• Title/Summary/Keyword: Lactase

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Cloning and Expression of Kluyveromyces fragilis $\beta$-Galactosidase Gene in Saccharomyces cerevisiae

  • Bang, Jeong-Hee;Nam, Doo-H.;Kang, Dae-Ook;Ahn, Jong-Seog;Ryu, Dewey-D.Y.
    • Journal of Microbiology and Biotechnology
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    • v.5 no.1
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    • pp.6-13
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    • 1995
  • A gene coding for the $\beta$-galactosidase (lactase) of Kluyveromyces tragilis UCD 55-55 was isolated by complementation in Escherichia coli YMC9. From the plasmid library made from Sau3A-digested chromosomal DNA, one positive clone was selected. The cloned gene for $\beta$-galactosidase was on 7.3 kilobase pair DNA fragment, and a slightly low level of $\beta$-galactosidase enzyme activity was detecied in E. coli. It was also confirmed that the cloned gene comes from K. tragilis by DNA-DNA hybridization and immunochemical blotting experiments. In order to construct a new yeast strain having the metabolic ability for lactose, the cloned gene for K. tragilis $\beta$-galactosidase was inserted in yeast vector YEp24 and YRp17, and transformed into Saccharomyces cerevisiae YNN27 and Ml-2B. The yeast transformants showed the nearly the same $\beta$-galactosidase productivity as level of K. tragilis when uninduced, but these could not utilize lactose as a sole carbon source, presumably due to the lack of lactose transport system. Nevertheless, a slightly higher ethanol productivity was achieved by these transformants than S. cerevisiae or K. tragilis, in the medium containing glucose and lactose.

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Hypoglycemic Effect of the Giant Embryonic Rice Supplementation on Streptozotocin-induced Diabetic Rats (거대배아미 식이에 의한 Streptozotocin 유발 당뇨 흰쥐의 혈당강하 효과)

  • Lee, Youn-Ri;Nam, Seok-Hyun;Kang, Mi-Young
    • Korean Journal of Food Science and Technology
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    • v.38 no.3
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    • pp.427-431
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    • 2006
  • This study was performed to investigate the influence of giant embryo feeding on hypoglycemia in streptozotocin-induced diabetic rats. Streptozotocin -induced diabetic rats were fed four kinds of experimental diets such as corn starch diet as a control, polished rice diet, brown rice diet and giant embeyonic rice diet for 6 weeks. The body weight gain diabetic of control was signaficantly suppressed compared to that of the normal. The supplementation of giant embryo increased the surivival rate. Feeding of giant embryo appeared to have ameliorating effects on diabetic symptoms including features such as polyphasia, polyuria. Giant embryo shortened gastrointestinal transit time and increased total fecal weight, total fecal dry weight and fecal water content compared with diabetic control. Giant embryo showed fasting blood glucose lowering effect compared with diabetic control. The disaccharidase activities in proximal part of intestine such as maltase sucrase and lactase in giant embryo feeding groups were lower than diabetic control. The results of this study show that giant embryo supplementation may have a beneficial Veffect on the hypoglycemia may be useful in the diet therapy for diabetic.

Oral Insulin-like Growth Factor-I Combined Alters Intestinal Protein Synthesis in Parenterally-fed Piglets

  • Park, Yoo-Kyoung;Sharon M. Donovan
    • Nutritional Sciences
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    • v.3 no.2
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    • pp.57-65
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    • 2000
  • Partial enteral nutrition (PEN) supplemented with insulin-like growth factor-I (IGF-I) to neonatal piglets receiving parenteral nutrition increases lactase-phlorizin hydrolase (LPH) activity, but not LPH mRNA. The goal of the current study was to investigate the mechanism by which IGF-I up-regulates LPH activity. We hypothesized that IGF-I regulates LPH synthesis post-transcriptionally. Methods: Newborn piglets (n=15) received 100% parenteral nutrition (TPN), 80% parenteral nutrition + 20% PEN (PEN), or PEN + IGF-I (1.0mg/kg/d). On day 7, two stable isotopes of leucine, [$^2 H_3$]-leucine and [$^{13}C_1$]-L-leucine were intravenously administered to measure mucosal protein and brush LPH (BB LPH) synthesis. Results: Weight gain, nutrient intake and jejunal weight and length were similar among the treatment groups. PEN increased mucosal weight, villus width and cross-sectional area, LPH activity, mRNA expression and the abundance of proLPHh compared to 100% TPN (p<0.05). IGF-I further increased mucosal weight, LPH activity and LPH activity per unit BB LPH ~2-fold over PEN alone (p<0.05), but did not affect LPH mRNA or the abundance of proLPHh or mature LPH. Isotopic enrichment of [$^2 H_3$]-leucine and [$^{13}C_1$]-L-leucine in plasma, mucosal protein and LPH precursors, and the fractional and absolute synthesis rates of mucosal protein and LPH were similar among the treatment groups. Total mucosal protein synthesis was increased 60% (p<0.05) and LPH synthesis tended (p=0.14) to be greater in the IGF-I treated animals compared to the other two groups. Conclusions: The primary mechanism by which IGF-I up-regulates LPH may be post-translational, either via reducing LPH turnover, or by specifically altering LPH activity.

