• 한국식품과학회지
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• 제36권6호
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• pp.959-967
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• 2004

인간의 대장내 점막에 대하여 우수한 점착특성을 갖는 probiotic 유산균주를 선별할 목적으로, colonic mucin-binding assay를 고안하고 최적의 분석 조건을 검토한 결과, microtiter plate의 well에 대한 colonic mucin의 부착은 pH 4.8, biotinylated SLP의 농도는 $5.0\;{\mu}g/mL$, 시판 HRP-conjugated streptoavidin은 24,000배 희석용액, TMB의 발색시간은 10분의 조건에서 측정시 최적의 결과를 나타냈다. 동 조건에서 본 assay system을 이용할 경우, 장내 점착능 측정 및 우수 유산균주의 선별에 있어 신속하고 재현성 있는 결과를 제공할 수 있으며, 인간의 대장에 대한 유산균의 점착특성을 정량적으로 분석할 수 있음을 보여주었다. 공시균주 32종 및 유아 분변 유래의 분리균주 18종을 포함한 총 50종의 유산균주에 대하여, colonic mucin-binding assay를 이용하여 대장 mucin에 대한 결합능을 비교한 결과, L. species FSB-1이 가장 높은 결합능을 보여주었다. 따라서 L. species FSB-1을 대상으로 형태학적 특성, 생리 및 생화학적 특성과 16S rDNA에 대한 부분 염기서열 분석을 포함한 동정실험을 수행한 결과, 장내 점착능 우수균주로 선별된 L. species FSB-1은 Lactobacillus brevis로 최종 동정되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제21권10호
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• pp.1495-1500
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• 2008

In an attempt to develop a probiotic formulation for poultry feed, a number of lactic acid bacteria (LAB) were isolated from chicken intestinal specimens and a series of in vitro experiments were performed to evaluate their efficacy as a potential probiotic candidate. A total of 650 LAB strains were isolated and screened for their antagonistic potential against each other. Among all the isolates only three isolates (TMU121, 094 and 457) demonstrated a wide spectrum of inhibition and were thus selected for detailed investigations. All three selected isolates were able to inhibit the growth of E. coli and Salmonella species, although to variable extent. The nature of the inhibitory substance produced by the isolates TMU121 and 094 appeared to be associated with bacteriocin, as their activity was completely lost after treatment with proteolytic enzymes, while pH neutralization and catalase enzyme had no effect on the residual activity. In contrast, isolate TMU457 was able to resist the effect of proteolytic enzymes while pH neutralization completely destroyed its activity. Attempts were made to study the acid, bile tolerance and cell surface hydrophobicity of these isolates. TMU121 showed high bile salt tolerance (0.3%) and high cell surface hydrophobicity compared to the other two strains studied, while TMU094 appeared the most pH resistant strain. Based on these results, the three selected LAB isolates were considered as potential ingredients for a chicken probiotic feed formulation and were identified to species level based on their carbohydrate fermentation pattern by using API 50CH test kits. The three strains were identified as Lactobacillus fermentum TMU121, Lactobacillus rhamnosus TMU094, and Pediococcus pentosaceous TMU457.

• 한국축산식품학회지
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• 제35권1호
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• pp.91-100
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• 2015

The present study was conducted to screen candidate probiotic strains for anti-inflammatory activity. Initially, a nitric oxide (NO) assay was used to test selected candidate probiotic strains for anti-inflammatory activity in cultures of the murine macrophage cell line, RAW 264.7. Then, the in vitro probiotic properties of the strains, including bile tolerance, acid resistance, and growth in skim milk media, were investigated. We also performed an in vitro hydrophobicity test and an intestinal adhesion assay using Caenorhabditis elegans as a surrogate in vivo model. From our screening, we obtained 4 probiotic candidate lactic acid bacteria (LAB) strains based on their anti-inflammatory activity in lipopolysaccharide (LPS)-stimulated RAW 264.7 cell cultures and the results of the in vitro and in vivo probiotic property assessments. Molecular characterization using 16S rDNA sequencing analysis identified the 4 LAB strains as Lactobacillus plantarum. The selected L. plantarum strains (CAU1054, CAU1055, CAU1064, and CAU1106) were found to possess desirable in vitro and in vivo probiotic properties, and these strains are good candidates for further investigations in animal models and human clinical studies to elucidate the mechanisms underlying their anti-inflammatory activities.

