• Title/Summary/Keyword: $\beta$-Lactose

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Mapping of Gene Encoding Phospho-$\beta$-galactosidase from Lactobacillus casei and its Expression in Escherichea coli (Lactobacillus casei 의 Phospho-$\beta$-galactosidase 유전자의 지도작성과 Escherichia coli 내에서의 발현)

  • 박정희;문경희;민경희
    • Korean Journal of Microbiology
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    • v.30 no.6
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    • pp.539-545
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    • 1992
  • Recombinant plasmid pPLac15 determined both phosphoenolpyruvate-dependent phosphotransferase uptake of lactose and phospho-$\beta$-galactosidase (Moon et al., 1989). A restriction mapping of the pPLac15 was compiled with several restriction enzymes and a seriese of sub clones into pUC18 was constructed. From an analysis of the proteins produced by Escherichia coli cells of transformants containing each of the recombinant subclone plasmids, it was found that the gene for phospho-$\beta$-galactosidase in pUCI8 was expressed about 1.8-folds in E. coli.

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${\beta}-galactosidase$ Activity of Lactobacillus spp. from Pickles (젓갈에서 분리한 Lactobacillus spp.의 ${\beta}-galactosidase$ 특성)

  • Rhee, Young-Hwan;Oh, Min-Keun;Lee, Yong-Kyu;Shin, Seung-Yee;Kim, Jong-Hyun
    • Applied Biological Chemistry
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    • v.39 no.6
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    • pp.437-442
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    • 1996
  • Two strains of Lactobacillus(L.) casfi and one strain of L. Pentosus, which were isolated from pickles, were used to investigate in studing their characteristics of ${\beta}-galactosidase$. The preferable carbon sources and pH of the MRS media for enzyme production from L. casei No.10 was found to be 1.0% lactose and pH 7.5, from L. Pentosus No.63 was 1.0% galactose and pH 7.5, and from L. casei No.36 was 1.0% lactose and pH 6.5, respectively. The maximum enzyme production from each strain was found after 48 hours culture at $30^{\circ}C$ in a medium with preferable carbon source. The optimum reaction temperature with substrate for ${\beta}-galactosidase$ activity was found at $60^{\circ}C$ for all three strains . The stability of enzyme from L. casei No.36 was found to be at $45^{\circ}C$, from L. Pentosus No.63 was found at $55^{\circ}C$. This stability from L. casei No.36 was found at $40^{\circ}C$, but it was reduced to 60% at $55^{\circ}C$. These stabilities of enzymes remained about 90% at $40^{\circ}C$ for all three strains. The optimal pH for enzyme activities was found to be pH 6.5 for all three strains. Enzyme activity remained over 90% for L. casei No.10 at $pH\;5.0{\sim}6.0$, for L. casei No.36 at $pH\;5.0{\sim}8.0$, and for L. pentosus No.63 at $pH\;6.0{\sim}7.0$.

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Fed-batch Fermentations of Recombinant Escherichia coli to Produce Bacillus macerans CGTase

  • Park, Yong-Cheol;Kim, Chang-Sup;Kim, Chung-Im;Choi, Kyu-Hwan;Seo, Jin-Ho
    • Journal of Microbiology and Biotechnology
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    • v.7 no.5
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    • pp.323-328
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    • 1997
  • The recombinant Escherichia coli BL21(DE3)pLysE : pTCGT1 was grown to overproduce Bacillus macerans cyclodextrin glucanotransferase (CGTase) able to synthesize ${\alpha}$-cyclodextrin (CD) with a selectivity of 67%. A number of batch fermentations were performed to test the possibility of using lactose as an inducer of the E. coli T7 promoter system. A mixture of isopropyl ${\beta}$-D-thiogalactoside (IPTG) and lactose (1 : 1) gave a maximum CGTase activity of 2.4 U/ml, which was higher than the value obtained with induction by IPTG alone. Fed-batch fermentations involving a glucose-controlled growth period followed by a gene-expression phase with mixtures of IPTG and lactose were employed to achieve high cell density and thereby increase total CGTase activity. Optimized fed-batch fermentation using the modified inducer (IPTG : lactose=1 : 3) and 100 g/l yeast extract solution in the gene-expression phase resulted in a maximum CGTase activity of 62.9 U/ml and a final cell mass of 53.5 g/l, corresponding to a 31-fold increase in CGTase activity and a 29-fold increase in cell mass compared with the control batch fermentation.

