polysaccharide (FAME-CP) fractionated by EtOH precipitation from FAME showed significantly higher macrophage stimulating activity than AME-CP. Compared to AME-CP, FAME-CP showed dramatic changes in component sugar and molecular weight distribution. FAME-CP was a polysaccharide with a major molecular weight distribution of 113.4 kDa containing Man (44.2%), Glc (19.3%), Gal (10.2%), GalA (10.2%), and Ara (7.4%) as sugar components. FAME-CP with enhanced macrophage stimulatory activity not only increased expression levels of mRNA genes encoding macrophage-activated factors (iNOS, TNF-α, MCP-1, IL-6, and COX-2), but also led the nuclear translocation of activated p65 and c-Jun. In conclusion, crude polysaccharide from AM-postbiotics fermented with lactic acid bacteria could increase industrial applicability as a functional material with enhanced immunostimulating activity than AME-CP.

곽향은 서양에서 Korea mint라고 불릴 정도로 허브와 쓰임새가 매우 유사하다. 서양의 허브는 화장품의 소재로 많이 사용하지만 곽향은 우리나라에서 약재로 더 많이 사용되어 화장품 소재로서의 기능 성을 비교해보았다. 곽향 추출물과 추출물을 이용한 유산균과 효모 발효물, 에센셜오일, 플로럴워터에 대한 항산화, 미백, 폴리페놀, 플라보노이드, 항균력 실험을 진행하였다. 항산화 활성은 모든 샘플에서 높은 효과 가 보였고, 미백과 폴리페놀, 항균력에서 에센셜오일이 가장 높은 효과가 보였다. 반면에 플라보노이드는 70% 에탄올 추출물에서 가장 높은 효과가 보였다. 이런 결과로 에센셜오일이 기능적으로 가장 큰 효과가 있었다. 하지만 추출물을 가지고 효모 발효를 진행한 시료들이 에센셜오일 만큼의 효과가 있다는 것을 확 인해 에센셜오일의 적은 추출량에 비해 효모 발효를 진행한 시료가 화장품소재로서 충분한 가치가 있다고 본다.

Dental caries is an infectious disease that is caused by the interaction between the host, the diet, and a diverse microbial community. This study illustrated the anticariogenic effects of THYJ-15 against Streptococcus mutans, a major agent of dental caries. THYJ-15 was identified as Latilactobacillus sakei subsp. sakei through 16S rRNA sequencing, with a similarity of 99.72% to L. sakei subsp. sakei JCM 1157T. The cell-free supernatant of L. sakei subsp. sakei THYJ-15 observed MIC values of 10 mg/mL and MBC values of 20 mg/mL against S. mutans KACC 16833T. The cell-free supernatant of L. sakei subsp. sakei THYJ-15 showed significant inhibitory effects on biofilm formation, acid production and glucosyltransferases(GTFs) production against S. mutans. As a results, it was confirmed that L. sakei subsp. sakei THYJ-15 is a lactic acid bacterium possessing notable anticariogenic effects. Accordingly, L. sakei subsp. sakei THYJ-15 is expected to have great potential to be used as a basic substance for dental caries treatment caused by S. mutans.

프로바이오틱스는 다양한 환경에서 분리되어 왔으며, Bacillus 균주는 유산균과 같은 일반적인 프로바이오틱스의 한계를 보완할 수 있는 내생포자 형성 능력으로 인해 프로바이오틱스 균주 중에서 유리하다. 본 연구의 목적은 한국에서 오염되지 않은 환경으로 알려진 울릉도 토양에서 분리된 Bacillus 균주의 프로바이오틱스 잠재성을 조사하는 것이다. 토양 시료는 울릉도 각지에 서 채취하였으며, 항생제 내성과 효소 활성을 평가하였다. 항생제에 내성을 나타내지 않는 6개 의 Bacillus 균주를 확보하였고 이후에 효소 활성을 검사하였다. 인체에 악영향을 미칠 수 있 는 β-glucuronidase 효소는 활성이 나타나지 않았으며, 시중에서 판매되고 있는 Bacillus 프로바 이오틱스와 유사한 효소의 활성과, 추가적으로 펩타이드 가수분해효소인 Leucine arylamidase 의 활성이 나타난 NB-1 균주가 확인되었다. 이에, 추가적인 효능 연구 및 안정성 실험을 통해 프로바이오틱스 균주로서의 가능성을 모색해야 할 필요가 있다고 판단된다.

