• 한국축산식품학회지
• /
• 제26권1호
• /
• pp.127-135
• /
• 2006

우유와 두유를 2:1로 혼합하고 홍삼 추출물을 첨가하여 발효유를 제조하였고 2종류의 스타터를 사용하여 발효 특성을 시험하였다. 2종류의 스타터를 사용한 모든 처리구에서 배양 15시간 후 pH는 $3.90{\sim}3.94$ 사이로 나타나 산 생성 촉진 효과가 있었고, 산 생성은 홍삼 추출물 첨가량이 증가함에 따라 조금씩 증가되는 것으로 나타났다. 홍삼 추출물 첨가량이 증가할수록 생균수는 증가하여 Lactobacillus acidophilus KCTC 3150를 스타터로 사용한 균에서 1.0% 홍삼 추출물 첨가량은 $6.26{\times}10^8\;cfu/mL$로서 최대 균수를 나타내었으며, 배양 12시간 이후에 대조구에서는 감소하는 반면 홍삼 추출물 첨가구에서는 젖산균수는 계속해서 최대균수를 유지하고 있었다. 유기산 생성은 lactic acid인 경우 Lactobacillus acidophilus KCTC3150에서 1.0% 홍삼 추출물 첨가구에서 332.22 mM로 가장 높았으며 홍삼 추출물 첨가량이 증가됨에 따라 lactic acid의 생성량도 증가되었다. 당 분해율은 두 균주 모두 홍삼 추출물 첨가량이 증가함에 따라 stachylose의 분해율이 증가하였고, Lactobacillus acidophilus KCTC3150 균주의 경우 stachylose의 분해율이 최대 19%인 반면, Lactobacillus salivarius subsp. salivarius CNU27의 경우 stachylose의 최대 분해율은 54%로서 분해율이 월등히 높았다. 점도는 홍삼추출물 첨가량이 1.0% 첨가구에서 Lactobacillus acidophilus KCTC3150, Lactobacillus salivarius subsp. salivarius CNU27 각각 780 cP로 가장 높게 나타났다. 관능검사 결과는 0.2% 홍삼 추출물을 첨가하고 Lactobacillus salivarius subsp. salivarius CNU27를 스타터로 사용한 발효유가 $4.14{\pm}0.64$로 기호도가 가장 좋게 나타났다.

• Asian-Australasian Journal of Animal Sciences
• /
• 제15권12호
• /
• pp.1798-1807
• /
• 2002

This study was conducted to investigate lactobacillus salivarius subsp. salivarius having probiotic properties to be used as the health adjuncts with fermented milk products. Acid- and bile-tolerant lactobacillus salivarius subsp. salivarius was isolated with lactobacilli MRS broth from faeces of 80 healthy persons (infants, children and adults). 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 0.9-1.0% 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 23-38% of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 h at 37$^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with 15-25 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. 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.

• 한국식품영양과학회지
• /
• 제38권12호
• /
• pp.1811-1814
• /
• 2009

귀리추출물에 L. delbrueckii subsp. bulgaricus 또는 S. salivarius subsp. thermophilus를 단일균주로 발효한 것에 비하여 L. delbrueckii subsp. bulgaricus와 S. salivarius subsp. thermophilus를 혼합하여 발효하는 경우에는 미생물균수와 산 생성이 증가하였다. pH의 변화에서도 S. salivarius subsp. thermophilus의 경우가 L. delbrueckii subsp. bulgaricus보다 더 낮은 pH를 나타내었으며, L. delbrueckii subsp. bulgaricus와 S. salivarius subsp. thermophilus를 혼합하여 발효하는 경우에는 pH가 더 빠르게 감소하였다. 이러한 결과는 우유를 원료로 한 요구르트의 경우처럼 귀리추출물의 젖산발효에서도 L. delbrueckii subsp. bulgaricus와 S. salivarius subsp. thermophilus의 두 균주 간에 생육촉진 현상이 있음을 의미하는 결과로 해석될 수 있는데 그 정도는 매우 적었다. 본 실험에서 S. salivarius subsp. thermophilus의 미생물균수와 산 생성이 더 증가한 반면 pH는 더 낮은 이유는 L. delbrueckii subsp. bulgaricus에 비하여 귀리추출물에 더 잘 적응하였기 때문이라고 생각된다.

• Asian-Australasian Journal of Animal Sciences
• /
• 제14권8호
• /
• pp.1170-1178
• /
• 2001

This study was conducted to isolate lactobacilli having 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 human faeces. Bile tolerant strains were examined in Lactobacilli MRS broth in which 1.0% bile salt was added. Microhemagglutination tests using swine erythrocytes were performed to select lactobacilli having adherence properties to survive in the intestinal tract. By examination of these characteristics the strain Nam 27, which was isolated from adult faeces, was selected and identified as Lactobacillus salivarius subsp. salivarius based on carbohydrate fermentation and 16S rDNA sequencing.