• Journal of Microbiology and Biotechnology
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• 제12권4호
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• pp.610-616
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• 2002

The growth-inhibiting effects of Pinus densiflpora leaf-derived materials on nine human intestinal bacteria were investigated using the impregnated paper disk method, and their activities were compared with those of 13 commercially available terpenes. The biologically active constituent of the extract of P densiflora leaf was characterized as the monoterpene (1R)-(+)-$\alpha$-pinene by various spectroscopic analyses. Responses varied according to bacterial strain, chemicals, and dose. At 10 mg/disk, limonene and (1R)-(+)-$\alpha$-pinene strongly inhibited the growth of Clostridium perfringens, Staphylococcus aureus, and Escherichia coli, without adverse effects on the growth of five lactic acid-bacteria (Bifidobacterium adolescentis, B. bifidum, B. longum, Lactobacillus acidophilus, and L. casei). Little or no inhibition against seven bacteria was observed with anethole, borneol, camphor, caryophyllene, 1,8-cineole, estragole, linalool, and $\alpha$-terpineol. Structure-activity relationship revealed that (1R)-(+)-$\alpha$-pinene had more growth-inhibiting activity against C. perfringens than (1R)-(+)-$\beta$-, (1S-(-)-$\alpha$-, and (1S-(-)-$\beta$-pinenes. Furthermore, the growth-inhibition against L. casei was much more pronounced in (1R)-(+)-$\beta$- and (In-(-)-$\beta$-pinenes than (1R)-(+)-$\alpha$- and (1S)-(-)-$\alpha$-pinenes. These results indicate that the (+)-$\alpha$ form seems to be required against C. perfringens and $\beta$ form against L. casei for growth-inhibiting activity. Morphologically, most strains of C. perfringens were damaged and disappeared at 5 and 2 mg/disk of (1R)-(+)-$\alpha$-pinene. Morphological study revealed that (1R)-(+)-$\alpha$-pinene had more growth-inhibiting activity against C. perfringens than (1R)-(+)-$\beta$-, (1S)-(-)-$\alpha$-, and (1S)-(-)-$\beta$-pinenes. As naturally occurring growth-inhibiting agents, the Pinus leaf-derived materials described above could be useful preventive agents against diseases caused by harmful intestinal bacteria such as clostridia.

• Food Science and Biotechnology
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• 제14권4호
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• pp.523-528
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• 2005

The commercial interests in functional foods containing probiotics are gaining significance in view of increasing studies on their role in digestive tract. Probiotic dairy foods containing health-promoting bacteria are important segment of functional food market. Various health benefits have been attributed to specific strains of lactic acid bacteria or food containing these probiotic cultures. Probiotic-containing foods are considered to improve general gut health and natural defenses of body, and lower blood cholesterol level. Specific probiotic microbes can alleviate or prevent diverse intestinal diarrhea-inducing disorders, cause prophylaxis of intestinal and urogenital infections, inhibit mutagenicity of intestinal contents, and reduce incidence of intestinal tumors. Recent increasing evidences on health effects of probiotics have triggered consumer interest in this category of functional foods. Rational approach needs to be applied in selection of strains for probiotic preparation to achieve required functionality. Present article focuses on some prominent probiotic candidates and criteria for their inclusion in functional food sector. Various health claims of probiotics on gastrointestinal disorders, anticarcinogenic effects, and anti-cholestrimic effects, and possible mechanistic explanations for their functionality are highlighted.