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Effect of Milk Containing Streptococcus thermophilus KACC 91147 on Blood Glucose Levels (Streptococcus thermophilus KACC 91147 첨가우유 섭취가 혈당치에 미치는 영향)

  • Ham, Jun-Sang;Jeong, Seok-Geun;Noh, Young-Bae;Shin, Ji-Hye;Han, Gi-Sung;Chae, Hyun-Seok;Yoo, Young-Mo;Ahn, Jong-Nam;Cho, Yong-Min;Kim, Geun-Bae
    • Food Science of Animal Resources
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    • v.27 no.4
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    • pp.496-500
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    • 2007
  • The lactase activities of nine species of lactic acid bacteria were compared using the chromogenic substrate, $o-nitrophenyl-{\beta}-D-galactopyranoside$. Streptococcus thermophilus KACC 91147 had the highest lactase activity among a total of thirty strains of Lactobacillus and S. thermophilus tested, including commercial strains. S. thermophilus KACC 91147 released $0.30{\pm}0.12\;mg/mL$ of galactose in treated milk A ($10^7\;CFU/mL$) and $6.49{\pm}0.38\;mg/mL$ in treated milk B ($10^9\;CFU/mL$ milk) over 2 hours. In milk tolerance tests, the blood glucose level (BGL) of 6 volunteers (2 males and 4 females) clinically diagnosed as lactose intolerant increased 3.0 mg/dl after drinking milk A, but a significant (p<0.05) additional increase of $11.2{\pm}4.18\;mg/dl$ was found after drinking milk B. This result suggests that the addition of S. thermophilus KACC 91147 cells into milk aids the digestion of lactose in milk and ameliorates the symptoms of lactose-intolerant individuals due to the activity of lactase from the lactic streptococci.

Studies on the Production of $\beta$-Galactosidase by Lactobacillus sporogenes - Production of Extracellular $\beta$- Galactosidase - (Lectobacillus sporogenes에 의한 $\beta$-Galactosidase 생산에 관한 연구 ( I ) -균체외 $\beta$-Galactosidase의 생산 -)

  • 김영만;이정치;정필근;최용진;양한철
    • Microbiology and Biotechnology Letters
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    • v.11 no.1
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    • pp.59-66
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    • 1983
  • Cultural conditions for the production of extracellar $\beta$-galactosidase by Loctobacillus sporogenes, a spore forming lactic acid bacterium, were investigated with shaken flask and jar fermenter cultures. The fermentation medium giving maximum $\beta$-galactosidase yield was found to consist of 1 % lactose as a carbon source, 1.5% peptone as an organic nitrogen source. 0.2% ammonium sulfate as an inorganic nitrogen source, 0.8% ammonium phosphate dibasic as a phosphorus source, and 0.05% potassium chloride and 0.001% ferric chloride as mineral source. Optimal initial pH of the medium was 7.0 and the highest enzyme excretion was observed after 40 hours of cultivation at 37$^{\circ}C$. In this experiment, the 500$m\ell$ conical flask containing 50-200$m\ell$ of medium was shaken at 140 strokes per minute with 7cm amplitude in a reciprocating shaker. The maximum enzyme value attained was 38 U/$m\ell$ of the culture broth which was found to be slightly higher than the highest intermolecular enzyme activity (30 U/$m\ell$) observed after 24 hours of incubation. In the fermentor culture, the fermentation profile was shown to be similar to that observed in the shaken flask experiment. But the maximum extracellular enzyme activity was 45 U/$m\ell$ to be even higher than the value obtained with the shaken flask culture.