• 한국축산식품학회지
• /
• 제25권4호
• /
• pp.483-493
• /
• 2005

우유와 두유를 혼합하여 요구르트를 제조하고자 스타터에 따른 발효물의 성상을 시험하였다. 4종류의 서로 다른 스타터를 접종한 모든 처리구에서 배양 15시간 후 pH는 lactic culture 2(B. longum, L. acidophilus, S. thermophilus)를 제외한 나머지 처리구에서 $4.16\~3.75$ 사이로 나타나 발효 촉진효과가 있었고, 산 생성은 우유와 두유의 홉합비가 2:1이고 lactic culture 1 [Lactobacillus delbrueckii ssp. bulgaricus(LB12), Streptococcus salivarius ssp. thermophilus(ST36)]을 스타터로 사용한 요구르트에서 가장 높게 나타났다. 생균수는 1:3 비율에서 15시간째 Lactobacillus salivarius subsp. salivarius CNU27을 스타터로 사용한 균에서 $2.17\times10^9 cfu/mL$로서 최대 균수를 나타내었다. 두유 혼합요구르트에서 생산된 유기산의 함량을 측정한 바, 유기산 생산 중 lactic acid인 경우 lactic culture 1 [Lacto-bacillus delbrueckii ssp. bulgaricus(LB12), Streptococcus salivarius ssp. thermophihus(ST36)]을 스타터로 사용했을 때 1:1에서 412.52 mM로 가장 높았다. Acetic acid인 경우는 혼합비 3:1에서 lactic culture 2(B. longum, L. acidophilus, S. thermophilus)가 394.01mM로 가장 높게 생산되었다. 당 분해율은 혼합비 1:3에서 Lactobacillus salivarius subsp. salivarius CNU27를 스타터로 사용한 요구르트에서 대조구와 비교해 stachyose가 $53.92\%$로 가장 많이 분해되었다. 점도는 두유 혼합비가 가장 높은 1:3 처리구에서 $1,300\~1,660 cP$ 로 가장 높게 나타났다. 관능검사 결과는 2:1 혼합비에서 Lactobacillus acidophilus KCTC3150를 스타터로 사용한 요구르트가 $4.17\pm0.69,$ 3: 1 혼합비에서 Lactobacillus salivarius subsp. salivarius CNU27를 스타터로 사용한 요구르트가 $4.00\pm0..58$으로 기호도가 가장 좋은 것으로 나타났다.

• 대한소아치과학회지
• /
• 제32권2호
• /
• pp.344-356
• /
• 2005

우리 인체에는 의학적으로 유용한 기능을 가진 세균들이 정상적으로 존재하며 구강내에도 독특한 기능을 나타내는 세균들이 상재한다. 본 연구에서는 치아우식증이 없는 소아의 타액에서 분리한 유산균 2주가 Streptococcus mutans에 의한 인공치태 형성과 혐기성 세균에 의한 휘발성 유황화합물 생성을 억제하는 것을 확인하고, API 50 CHL medium kit를 이용한 생화학적 검사와 16S rDNA sequencing으로 동정하여 다음과 같은 결과를 얻었다. 1. 분리균주는 2주 모두 그람양성 간균으로 과산화수소를 생성하였다. 2. 인공치태의 무게는 Streptococcus mutans의 단독 배양시 $124.4{\pm}30.4mg$이었으나, 분리균주와 병합 배양시에는 각각 $5.2{\pm}2.0mg,\;10.6{\pm}6.6mg$으로 현저하게 감소하였다(p<0.05). 3. Streptococcus mutans의 생균수는 단독 배양시 ml당 $3.4{\times}10^9$이었으나, 분리균주와 병합 배양시에는 각각 ml당 $4.6{\times}10^8$과 $2.4{\times}10^8$으로 감소하였다. 4. Fusobacterium nucleatum을 30분간 진탕한 후 측정한 상청액의 흡광도는 1.286이었으나, Fusobacterium nucleatum과 분리균주를 병합으로 30분간 진탕한 후 측정한 상청액의 흡광도는 각각 0.628과 0.497로 감소하였으며, 상호 결합 정도는 29.4%와 57.8%이었다. 5. Fusobacterium nucleatum의 단독 배양시 cysteine과 $FeSO_4$를 첨가한 배지를 가한 후 측정한 침전물의 배지 흡광도는 1.794이었으나, 분리균주와 병합 배양시 측정한 침전물의 배지 흡광도는 각각 1.144와 0.915로 감소하였으며, Porphyromonas gingivalis 단독 배양시 침전물의 배지 흡광도는 1.932이었으나 분리균주와 병합 배양시에는 침전물의 배지 흡광도가 각각 1.170과 1.266으로 감소하였다. 6. 분리균주를 API 50 CHL medium kit로 탄수화물 발효검사를 시행한 결과, 분리균주 1주는 Lactobacillus salivarius로, 다른 분리균주는 Lactobacillus delbrueckii subsp. lactis로 동정되었다. 7 분리균주를 16S rDNA partial sequencing으로 동정한 결과, 2주 모두 Lactobacillus salivarius subsp. salicinius와 유전자 유사치가 99.60%, 99.73%를 보여 Lactobacillus salivarius subsp. salicinius로 동정되었다. 이상의 결과를 종합하면 치아우식증이 없는 소아의 타액에서 분리된 유산균 중 과산화수소를 분비하여 인공치태 형성과 휘발성 유황화합물 생성을 억제하는 분리균주는 Lactobacillus salivarius subsp. salicinius로 동정되었다.

