• Title/Summary/Keyword: /alpha-galactosidase

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Detection of Chlorotoluene and Nitrotoluene Compounds by Recombinant Microbial Biosensors (재조합 미생물 바이오센서를 이용한 chlorotoluene과 nitrotoluene 화합물의 검출)

  • Lee, Da Young;Cho, Jae Ho;Lim, Woon Ki;Shin, Hae Ja
    • Journal of Life Science
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    • v.24 no.1
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    • pp.54-60
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    • 2014
  • Aromatic hydrocarbons are toxic environmental pollutants that are detrimental to the ecosystem and human health. Among them, chlorotoluene and nitrotoluene are toxic to hydrobios and irritate the skin, eyes, and respiratory organs of humans. We herein report the development of recombinant microbial biosensors for cheap and rapid monitoring of chlorotoluene and nitrotoluene compounds. Plasmids were constructed by inserting the xylR regulatory gene for BTEX (benzene, toluene, ethylbenzene, and xylene) degradation into upstream of Po' (the DmpR activator promoter Po with the deletion of its own upstream activating sequences) or Pu (the cognate promoter of XylR)::lacZ (the ${\beta}$-galactosidase gene) and transformed into Escherichia coli $DH5{\alpha}$. In the presence of inducers, the biosensor cells immobilized in agarose developed a red color in 1-2 h due to the hydrolysis of chlorophenol red ${\beta}$-D-galactopyranoside (CPRG), a substrate of ${\beta}$-galactosidase that was expressed by the inducers. Among BTEX, high responses were specifically observed with o-, m-, p-chlorotoluene ($0.1{\mu}M-100 mM$) and o-, m-, p-nitrotoluene (0.1 mM-100 mM). Po' demonstrated higher responses than those with Pu. The biosensors immobilized in agarose showed good stability after 21 days' storage at $4^{\circ}C$, and responses in untreated wastewater spiked with chlorotoluene and nitrotoluene, suggesting they can be used to detect compounds in wastewater.

Physiological Characteristics and Anti-Obesity Effect of Lactobacillus plantarum K6 isolated from Kimchi (김치에서 분리한 Lactobacillus plantarum K6의 생리적 특성 및 비만억제효과)

  • Kim, Seulki;Lim, Sang-Dong
    • Journal of Dairy Science and Biotechnology
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    • v.35 no.4
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    • pp.221-231
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    • 2017
  • This study aimed to investigate the physiological characteristics and anti-obesity effects of a newly isolated bacterium, Lactobacillus plantarum K6. L. plantarum K6 showed good ${\alpha}-amylase$ inhibitory activity ($96.78{\pm}3.29%$), ${\alpha}-glucosidase$ inhibitory activity ($92.55{\pm}9.62%$), and lipase inhibitory activity ($85.17{\pm}0.79%$), and the strain inhibited the adipocyte differentiation of 3T3-L1 cells ($27.4{\pm}1.4%$) when present at a concentration of $100{\mu}g/mL$. L. plantarum K6 was isolated from kimchi and its physiological characteristics were investigated. A comparison of the sensitivity of the isolate to 15 different antibiotics showed that L. plantarum K6 is highly sensitive to erythromycin and highly resistant to vancomycin, ampicillin, and polymyxin B. This strain also showed high arylamidase and ${\beta}-galactosidase$ activities. Moreover, it was relatively tolerant to bile acid and low pH, and displayed resistance to Escherichia coli, Salmonella Typhimurium, Listeria monocytogenes, and Staphylococcus aureus, with rates of 51.8%, 42.4%, 61.6%, and 54.9%, respectively. No bio genic amines were produced. L. plantarum K6 also showed high adhesion activity to HT-29 cells compared to L. rhamnosus GG. These results demonstrate that Lactobacillus plantarum K6 has potential as a probiotic with anti-obesity effects.