This research focused on the isolation and characterization of the new strains which can be used as microbial resources for auxin production in the agriculture industry. For the isolation of the new microorganism in the gut of Larimichthys polyactis, a marine agar medium was used and 3 colonies were isolated. Through the 16S-based ID service, isolated strains were identified as 1 strain of Niallia circulans and 3 strains of Proteus mirabilis. Verifying the agriculture industrial values of these isolated strains, the productivity of auxin and activity of various enzymes such as amylase, lipase, and protease were confirmed. As a result, isolated 3 strains showed auxin activity and only protease activities which means the possibility of applying them to the production of effective microorganisms in the agriculture industry.

Screening of Bacillus subtilis strains capable of producing extracellular hydrolases, including lipolytic enzyme was carried out. Within the scope of this study, total 61 strains were isolated from various domestic specific environmental samples such as seawater, soil, Jeju ranch, hot spring, salt farm, and fermented food and identified as Bacillus subtilis based on 16S rRNA sequences. Among the isolates, 4 strains had extracellular lipase activity. The growth profile of the strains revealed that 4 strains showed well growth at 50℃, 3 strains at 55℃, and pH 7.0. All strains could tolerate salinity up to 5%(w/v), 3 strains up to 10%(w/v), and only one strain showed growth at 15%(w/v). Additionally, amylolytic and proteolytic activities were detected in these strains. The highest lipolytic, amylolytic, and proteolytic activity was detected in Bacillus subtilis YS-YR 5A. These results demonstrate the potential application of extracellular hydrolase-producing Bacillus subtilis strains, especially strain YS-YR 5A, as fermentation starter to enhance the functionality and multiplication of functional natural products in the food/medicine/cosmetics/bio industries.

In probiotics production, lactic acid bacteria (LAB) are cultured on a large scale to achieve efficient processing through fermentation optimization and scale-up. In this Current study, the LAB strain Lactiplantibacillus plantarum WiKim0125 was isolated from kimchi and optimized the production. The optimization strategy for cultivating L. plantarum WiKim0125 consisted of investigating media components, selecting physiochemical conditions to enhance productivity, and scaling up for pilot-scale production. Each process condition was evaluated based on substrate consumption, lactic acid production, and viable cell yield. As a result, the final viable cell and lactic acid yield of L. plantarum WiKim0125 increased by 38.6% and 19.4%, respectively. This study provides an overview of fermentation optimization and scale-up processes for the industrial application of L. plantarum WiKim0125.

Dental caries is an infectious disease accompanied by the destruction of teeth. It is a multifactorial disease caused by the interaction between microbes in dental plaque and food, with cariogenic Streptococcus mutans being one of the primary causes. In this study, the antimicrobial and antibiofilm activities of Lactiplantibacillus plantarum THK-j112 isolated from Kimchi, a traditional fermented food, against S. mutans were investigated. Furthermore, this paper also showed the possibility of developing this strain. After analyzing the 16S rRNA sequence, strain THK-j112 was named L. plantarum THK-j112 as it shared 99.93% similarity with L. plantarum ATCC 14917T. Both L. plantarum THK-j112 and L. plantarum ATCC 14917T demonstrated MIC values of 5 mg/mL and MBC values of 10 mg/mL against S. mutans KACC 16833T. The study confirmed that L. plantarum THK-j112 exhibited significant concentration-dependent inhibitory effects on biofilm formation, acid production, and glucosyltransferases(GTFs) production. Regarding hemolytic activity, Bacillus cereus ATCC 14579T, used as a positive control, was confirmed to cause hemolysis, whereas L. plantarum THK-j112 did not cause this phenomenon. Additionally, except for gentamicin, the antibiotic resistance test revealed a lower value than the MIC cut-off value for each antibiotic, indicating no resistance. Based on the findings, L. plantarum THK-j112 is expected to be useful in the food industry, such as functional food, as a lactic acid bacterium with antimicrobial and antibiofilm effects.