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Characterization of a potential probiotic strain Lactobacillus plantarum MY4

  • Jeong, Hwang-Yeong;Jeong, Min-Yong;Kim, Gi-Tae;Kim, Cheon-Je;Baek, Hyeon-Dong
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.882-885
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    • 2001
  • The cells of Lactobacillus plantarum MY4 isolated from the human feces were treated for 24 h in artificial bile after incubation for 2 h in artificial gastric juice and final number of the strain was reached to around $3.1{\times}10^8$ CFU/ml. In test of API ZYM kit, ${\beta}$-glucuronidase or ${\beta}$-glucosidase were not produced by L. plantarum MY4. However, ${\beta}$-galactosidase were weakly produced by it, which they would be alleviated the lactose intolerance. L. plantarum MY4 were resistant to antibiotics such as nisin, tetracycline, streptomycin, rifamycin, doxycycline, roxithromycin, chloramphenicol, nystatin, erythromycin, ciprofloxacin and gentamycin. L. plantarum MY4 was affected by alcohol concentration up to 8%, but more than 16%, their growth was not affected significantly. L. plantarum MY4 was shown to inhibit the growth of Listeria monocytogenes ATCC 19111 completely within 24 h of incubation, which indicates its bactericidal nature. Thus, L plantarum MY4 show promise as a probiotic strain because of its characteristics.

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새로운 오메프라졸염의 개발 및 제제화 연구

  • 이계주;이기명;이창현;우종수
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.04a
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    • pp.329-329
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    • 1994
  • OMZ과 HP-$\beta$-CD 포접화합물의 유형은 $A_{L}$ type의 가용성 복합체이며, CD, UV, IR, DSC, XRD pattern 및 $^1$H-NMR 방법으로 측정한 포접화합물의 결함형태는 OMZ의 benzimidazole부분이 HP-$\beta$-CD 공동에 포접된 형태이었고, OMZ과 HP-$\beta$-CD의 결합조성몰비는 1:1, 안정도 상수는 약 34 M$^{-1}$이었다. 또한 cholestyramine 수지와 OMZ 복합체는 방출양상이 신속하였다. OMZ-cholestyramine 수지염은 안정성이 양호하고 용출율이 우수한복합체로 경구용 재제를 개발할 수 있었으며, 용출시험 및 산 저항성이 양호하고 pellet의 제조에 있어서 core의 제조는 정제수 : 에탄올(7: 3) 결합액을 이용하여 lactose를 주로 사용하고 기타 PEG 6000, sorbitol, Avicel PH101, sodium lauryl sulfate 및 무수 PVP K-30을 적절히 배합하여 제조하고, 장용코팅의 내산성은 90.5% 이었으며, 소장에서의 용출경향이 85%이상이 10분 이내에 용출되어 규정에 적합하였다.

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Effect of Antioxidant Supplementation in Freezing Extender on Porcine Sperm Viability, Motility and Reactive Oxygen Species

  • Park, Sang-Hyoun;Yu, Il-Jeoung
    • Journal of Embryo Transfer
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    • v.32 no.1
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    • pp.9-15
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    • 2017
  • The present study was aimed to determine the effect of green tea extract (GTE) and beta-mercaptoethanol (${\beta}$-ME) supplementation in boar sperm freezing extender on sperm motility, viability and reactive oxygen species (ROS) level. Experimental groups were allocated into Lactose-egg yolk (LEY) without antioxidant (control), GTE (1,000 mg/L GTE in LEY) and ${\beta}$-ME ($50{\mu}M$ ${\beta}$-ME in LEY). Spermatozoa extended with LEY were cooled to $5^{\circ}C$ for 3 h and then kept at $5^{\circ}C$ for 30 min following dilution with LEY containing 9% glycerol and 1.5% Equex STM (final sperm concentration: $1{\times}10^8/mL$). Spermatozoa were loaded into straws and frozen in nitrogen vapor for 20 min. Following thawing at $37^{\circ}C$ for 25 sec, sperm viability and ROS level were measured using fluorescent double stain Fertility(R) and cytometry, respectively. Motility and viability of GTE supplemented-group were higher than those of control and ${\beta}$-ME without significance. ROS level in GTE group showed significantly lower than control (P < 0.05). In conclusion, GTE supplementation in boar sperm freezing extender can reduce ROS generation during freezing.