Pectolytic Enzymes of the Industrial Fungus Aspergillus kawachii

  • Vita, Carolina Elena;Esquivel, Juan Carlos Contreras;Voget, Claudio Enrique
    • Food Science and Biotechnology
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    • v.18 no.6
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    • pp.1365-1370
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    • 2009
  • Aspergillus kawachii extracellular pectinases were screened in liquid cultures with different carbon sources. The fungus grown on citrus pectin or lemon pomace produced at least one of these inducible pectinases: acidic polygalacturonase, pectin lyase, pectin methylesterase, $\alpha$-L-arabinofuranosidase, $\alpha$-1,5-endoarabinase, $\beta$-D-galactosidase/exogalactanase, and $\beta$-1,4-endogalactanase. The lemon-pomace filtrates also contained significant $\alpha$-L-rhamnosidase and $\beta$-D-fucosidase activities. Most of the screened pectinases were active at pH 2.0-2.5, indicating that the A. kawachii enzymes were acidophilic. Under the culture conditions employed we could not detect enzymatic degradation of soybean rhamnogalacturonan. The A. kawachii pectinase-production-related regulatory phenomena of induction-repression resemble those described for other Aspergillus sp.

Effects of Alpha-galactosidase Supplementation to Corn-soybean Meal Diets on Nutrient Utilization, Performance, Serum Indices and Organ Weight in Broilers

  • Wang, C.L.;Lu, W.Q.;Li, Defa;Xing, J.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.12
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    • pp.1761-1768
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    • 2005
  • Effects of alpha-galactosidase (GAL) on broiler corn-soybean meal diet was investigated. In experiment 1, sixty cockerels were allocated to five groups, including three enzyme treatments (GAL added at 0, 500, and 1,000 mg/kg diet), a nitrogen-free diet group and a fast group. The true nitrogen-corrected ME (TME$_n$) and true amino acid availability were determined. In experiment 2, 324 day-old chicks were used in a 2${\times}$3 factorial design consisting of two energy contents (high and low) and three GAL levels (0, 250, and 500 mg/kg). Three feeding phases, comprising 0-21 d, 22-35 d and 36-48 d, were involved. GAL addition improved TME$_n$ and the availability of methionine and cystine (p<0.05). The apparent ME (AME) or nitrogen-corrected AME (AME$_n$) and digestibility of dry matter, organic matter, calcium, and phosphorus were improved significantly on d 21, so was crude protein and an interaction of energy and GAL on AME$_n$ (p<0.05) was found on d 35. However, daily intake and daily gain were significantly improved with GAL addition (p<0.05) during 21 d. The small intestine relative weight decreased at 250 mg/kg GAL (p<0.05) on d 35, whereas presented an interaction between GAL and energy on d 21 (p<0.05). Likewise, this treatment increased breast muscle ratio (p<0.05). On d 21, triglycerides level of broilers showed interaction between energy and enzyme levels (p<0.05). Uric acid level in 500 mg/kg GAL declined linearly (p<0.05). On d 35, quadratic effects (p<0.05) were observed in total protein, albumin, globulin and cholesterol content for enzyme supplementation. And the interactive effects of energy and GAL on serum values showed more obviously. The study implies that GAL improved energy and nutrient availability of corn-soybean meal diet in broiler. The GAL supplementation to corn-soybean meal based diet can improve performance of broilers in early stages of growth.

Studies on the Growth Characteristics of Bifidobacteria, Organic Acids and n-hexanal Contents During the Fermentation of Enzyme Treated Soy Yogurt (효소처리 분리대두단백의 요구르트 발효 중 비피더스균의 생육특성 및 유기산과 n-hexanal 함량에 관한 연구)

  • 이숙영;이정은;박미정;권영실
    • Korean journal of food and cookery science
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    • v.14 no.5
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    • pp.589-596
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    • 1998
  • This study was carried out to evaluate the quality attributes of soy yogurts prepared by different types of oligosaccharides (fructooligosaccharide, galactooligosaccharide , isomaltooligosaccharide) and Bifidobacteria (B. bifidum B. breve, B. infantis) containing enzyme treated soy protein isolate in terms of pH, titratable acidity, total number of viable cells of Bifidobacteria, ${\alpha}$-galactosidase activity, organic acids, volatile compounds. The pH values of soy yogurts fermented by B. bifidum showed the highest significantly but those fermented by B. infantis showed the lowest significantly, while the titratable acidity of soy yogurts were vice versa. The viable cells of Bifidobacteria of all soy yogurts showed more than 10$\^$9/ CFU/ml and soy yogurts fermented by B. infantis showing below pH 4.6 showed more than 10$\^$9/ CFU/ml after storage at 4$^{\circ}C$ for 7 days. The activity of ${\alpha}$-galactosidase showed the highest in the culture of B. infantis among the Bifidobacteria tested. Among the Bifidobacteria tested, the contents of lactic acid and acetic acid showed the highest in soy yogurts fermented by B. infantis but citric acid and propionic acid were the lowest. Among the Bifidobacteria tested, the contents of n-hexanal showed the highest in soy yogurts fermented by B. breve and a little amounts of acetaldehyde were present in all soy yogurts.