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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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Change of Fatty Acid in Cheese Ripening by New Development of Lactic Acid Bacteria (육종 균주에 의한 치즈의 지방산 성분 변화에 관한 연구)

  • 송재철;김정순;박현정;신완철
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.6
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    • pp.1068-1076
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    • 1997
  • This study was carried out to elucidate the utility of fusant for shortening the ripening time of imitation processed cheese. L. bulgaricus exhibited the highest protease and lactase activity and L. helveticus revealed the highest lipase activity. Fusant was shown to be high in the activity of protease and lactase. The total volatile free fatty acid produced by the cheese treated with L. helveticus was markedly increased after four ripening days and was gently increased after nine ripening days. However, L. bulgaricus significantly increased the total volatile free fatty acid between four and nine ripening days. In the case of fusant, the amount of total volatile free fatty acid was observed to increase at a constant rate relative to the ripening time. In free fatty acid production at different ripening times, L. bulfaricus generated caproic acid and caprilic acid abundantly while it produced a poor quantity of capric acid, lauric acid, and myristic acid. In the cheese sample treated with L. helveticus, the amount of caproic acid and capylic acid was on increase as the ripening time increased. The amount of caproic acid and caprylic acid produced by fusant was less than that produced by the other two starters. In the panel sensory evaluation, the flavor intensity and preference increased as the ripening time increased. The cheese sample treated with fusant showed the highest flavor intensity at 7 days, whereas cheese treated with L. helveticus exhibited the highest flavor intensity at 15 or 30 days. The cheese treated with L. helveticus showed the highest preference at 7 days, but cheese treated with fusant exhibited the highest preference at 30 days.

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Effect of Sorbitol on the Texture and the Survival of Lactic Acid Bacteria of Frozen Yoghurt (Sorbitol첨가가 Frozen Yoghurt중 유산균의 생존율과 조직에 미치는 영향)

  • Kim, Eung-Ryul;Kim, Yu-Seong;Lee, Yeong-Geon;Ju, Ji-Seon;Lee, Gang-Ik;Baick, Seung-Cheon;Lee, Jong-Ik;Yu, Je-Hyeon
    • Journal of Dairy Science and Biotechnology
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    • v.14 no.1
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    • pp.71-84
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    • 1996
  • This study was conducted to examine the usability of sorbitol for the manufacture as low-calory ingredient and cryoprotectant against frost damage. When frozen yoghrt was made of replacing sucrose by sorbitol at yoghurt mix, the change of physicochemical and lactic acid bacteria, such as Str. thermophilus, L. bulgaricus, and mixed culture of Str. thermophilus, and L. bulgaricus(1:1), was studied during the frozen storage(-20$^{\circ}$C). During the incubation of yoghurt mix, the rapid growth of lactic acid bacteria in all sample was observed as the increase of sorbitol addition, but sample A and D were almost similar. This results suggested that sucrose could play role of effecting the growth stimulator, otherwise, sorbitol could inhibit the death of microorganism, following the genus. At the survival rate between lactic acid bacteria during freezing of -5$^{\circ}$C by ice cream freezer Str. thermophilus showed 26.19 to 34.76%, L. bulgaricus 3.97 to 5.20%, and mixed culture 17.16 to 40.87% respectively. L. bulgaricus showed the greater lethal rate than other genus. Sample C which mixed sucrose with sorbitol (1:2 ratio) was showed the lowest lethal rate. Therefore, it suggested that the use of this ration could be used for better anti-frost damage. During the storage of -20$^{\circ}$C, the number of lactic acid bacteria generally decreased in the stand point of genus and frozen storage period. The survival of lactic acid bacteria might be the addition of sorbitol which could have the effect of anti-forst damage. In all treatment, lactase activity showed the rapid decrease after freezing. During the period of frozen storage, it was shown the slow decreasing trend. In spite· of decreasing, the result during yoghurt mix incubation -5$^{\circ}$C freezing, and -20$^{\circ}$C frozen storage was different at the level. After 80 days of storage, the lactase activity was similar among all genus and sample. Despite differenting viscosity followed by genus, combination of mix, and pH, the ratio of 1 to 2(sucrose : sorbitol) showed the greatest viscosity. The water holding capacity of frozen yoghurt mix was closely related to viscosity. As increasing sorbitol amounts, hardness and cohesiveness were increased, but elastisity was decreased. The significant differences between sample was inoculated with Str. thermophilus. However, there were not significant difference from the sample inoculated with L. bulgaricus and mixed culture.