• 한국식품영양과학회지
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• 제46권7호
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• pp.857-867
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• 2017

본 연구는 저분자 해양성 콜라겐과 GABA 생성 L. brevis CFM20의 섭취 시 다양한 인체 장내 조건에 의한 유산균의 사멸과 콜라겐의 분해를 최소화하고자 진행되었다. 유산균과 저분자 해양성 콜라겐의 안정성을 향상하기 위하여 alginate와 chitosan을 이용하여 이중코팅캡슐을 제조하였다. L. brevis CFM20의 온도별 생육특성을 분석하여 캡슐화에 유리한 배양온도를 조사한 결과 $37^{\circ}C$에서 배양한 균의 대수기가 가장 빠르고 GABA양 또한 $400{\mu}g/mL$로 가장 높은 수치를 나타내어 생육속도에서 가장 유리한 것으로 나타났다. Calcium-alginate bead 법으로 캡슐을 제조한 결과 1~2 mm 사이의 일정한 구형의 캡슐을 제조하였으며, chitosan을 이용하여 이중코팅캡슐을 제조한 결과 캡슐의 크기가 감소하였고 외형적으로도 더욱 단단해짐을 확인하였다. 제조된 이중코팅캡슐을 CLSM과 SEM을 이용하여 관찰한 결과, chitosan 코팅을 한 이중코팅캡슐의 표면이 alginate와의 가교결합에 의해 더욱 치밀해짐을 확인하였다. 체내 위와 장내 조건에서 시간에 따른 이중코팅캡슐의 L. brevis CFM 20 생균수를 분석한 결과 캡슐화한 경우 균의 감소가 더디어 더 안정하다고 판단하였다. 이중코팅캡슐을 체내 위와 장내 조건에서 시간에 따른 GABA와 hydroxyproline의 용출량을 분석한 결과 두 경우 모두 위 조건에서는 낮은 수치를 나타내었고 장내 조건에서는 시간이 지남에 따라 급격하게 증가한 것을 확인하였다. 동물실험 결과 콜라겐 섭취군이 비섭취군에 비해 체중증가량, 혈청 지질 농도 등이 적은 것을 확인하였고, 혈액 내 hydroxyproline 함량, 진피의 피부조직 밀도가 더 높은 것으로 관찰되었다. 본 연구에서 개발된 이중코팅캡슐을 섭취하는 경우 위에서는 안정한 상태였다가 장에 도달한 후 붕해됨으로 인하여, 유산균, GABA, 콜라겐 및 키토산의 작용으로 인체에 유익한 효과를 기대할 수 있을 것으로 생각된다.

• Journal of Dairy Science and Biotechnology
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• 제38권2호
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• pp.70-88
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• 2020

대부분의 스타터 배양균은 유산균(LAB)군(group)에 속하며 미국 식품의약국(FDA)과 유럽식품안전청(EFSA)에 의해 안전하다고 인정받고 있다. 그러나, LAB는 항생제 내성(antibiotic resistance) 유전자를 위한 내적 또는 외적 저장소로 작용할 수 있다. 하지만 저항 유전자 전달이 수직이기 때문에 이 사실은 그 자체가 안전상의 문제가 아닐 수도 있다. 그럼에도 불구하고 외부 유전적 요소는 심각한 안전 문제를 나타내는 인체 장내 미생물뿐만 아니라 병원균의 저항성 수평 전달에 유리한 변화를 유도할 수 있다. 항생제 내성을 가진 유산균의 일부 속(genus)에는 발효육 및 유제품에서 분리된 Enterococcus, Lactobacillus, Lactococcus, Leuconostoc, Pediococcus, Streptococcus 등이 포함된다. 현재 WHO는 식품산업에서 사용되는 LAB는 저항이 없어야 한다고 권고하고 있다. 다행히도 한국의 식품의약품안전처가 인정한 프로바이오틱스 균주 중 Enterococcus에 해당되는 균주는 E. faecium과 E. faecalis인데, 2019년 4월 1일부터 E. faecium과 E. faecalis의 독성인자와 항생제 내성전달인자 모두 없다고 인정된 경우에면 사용이 허가된다. 따라서 LAB의 산업적 중요성을 고려할 때 앞으로 인체와 동물장관 유래의 LAB은 체계적인 탐색 및 동정, 병원성 및 항생제 내성 유전자의 확인, 식품 및 가축사료 첨가제로의 활용, 안전성 평가연구는 계속 필요하다고 사료된다.

