• Applied Biological Chemistry
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• 제35권3호
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• pp.179-185
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• 1992

우리나라 남부지방(南部地方)의 5개(個) 산지(山地) 목초지(牧草地)에 분포한 알팔파근류균(根瘤菌) R. meliloti의 밀도(密度)와 실소고정력(室素固定力), 그리고 항생제(抗生劑) 반응(反應) 특성(特性)을 조사하였다. 그 결과(結果), 초지토양(草地土壤)에 분포한 R. meliloti 밀도(密度)는 $1.7{\times}10^2\;cells/g$.토양(土壤)(여주시(麗州市) 충효동(忠孝洞)) 내지 $1.0{\times}10^5\;cells/g$.토양(土壤)(경주군(慶州郡) 감포면(甘浦面)) 수준(水準) 이었으며, 토양 pH와는 정(正)의 상관(相關)을 보였다. 그리고, 초지토양(草地土壤)의 알팔파 접종효과(接種效果)로 본 토착(土着) R. meliloti의 질소고정력(窒素固定力)은 전반적으로 표준균주(標準菌株)인 TAL 접종제(接種劑) 보다 저조(低調)하였으나, 달성군(達城郡) 옥포면(玉浦面) 초지(草地)에서 분리(分離)된 YCKa 539, YCKa 542 2균주(菌株)는 비슷하거나 우수(優秀)한 질소고정력(窒素固定力)을 보였다. 초지토양별로 보면 경주군(慶州郡) 견곡면(見谷面)의 것이 가장 높았고, 다음으로 달성군(達城郡) 옥포(玉浦)>경주시(慶州市) 충효(忠孝)>달성군(達城郡) 화국(花國)>경주군(慶州郡) 감포(甘浦) 균주(菌株) 순(順)이었다. 분리균주(分離菌株)들은 항생제(抗生劑) Ery($100\;{\mu}g/ml$), Nal($160\;{\mu}g/ml$), Str($10\;{\mu}g/ml$)에 대해 각각(各各) 67%, 77%, 80%정도의 내성경향(耐性傾向)을 보이면서 전체적으로는 14개(個)의 항생제(抗生劑) 내성류형(耐性類型)으로 분류(分類)되었는데, 이들 류형(類型)과 균주(菌株)의 질소고정력간(窒素固定力間)에는 뚜렷한 관계경상(關係傾商)을 보이지 않았다. 그리고, 분리균주(分離菌株) 중(中)에서 질소고정력(窒素固定力)이 가장 우수(優秀)하였던 YCKa 542균주(菌株)는 Ery Nal, Neo, Str 내성인 X번형(番型)에 속하였다.

• 한국축산식품학회:학술대회논문집
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• 한국축산식품학회 2004년도 정기총회 및 제33차 춘계 학술대회
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• pp.89-119
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• 2004

This study was conducted to isolate lactobacilli having probiotic characteristics to be used as health adjuncts with fermented milk products. Acid tolerant strains were selected in Lactobacilli MRS broth adjusted to pH 4.0 from 80 healthy persons (infants, children and adults). And bile tolerant strains were examined in Lactobacilli MRS broth in which 1.0% bile salt was added. By estimation above characteristics, the strains No. 27, which was isolated from adult feces, was selected and identified as Lactobacillus salivarius subsp. salivarius based on carbohydrate fermentation and 16S rDNA sequencing. It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at $37^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by $0.9{\sim}1.0%$ at $37^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced $23{\sim}38%$ of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 hours at $37^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with $15{\sim}25$ mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. Purified ${\alpha}$-galactosidase was obtained by DEAE-Sephadex A-50 ion exchange chromatography, Mono-Q ion exchange chromatography and HPLC column chromatography from L. salivarius subsp. salivarius 27. The specific activity of the purified enzyme was 8,994 units/mg protein, representing an 17.09 folds purification of the original cell crude extract. The molecular weight of enzyme was identified about 53,000 dalton by 12% SDS-PAGE. Optimal temperature and pH for activity of this enzyme were $40^{\circ}C$ and 7.0 respectively. The enzyme was found to be stable between 25 and $50^{\circ}C$. ${\alpha}$-galactosidase activity was lost rapidly below pH 5.0 and above pH 9.0. This enzyme was liberated galactose from melibiose, raffinose, and stachyose, and also the hydrolysis rate of substrate was compound by HPLC. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.