• 한국축산식품학회지
• /
• 제27권1호
• /
• pp.102-109
• /
• 2007

Lactobacillus salivarius subsp. salivarius CNU27 possessed a high level of ${\alpha}$-galactosidase activity. Purified ${\alpha}$-galactosidase was obtained after sonication of harvested cell pellet followed by DEAE-Sephadex A-50 and Mono Q anion exchange chromatography. The specific activity of the purified enzyme was 8,994 units/mg protein which is 17.09 times higher than that in crude extract. The native enzyme was a monomer with a molecular mass of 56,397.1 dalton. The optimum temperature and pH for the enzyme were $40^{\circ}C$ and 6.0, respectively. The enzyme was stable between 25 and $50^{\circ}C$. However, ${\alpha}$-galactosidase activity was lost rapidly below pH 4.5 and above pH 8.5. The enzyme activity decreased to 6.73% and 4.30% of the original activity by addition of $Cu^{2+}$ and $Hg^{2+}$, respectively. Other metal compounds did not affect the enzyme activity significantly. The enzyme liberated galactose from melibiose, raffinose, and stachyose. The rate of substrates hydrolysis was measured by HPLC. Raffinose, stachyose and melibiose were completely decomposed after 24 hr at $40^{\circ}C$.

• 한국축산식품학회지
• /
• 제27권1호
• /
• pp.110-116
• /
• 2007

Lactobacillus salivarius subsp. salivarius Nam27 with a high ${\beta}$-galactosidase activity was selected for enzymatic characterization. For purification, cell pellet was disrupted by Bead Beater, by DEAE-Sepharose and Mono-Q chromatography. The specific activity of the purified enzyme was 5,312 units/mg. The molecular weight of native monomeric ${\beta}$-galactosidase was estimated to be 30,000 dalton (monomer) by the SDS-PAGE. The optimum temperature and optimum pH were $50^{\circ}C$ and 5.0, respectively. This enzyme was stable between 35 and $55^{\circ}C$. ${\beta}$-galactosidase activity was lost rapidly above pH 7.0. But ${\beta}$-galactosidase was more stable at pH 4.0 (acidic conditions). And ${\beta}$-galactosidase activity was lost rapidly above $65^{\circ}C$ after 10 min incubation. $Ca^{2+}$ and $Zn^{2+}$ metal ions enhanced ${\beta}$-galactosidase activity by 164.09% and 127.37% while $Cu^{2+}$, $Fe^{3+}$ and $Mn^{2+}$ lowered ${\beta}$-galactosidase activity by 58.29%,85.10% and 77.66% respectively. Other metal ions didn't affect ${\beta}$-galactosidase activity significantly.

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

• 한국치위생학회지
• /
• 제5권2호
• /
• pp.131-145
• /
• 2005

There are normal inhabitants doing medically useful functions in the body. There are many kinds of bacteria performing specific functions in the oral cavity. Two strains of lactic add bacteria were isolated from normal inhabitants of children's oral cavity, which inhibited the production of volatile sulfur compounds by anaerobic bacteria. The authors identified the isolates by 16S rDNA partial sequencing. 1. Two isolates were Gram-positive bacilli and produced hydrogen peroxide. 2. When Streptococcus mutans was cultured in the media, the mean weight of formed artificial plaque on the orthodontic wires was $124.4{\pm}30.4$ mg, whereas being reduced to $5.2{\pm}2.0$ mg and $10.6{\pm}6.6$ mg in the media cultured with Streptococcus mutans and each isolate, respectively(p<0.05). 3. The number of viable cells of Streptococcus mutans was $3.4{\times}10^9$ per ml in the cultured solution, whereas those of Streptococcus mutans in the combined culture with each of isolates were $4.6{\times}10^8$ and $2.4{\times}10^8$ per ml. 4. The optical density was 1.286 in the supernatant of Fusoacterium nucleatum after vortexing for 30 minutes, whereas in the supernatant of combined Fusoacterium nucleatum and each isolate, they were reduced to 0.628 and 00497, which the percentages of coaggregation between them were 2904% and 57.8%, respectively. 5. The optical density of Fusoacterium nucleatum precipitate was 1.794 in the culture media containing cysteine and $FeSO_4$ being reduced to 1.144 and 0.915 in the coaggregated precipitates of Fusoacterium nucleatum and each isolate. 6. The similarity values of 16S rDNA sequence between each of isolates and Lactobacillus salivarius subsp. salicinius were 99.60% and 99.73%, respectively, meaning that isolates were Lactobacillus salivarius subsp. salicinius. These results indicated that two strains isolated from children's saliva, which inhibited the formation of plaque and the production of volatile sulfur compounds, were identified as Lactobacillus salivarius subsp. salicinius.