• Journal of Microbiology and Biotechnology
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• 제13권4호
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• pp.522-528
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• 2003

The growth responses of Phellodendron amurense root-derived materials against seven intestinal bacteria were examined, using an impregnated paper disk agar diffusion method and spectrometric method under $O_2$-free condition. The biologically active constituent of the P. amurense root extract was characterized as berberine chloride ($C_{20}H_{18}NO_{41}Cl$) using various spectroscopic analyses. The growth responses varied depending on the bacterial strain, chemicals, and dose tested. At 1 mg/disk, berberine chloride strongly inhibited the growth of Clostridium perfringens, and moderately inhibited the growth of Escherichia coli and Streptococcus mutans without any adverse effects on the growth of three lactic acid-bacteria (Bifidobacterium bifidum, B. longum, and Lactobacillus acidophilus). The structure-activity relationship revealed that berberine chloride exhibited more growth-inhibiting activity against C. perfringens, E. coli, and S. mutans than berberine iodide and berberine sulfate. These results, therefore, indicate that the growth-inhibiting activity of the three berberines was much more pronounced as chloridated analogue than iodided and sulphated analogues. As for the morphological effect caused by 1 mg/disk of berberine chloride, most strains of C. perfringens were damaged and killed, indicating that berberine chloride showed a strong inhibition against C. perfringens. As naturally occurring growth-inhibiting agents, the P. amurense root-derived materials described could be useful as a preventive agent against diseases caused by harmful intestinal bacteria such as clostridia.

• 대한의생명과학회지
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• 제16권4호
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• pp.213-220
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• 2010

Although an adequete intake of calcium (Ca) is recommended for the treatment and prevention of osteoporosis, the intake of Ca should be restricted because of its low rate of intestinal absorption. The purpose of this experiment was to identity the effect of the combined administration of Aquamin F (AQF) (a calcium agent) and lactic acid bacteria (LAB) on osteoporosis. Thirty ovariectomized (OVX) rats and six control rats were assigned to the following six groups, with six animals per group: sham Ca-deficient diet (Ca-D), OVX, LAB, AQF, and LAB-AQF. During the experiment, the body weight was measured; and after the experiment was completed, the serum biochemical analysis, the alkaline phosphatase, calcium, and inorganic phosphorus leves were measured. The tissue of the femur was stained and then scanned via CT. The body weight of the OVX group increased more significantly than that of the control group. The results of the bone mineral content (BMC), Bone mineral density (BMD), serum biochemical analysis and histological test on the femur epiphysis showed no difference between the OVX group and the LAB group, whereas the results of the AQF group were more significant than those of the OVX group. In particular, the LAB+AQF group showed more significant increases in the aforementioned results than the AQF group. This experiment showed that the combined administration of AQF and LAB in ovariectomized rats more significantly increased bone density than did a single administration of either AQF or LAB.

• Journal of Animal Science and Technology
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• 제52권2호
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• pp.111-116
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• 2010

본 연구는 말의 다양한 사양체계가 분 젖산 생산 박테리아 및 pH 변화에 미치는 영향을 구명하기 위해 수행되었다. 시험 1에서는 농후사료 체중 2% 또는 건초(오챠드그라스) 2%를 30일 동안 급여한 후 분 중 Lactobacillus와 Streptococcus 균 수를 측정했다. 시험 2에서는 사양형태 또는 사료급여 수준에 따른 분 중 pH 변화를 측정했다. 시험 3에서는 알팔파 건초 급여 수준에 의한 분 pH 변화를 측정했다. 고－농후사료의 급여는 분 중 젖산 생산 박테리아인 lactobacillus와 streptococcus의 균 수를 크게 증가시켰다(P<0.05). 농후사료의 급여량을 증가시킴에 따라 분 pH는 유의적으로 감소했다(P<0.01). 알팔파 건초 단독 급여에서는 분 pH가 저하되지 않았으나, 알팔파 건초+농후사료 사양형태에서는 분 pH가 감소되었다(P<0.01). 본 연구 결과, 말에게 농후사료의 다량급여는 장내 젖산생산 박테리아의 활성을 증가시키며, 농후사료 급여량이 증가함에 따라 분 pH를 저하시키고 있음을 알 수 있다. 따라서 말에게 적절한 조사료/농후사료 비율을 유지하고 급여량을 조절하는 것이 필요하다. 또한 분 pH 측정으로 인해 대장 내 상태를 가늠할 수 있고, 분 pH 값을 이용해 사양관리의 지표로 활용할 수 있을 것으로 사료된다.