Exposure to lipopolysaccharide (LPS) causes cognitive impairment (CI). In the preliminary study, Lactobacillus gasseri NK109 suppressed LPS-induced expression of proinflammatory cytokines in macrophages. Therefore, the effect of NK109 on LPS-increased CI was investigated in mice. Intraperitoneal injection of LPS caused CI-like behaviors and neuroinflammation. However, orally administered NK109 reduced LPS-increased CI-like behaviors and hippocampal IL-1β and TNF-α expression, whereas LPS-decreased BDNF expression increased. NK109 also reduced LPS-increased colonic myeloperoxidase, IL-1β, and TNF-α expression. The efficacy of NK109 was increased by the combination of soybean embryo ethanol extract (SE). These findings suggest that NK109 with SE can improve CI by alleviating inflammation-mediated BDNF expression, thereby being beneficial for dementia therapy.

To enhance the bioavailability and bioactivities of mixed herbal medicines (RW), they were fermented with lactic-acid bacteria isolated from kimchi into postbiotics (FRW). Then, from the results of the 16s rRNA sequencing analysis, lactic acid bacteria isolated from kimchi were identified to be of two species, namely Lactobacillus sakei and Leuconostoc mesenteroides. The FRW prepared from the RW were extracted using hot water (HW) and 70% EtOH (EtOH) for comparison of their macrophage-stimulating activities. Based on a comparison of the activities of the FRW extracts, nitric oxide (NO) production of HW was significantly higher than that in EtOH. An analysis of the chemical properties of the extracts showed that HW had higher contents of neutral sugar and uronic acid than EtOH as well as contained a large amount of glucose. In addition, crude polysaccharide (CP) was prepared to enhance the macrophage-stimulating activity. The FRW-CP not only secreted immunostimulatory mediators but also increased the expression of immunostimulatory genes (iNOS, TNF-α, MCP-1, and IL-6). The fractionated FRW-CP contained about 90% neutral sugars, and these sugars were mainly composed of glucose, galacturonic acid, and arabinose. Thus, FRW prepared by fermentation of RW with kimchi lactic acid bacteria were found to be immunostimulatory modulators.

In this study, probiotic properties such as acid resistance, bile tolerance, intestinal adhesion and fat accumulation inhibition of lactic acid bacteria isolated from kimchi were investigated in order to develop probiotics effective anti-adipogenic effects. Among the 120 types of lactic acid bacteria isolated from kimchi, 42 types of lactic acid bacteria being to acid resistance, 35 types of lactic acid bacteria being to bile resistance, and 24 types of lactic acid bacteria being to both acid resistance and bile resistance were selected. These 24 types were tested for the intestinal adhesion ability which is an essential factor of probiotics’ function and L. rhamnosus GG was used as a positive control in this test. 10 types of lactic acid bacteria having the intestinal adhesion ability were selected. The selected 10 types of lactic acid bacteria were treated in 3T3-L1 cells and intracellular fat was stained with Oil Red O reagent to measure fat accumulation. As a result, in the group treated with lactic acid bacteria OS-15, 48, 102 and 120, fat accumulation was significantly decreased compared with the non-treated group. And the strain OS-48 showed the greatest inhibitory effect on fat accumulation. The lactic acid bacterium OS-48 was identified as Lactiplantibacillus plantarum by analysis based on 16S rRNA sequencing and was named Lactiplantibacillus plantarum OS-48. Therefore, based on the results of this study, L. plantarum OS-48 is expected to be very useful as probiotics with anti-obesity effect in the food industry such as fermented food and functional food.

본 연구에서는 김치로부터 분리한 유산균 Weizmannia coagulans KK7(KCTC19023P) 균주를 이용하여 당근(Daucus carota var. sativa) 잎 추출물의 발효를 실시하고 얻은 생산물의 항염증 활성 및 성 분변화를 확인하였다. 발효에 의한 당근 잎 추출물의 성분 변화를 확인하기 위하여 고성능 액체크로마토그 래피를 이용하여 분석한 결과 발효 전과 비교하였을 때 발효 후에 플라보노이드의 일종인 루테올린 (luteolin)의 함량이 현저히 증가하는 것을 확인할 수 있었다. 당근 잎 추출물과 발효 당근 잎 추출물의 항 염 활성은 LPS에서 염증을 유발한 RAW 264.7 세포에서 NO (Nitric oxide) 생성 억제 효과로 확인하였 다. 발효 당근 잎 추출물을 처리하였을 때 당근 잎 추출물 처리군에 비해 NO 생성이 상대적으로 억제되는 것을 확인할 수 있었다. 발효 당근 잎 추출물 농도가 증가함에 따라 NO 생성 억제 효과도 유의적으로 증 가 하였다. Western blot을 통해 염증반응에서 NO 생합성과 관련된 효소 iNOS의 발현을 확인한 결과 발 효 당근 잎 추출물에서 단백질 발현이 농도 의존적으로 감소하는 것을 확인할 수 있었다. 이러한 실험 결과 로부터 미생물 발효를 통해 당근 잎의 항염 효과가 증대된다는 것을 확인할 수 있었으며 이는 식품으로 거 의 이용되지 않고 버려지는 당근 잎이 발효를 통해 새로운 식품 및 화장품 소재로 사용될 수 있음을 시사 한다.