The effect of carrot juice, ${\beta}$-carotene supplementation on lymphocyte DNA damage, erythrocyte antioxidant enzymes and plasma lipid profiles in Korean smoker

  • Lee, Hye-Jin;Park, Yoo-Kyoung;Kang, Myung-Hee
    • Nutrition Research and Practice
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    • v.5 no.6
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    • pp.540-547
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    • 2011
  • High consumption of fruits and vegetables has been suggested to provide some protection to smokers who are exposed to an increased risk of numerous cancers and other degenerative diseases. Carrot is the most important source of dietary ${\beta}$-carotene. Therefore, the objective of this study was to investigate whether carrot juice supplementation to smokers can protect against lymphocyte DNA damage and to compare the effect of supplementationof capsules containing purified ${\beta}$-carotene or a placebo (simple lactose). The study was conducted in a randomized and placebo-controlled design. After a depletion period of 14 days, 48 smokers were supplemented with either carrot juice (n = 18), purified ${\beta}$-carotene (n = 16) or placebo (n = 14). Each group was supplemented for 8 weeks with approximately 20.49 mg of ${\beta}$-carotene/day and 1.2 mg of vitamin C/day, as carrot juice (300 ml/day) or purified ${\beta}$-carotene (20.49 mg of ${\beta}$-carotene, 1 capsule/day). Lymphocyte DNA damage was determined using the COMET assay under alkaline conditions and damage was quantified by measuring tail moment (TM), tail length (TL), and% DNA in the tail. Lymphocyte DNA damage was significantly decreased in the carrot juice group in all three measurements. The group that received purified ${\beta}$-carotene also showed a significant decrease in lymphocyte DNA damage in all three measurements. However, no significant changes in DNA damage was observed for the placebo group except TM (P = 0.016). Erythrocyte antioxidant enzyme was not significantly changed after supplementation. Similarly plasma lipid profiles were not different after carrot juice, ${\beta}$-carotene and placebo supplementation. These results suggest that while the placebo group failed to show any protective effect, carrot juice containing beta-carotene or purified ${\beta}$-carotene itself had great antioxidative potential in preventing damage to lymphocyte DNA in smokers.

Lactase activity in yoghurt and lactic acid bacteria (요구르트와 유산균에서의 Lactase Activity)

  • Lee, Kwang-Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.21 no.1
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    • pp.60-63
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    • 1992
  • Individual starter culture were inoculated into liquid medium and incubated at $40^{\circ}C$ for 16 hours. Whole cell were obtained and evaluated for ${\beta}-galactosidase$ activity using orthonitrophenyl-${\beta}-D-galactopyranoside$ (ONPG) as substrate. S. thermophilus had more ${\beta}-galactosidase$ activity than other Lactobacilli did. To study the effect of storage temprature on enzyme activity of yoghurt, some samples of cultured yoghurt were stored under refrigeration $(4^{\circ}C)$, and the others under room temperature $(23^{\circ}C)$. At $4^{\circ}C$, yoghurt had ${\beta}-galactosidase$ activity and many viable bacteria in 1 month. After 20 days, yoghurt had maximum ${\beta}-galactosidase$ activity. At $23^{\circ}C$, yoghurt had ${\beta}-galactosidase$ activity by 5 days. As this experiment shown ${\beta}-galactosidase$ activity was ascribed to viable bacteria, especially S. thermophillus. Commercial yoghurt had lower ${\beta}-galactosidase$ activity. There were considerable variations with regard to the lactose hydrolyzing capabilities of commercial yoghurt samples.

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