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Identification of Novel Psychrotolerant Bacterial Strain and Production of $\beta-Galactosidase$ (새로운 저온 내성세균의 동정과 $\beta-Galactosidase$ 생산)

  • Park, Jeong-Woon;Yoo, Jae-Soo;Roh, Dong-Hyun
    • Korean Journal of Microbiology
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    • v.42 no.1
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    • pp.40-46
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    • 2006
  • Galactose joined to glucose by a $\beta(1\rightarrow4)$ glycosidic bond makes lactose and this disaccharide is rich in milk. It is known that lacotse is hydrolyzed to each monomeric sugar by either lactase in human or $\beta-galactosidase$ in bacteria. Ingestion of milk by lactase-deficient persons causes a temporary diarrhea and subsequent chronic diarrhea results in colitis with chronic inflammation. We isolated a $\beta-galactosidase$ producing psycrotolerant strain AS-20 from near cattle shed and investigated the growth at various temperature conditions. Whereas Escherichia coli strains did not grow at $10^{\circ}C$, the AS-20 strain could grow well at this low temperature and showed optimal growth at $30^{\circ}C$. The isolated strain was identified as 97% Hafnia alvei by biochemical properties. This strain could ferment glucose, lacotse, maltose, mannitol, xylose, ONPG, rhamanose and L-arabinose, and decarboxylate lysin and ornithine. To confirm the identity of isolated strain we amplified 16S rDNA by PCR and searched similarity of the 1426 bp DNA sequcence with Genbank database. The strain AS-20 showed 99% similarity with Hafnia alvei. The activity of $\beta-galactosidase$ was 1.5 times higher when the cell was grown at 10 or $20^{\circ}C$ than at $30^{\circ}C$. The highest enzyme activity of AS-20 was also much higher than that of E. coli, which was grown at $30^{\circ}C$.

Blood Glucose Lowering Activity and Mechanism of Supungsungihyan (SPSGH) in db/db Mouse (db/db 마우스에서 수풍순기환의 혈당강하 활성 및 기전연구)

  • 이성현;안세영;두호경;정성현
    • Biomolecules & Therapeutics
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    • v.7 no.4
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    • pp.335-341
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    • 1999
  • Antidiabetic activity and mechanism of Supungsungihyan(SPSGH) were examined in db/db mice, which is a spontaneously hyperglycemic, hyperinsulinemic and obese animal model. SPSGH and acarbose were administered orally for 4 weeks. Fasting and non-fasting serum glucose, glycated hemoglobin and trig-lyceride of SPSGH treated group were all reduced when compared with those of db/db control group. At 12th week after birth, SPSGH increased an insulin secretion although statistic significance was not seen. Total activities of sucrose, maltase and lactase in SPSGH treated group were not significantly different from those in db/db control. On the other hand, sucrase and maltase activities in acarbose treated groups were increased. Effect of SPSGH on mRNA expression of glucose transporter(GLUT-4) was also examined by RT-PCR and in vitro transcription with co-amplification of rat $\beta$-actin gene as an internal standard. Muscular GLUT-4 mRNA expression in SPSGH treated group was increased significantly. These results may suggest that SPSGH lowered blood glucose ascribing to upregulation of muscular GLUT-4 mRNA expression.

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Nanofood Materials and Approachable Development of Nanofunctional Dairy Products (나도 식품 소재와 나노 기능성 유제품 개발의 가능성)

  • Gwak, Hae-Su;Kim, Dong-Myeong
    • Journal of Dairy Science and Biotechnology
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    • v.22 no.1
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    • pp.1-12
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    • 2004
  • Nanofood is advanced functional food which food industry and food scientist try to develop process foods in near future. To be developed nanofood, nanofood materials are needed, such as biodegradable nanosphere material, biotechnical nanofood material, and protein and nanofood material. There are some food industrial applications with nanotechnology, such as nanoencapsulation, nanomolecule making, nanoparticle and powder making, nano separation, and nano extration. We can find several nanofoods and nanofood materials on the market. In addition, dairy industry is also in the first step for the development of nanofunctional food. However, nanoencapsulations of lactase, iron, vitamin C, isoflavone are developed for functional milk. Dairy industry needs various nanofood materials to be advanced functional dairy products.

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Effect of Silkworm Extract on Intestinal $\alpha$-Glycosidase Activity in Mice Administered with a High Carbohydrate-containing Diet (고탄수화물 식이투여 마우스에서 누에추출물이 소장내의 $\alpha$-glycosidase 활성에 미치는 영향)

  • Jeong, Seong-Hyeon;Kim, Mi-Seon;Ryu, Gang-Seon
    • Journal of Sericultural and Entomological Science
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    • v.39 no.1
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    • pp.86-92
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    • 1997
  • The effects of silkworm extract on body weight, blood glucose and insulin levels, and an intestinal glycosidase activity were studied in high carbohydrate diet administered mice. Hyperglycemia and hyperinsulinemia represented in high carbohydrate diet administered mice disappeared when silkworm extract(50 mg) was added to 100 g diet for 10 week period. These results suggest that silkworm extract may resolve the insulin resistance by lowering serum insulin level resulting from inhibition of intestinal glycosidase activity. Silkworm extract coadministered with high carbohydrate diet to mice for 10 weeks significantly induced maltase, sucrase and lactase acivities, especially middle and distal portion. Further studies about regulatory mechanisms of glycosidase by silkworm extract are needed.

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