본 연구에서는 미생물 첨가에 따라 거세한우 비육우 분의 이화 학적 특성, 미생물 성상, 가스발생량 및 퇴비 부숙도에 미치는 영 향을 규명하고자 수행하였다. 이상의 결과를 종합하면, 4주 후,미생물 첨가구에서 수분, 유기물, 총질소 함량 및 pH가 낮았으며, 나머지 이화학적 특성에서는 차이가 나타나지 않았다. 유산균과 효모균 수는 증가하였으며, 대장균 수는 감소하였다. 12주 후, 미 생물 첨가구에서 유산균과 고초균 수는 높았으나, 수분, 효모 및 대장균 수는 낮게 낮았다. 하지만 암모니아, 황화수소 발생량과 퇴비 부숙도는 미생물 첨가에 의한 효과가 나타나지 않았다. 따라 서, 거세한우 분에 미생물을 첨가하면 유익균은 증가하고, 병원성 미생물은 감소하여, 비육우의 생산성은 증진될 것으로 사료되지 만, 가스 발생량 및 퇴비 부숙도에 대한 추가적인 연구는 지속적 으로 수행되어야 할 것으로 사료된다.

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.

After liquid culture of Phellinus baumii (P. baumii) mycelium (LPBM) was prepared, LPBM was fractionated into A∼E fraction (A; hot-water extract of liquid culture including mycelia, B; crude polysaccharide of A, C; hot-water extract of mycelia, D; crude polysaccharide of C, and E; crude polysaccharide of culture broth) to evaluate for possibility as functional materials with immunostimulatory activity. In macrophage stimulatory activity, E fraction as postbiotics significantly increased secretion of NO and IL-12 from RAW 264.7 cells. Next, when the splenocytes of C3H/HeN mice were primary cultured, E fraction showed significantly mitogenic activity with enhancing mitogen-related cytokines (IFN-γ and TNF-α) production from splenocyte. E fraction also potently stimulated GM-CSF production from Peyer’s patch cells as well as Peyer’s patch-mediated bone marrow cell proliferation. In addition, the immunostimularoy E fraction contained neutral sugar (73.8%), uronic acid (10.6%), protein (7.8%), and polyphenol (7.5%), and mainly consisted of glucose (39.1%), galactose (21.7%), mannose (11.1%), galacturonic acid (9.9%), and arabinose (8.9%) as component sugars. In conclusion, it was demonstrated that postbiotics including exopolysaccharide fractionated from liquid culture of the P. baumii mycelium could enhanced immunostimulatory activity.

This study aimed to isolate and identify the new strains which can be utilized in fermentation process for the production of functional materials. Isolation of the new microorganism in wood vinegar, natural liquid material generated from the carbonization of various trees was studied. Marine agar medium was used for the isolation of halo-tolerant bacteria. Using a sterilized toothpick, transfer same shape colonies to agar plates and continuous cultivation of colonies at 37℃ for several days, 2 colonies were isolated. Through the 16S-based ID service, isolated strains were identified as Cytobacillus species. Verifying the industrial values of the two isolated strains, the productivities of various enzymes such as amylase, lipase, and protease were confirmed. As a result, isolated 2 strains showed amylase and protease activities which means the possibility of applying to biological processes in the food and cosmetic industries.