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Survey of the Expression Pattern and Immuno Stimulatory Effect of DNA Vaccine Using β-Galactosidase Reporter System in Olive Flounder (Paralichthys olivaceus)

  • Lee Sang-Jun;Hong Suhee;An Kyong-Jin;Kim Young-Ok
    • Fisheries and Aquatic Sciences
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    • v.7 no.2
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    • pp.70-75
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    • 2004
  • The CMV promoter driven lacZ reporter gene (pcDNA-lacZ) was constructed and used for DNA immunization study. The expression of the lacZ gene was confirmed in vitro using RTG-2 cell line before using for in vivo study in olive flounder (Paralichthys olivaceus). In the dose response study, the maximum expression of the lacZ gene was found in the group injected with 5 ${\mu} g$ of the plasmid DNA. Kinetic study showed a significantly increased expression of $\beta-galactosidase$ gene at 7 days after injection. Effects of DNA vaccine on specific and nonspecific immune responses such as antibody and NO production were studied and the significant effect was found in olive flounder injected with 10 and 15 ${\mu} g$ DNA (sub optimal dose for lacZ gene expression). Two pro inflammatory cytokine genes, $IL-l\beta$ and $TNF-\alpha$, were also found to be up regulated in the muscle injected with the plasmid, suggesting an induction of local inflammatory response.

Efficacy of Enzyme Treatment for the Quality Improvement of Soymilk (두유(豆乳)의 품질향상을 위한 효소제(酵素劑) 처리의 효과)

  • Yoo, Jeong-Seon;Lee, Su-Rae
    • Korean Journal of Food Science and Technology
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    • v.20 no.3
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    • pp.426-432
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    • 1988
  • This study was undertaken to investigate for the possible use of enzymes with ${\alpha}-galactosidase$ and protease activities to remove flatulence factors as well as to improve the yield and protein digestibility in soymilk preparation. The volume and protein yield were not increased significantly by enzyme treatment. The solids yield increased by raising treatment pH 6 to 10, the temperature $30^{\circ}C\;to\;60^{\circ}C$ Enzyme treatment brought about a remarkable increase in TCA-soluble nitrogen compounds and a decrease in the contents of flatulence factors raffinose and stachyose.

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Enzymatic Synthesis and Characterization of Galactosyl Trehalose Trisaccharides

  • Kim, Bong-Gwan;Lee, Kyung-Ju;Han, Nam-Soo;Park, Kwan-Hwa;Lee, Soo-Bok
    • Food Science and Biotechnology
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    • v.16 no.1
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    • pp.127-132
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    • 2007
  • [ ${\alpha},\;{\alpha}$ ]-Trehalose was efficiently modified by a transgalactosylation reaction of Escherichia coli ${\beta}-galactosidase$ using lactose as a donor to yield two galactosyl trehalose trisaccharides. The reaction products of trehalose by the enzyme were observed by thin layer chromatography (TLC) and high performance anion exchange chromatography (HPAEC) and were purified by BioGel P2 gel permeation chromatography and recycling preparative HPLC. Liquid chromatography-mass spectrometry (LC-MS) and ^{13}C$ nuclear magnetic resonance (NMR) analyses revealed that the structures of the main products were $6^2-{\beta}-D-galactosyl$ trehalose (1) and $4^2-{\beta}-D-galactosyl$ trehalose (2). A reaction of 30%(w/v) trehalose and 15%(w/v) lactose at pH 7.5 and $45^{\circ}C$ resulted in a total yield of approximately 27-30% based on the amount of trehalose used. The galactosyl trehalose products were not hydrolyzed by trehalose. In addition the mixture of transfer products (9:1 ratio of 1 to 2) showed higher thermal stability than glucose, lactose, and maltose, but less than trehalose, against heat treatment over $100^{\circ}C$ at pH 4 and 7. It also exhibited better thermal stability than sucrose at pH 4 alone.