Liquid-state fermentation (LSF) and solid-state fermentation (SSF) were carried out to produce enzymes and β-glucan. The modified synthetic medium composition in liquid-state fermentation (LSF) was determined to be 5% (w/v) whole oat flour, 2% (w/v) tryptone, and 2% (w/v) rice bran. The different humidity conditions of oat by solid-state fermentation were evaluated in terms of maximum production of fungal biomass, amylase, protease, and β-glucan, which were 0.23 mg/g, 7,157.38 U/g, 413.67 U/g, and 8.26% (w/w), respectively, at 60% of humidity condition. Moreover, α-glucosidase inhibition activities of fermented oat with 60% humidity at 48 h were shown to be high 49.08% (w/w). Therefore, liquid- and solid-state fermentation processes were found to enhance the overall fermentation yields of oat to produce enzymes and β-glucan.

The aim of this study was to evaluate the probiotic properties of lactic acid bacteria (LAB) isolated from kimchi and antioxidant and anti-inflammatory activities of heat-killed LAB strains. Regarding probiotic properties, Lactiplantibacillus plantarum SMF398 and L. plantarum SMF470 isolated from kimchi can survive under artificial gastric condition and adhere strongly to HT-29 cells. The SMF398 and SMF470 strains showed strong antimicrobial activity and co-aggregation against pathogenic bacteria. The heat-killed cells of LAB (1 mg/mL) were prepared by heating at 121℃ for 15 min. MTT assay revealed that the heat-killed cells (1 mg/mL) of SMF398 and SMF470 were not toxic to HT-29 cells. The heat-killed SMF398 and SMF470 showed significantly higher DPPH and ABTS radical scavenging activities as well as β- carotene bleaching inhibitory activity than the heat-killed L. plantarum ATCC14917, a control probiotic strain (p<0.05). In lipopolysaccharide-induced RAW264.7 cells, the heat-killed SMF398 and SMF470 significantly reduced the nitric oxide production by 30.92% and 22.81%, respectively (p<0.05). Furthermore, the heat-killed SMF398 and SMF470 significantly decreased the gene expression levels of tumor necrosis factor-α, interleukin-6, inducible nitric oxide synthase, cyclooxygenase-2 up to 58.02%, 67.70%, 53.44%, 45.90%, respectively (p<0.05). These results suggest that the heat-killed L. plantarum SMF398 and L. plantarum SMF470 might be useful as antioxidant and anti-inflammatory agents.

This study aimed to investigate the effects of antioxidant and anti-inflammatory activities of heat-killed lactic acid bacteria (LAB) produced under different temperature conditions. Regarding probiotic properties, Limosilactobacillus fermentum SMF743 and Lactiplantibacillus plantarum SMF796 isolated from kimchi showed strong acid and bile salt resistance, adhesion activity onto HT-29 cells, and antimicrobial activity against pathogenic bacteria. Based on the results of thermal death time and temperature, heat-killed LAB cells (1 mg/mL) were prepared by heating at 70oC (180 min), 80oC (120 min), 100oC (30 min), and 121oC (15 min). The heat-killed SMF743 and SMF796 showed significantly higher DPPH and ABTS radical scavenging activities than live cells (p<0.05). The heat-killed SMF743 and SMF796 at 70oC or 121oC revealed stronger DPPH and ABTS radical scavenging activities and inhibition of nitric oxide production than those at 80oC or 100oC. Furthermore, heat-killed SMF743 and SMF796 at 121oC significantly reduced the gene expression levels of tumor necrosis factor-, inducible nitric oxide synthase, and cyclooxygenase- 2 up to 53.33%, 58.67%, and 83.67%, respectively, in lipopolysaccharide (LPS)-induced RAW264.7 cells (p<0.05). These results suggest that heat-killed L. fermentum SMF743 and L. plantarum SMF796 can be used as natural antioxidants and anti-inflammatory agents.

본 연구의 목적은 효소 가수분해 및 유산균 혼합발효기술을 이용하여 GABA 함량이 강화된 발효 다시마 추출물을 생산함에 있다. TLC 분석방법을 통해 GABA 생성능력이 우수한 균주 L. plantarum KCTC 21004, L. acidophilus KCTC 3164, L. sakei subsp. sakei KCTC 3598를 선정하였 다. 선정된 균주를 이용한 유산균 발효의 특성은 배양 시간이 증가 할수록 적정 산도는 증가하였고 pH 는 감소하는 경향을 나타내었다. GABA 생성능력이 우수한 3종의 균주 중 L. plantarum KCTC 21004 이 GABA 함량이 136.4 mg/100g으로 우수하였다. 이러한 연구 결과는 유산균을 이용한 GABA 생산과 정에 있어서 기초적인 자료로 제공될 것으로 판단된다.