• Title/Summary/Keyword: $\alpha$-D-galactosidase

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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 Dietary Carbohydrases on Growth Performance, Nutrient Digestibility and Blood Characteristics in Finishing Pigs

  • Kim, Keun Hyoung;Cho, Jin Ho;Kim, In Ho
    • Journal of Animal Science and Technology
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    • v.55 no.4
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    • pp.289-293
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    • 2013
  • The objective of this study was to assess the effects of dietary carbohydrases on growth performance, nutrient digestibility and blood characteristics in finishing pigs. A total of 90 pigs [(Landrace ${\times}$ Yorkshire) ${\times}$ Duroc] (initial BW = $56.15{\pm}1.26kg$) were used for a 35 d feeding trial. The dietary treatments included: 1) CON (control diet), 2) MIX (CON + mixture with ${\alpha}$-galactosidase and ${\beta}$-mannanase 0.05%) and 3) MAN (CON + ${\beta}$-mannanase 0.05%). There were six replications per treatment with five pigs per pen. The average daily gain (ADG) in MIX was higher than in CON (p<0.05). No significant differences were noted in the average daily feed intake (ADFI) and feed efficiency (G:F) among dietary treatments (p>0.05). Apparent total tract digestibility (ATTD) of dry matter (DM) and energy (E) in MIX increased (p<0.05) relative to CON and MAN. The ATTD of nitrogen (N) in MIX was higher (p<0.05) than in CON. No differences in red blood cells (RBC), white blood cells (WBC), lymphocytes and IgG concentrations were observed among dietary treatments (p>0.05). In conclusion, the addition of the mixture of carbohydrases (${\alpha}$-galactosidase and ${\beta}$-mannanase 0.05%) increased ADG and nutrient digestibility in finishing pigs.

Lectin Activity and Chemical Characteristics of Escherichia coli, Lactobacillus spp. and Bifidobacterium spp. from Gastrointestinal Mucosa of Growing Pigs

  • Gao, W.;Meng, Q.X.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.6
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    • pp.863-868
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    • 2004
  • Lectin activities and chemical characteristics of Escherichia coli, Lactobacillus spp. and Bifidobacterium spp. originating from the porcine cecal mucosal layer were studied based on hemagglutination assay (HA) and hemagglutination inhibition assay (HIA). Although all the bacterial strains were able to agglutinate erythrocytes of porcine or rabbit origin, much higher HA titers were consistently observed for Lactobacillus spp. than for E. coli or for Bifidobacterium spp. A remarkable reduction in HA titers occurred by the treatment of E. coli and Lactobacillus spp. with protease or trypsin and of Bifidobacterium spp. with protease, trypsin or periodate. There were no significant effects on the HA titers of the three groups of bacteria after the treatment with lipase. Hemagglutination of E. coli was strongly inhibited by D (+)-mannose and D (+)-galactose; Lactobacillus spp. by $\alpha$-L-rhamnose and methyl-$\beta$-galactopyranoside; Bifidobacterium spp. by D (+)-alactose, $\alpha$-L-rhamnose, $\alpha$-L-fucose, L (+)-arabinose, D (+)-mannose, D (-)-fructose at a relatively low concentration (1.43 to 3.75 mg/ml). These results, combined with the enhanced HA activities of the three bacterial strains by modification of rabbit erythrocytes with neuraminidase and abolished HA activity of E. coli after treatment with $\beta$-galactosidase, indicate that it might be the glycoproteinous substances surrounding the surface of the bacterial cells that are responsible for the adhesions of these microorganisms by recognizing the specific receptors on the red blood cell.

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.

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.

Glyphosate Toxicity: III. Detection of QB Protein in Thylakoid Membrane of Tomato Apical Meristem Using an Antibody Raised from Hybrid Protein of psb A and lac Z Gene (Glyphosate 독성: III. psb A와 lac Z 유전자의 Hybrid 단백질로부터 만들어진 항체를 이용한 토마토 정단분열조직의 Thylakoid막 내 QB 단백질의 검정)

  • Kim, Tae-Wan;Amrhein, Nikolaus
    • Korean Journal of Weed Science
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    • v.15 no.3
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    • pp.206-213
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    • 1995
  • Glyphosate(N-[phosphonomethyl]glycine) applied to the assimilate-exporting leaves(i.e. third old leaf) of tomato(Lycopersicon esculentum Mil var. Moneymaker). Herbicide binding protein, QB protein(D1), has been immunoblotted using the antibodies raised against the hybrid-protein expressed by a part of spinach psb A gene cloned in frame with the 3'end of lac Z gene to allow expression of the ${\beta}$-galactosidase(EC 3.21.23) in Escherichia coli. Glyphosate has an effect on a turnover of D1 within photosystem II of thylakoid membrane. The dysfunction of D1 protein within light harvesting complex(LHC-II) seems to be a pleiotropic effect of glyphosate.

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돼지 동결정액의 배양에 따른 체외수정능력과 Glycosidase Activity의 변화

  • 황인선;정희태;양부근;김정익;박춘근
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.25-25
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    • 2003
  • 본 연구는 돼지 동결-융해 정자의 배양에 의한 체외수정능력과 glycosidase activity의 관계를 검토하였으며, 또한 돼지난자의 투명대내에서 발견된 당잔기에 대한 정자의 glycosidase 특이성을 확인하기 위하여 $\alpha$-L-fucosidase, $\alpha$-D-mannosidase, $\beta$-D-galactosidase 및 N-acetyl-$\beta$-D-glucosaminidase ($\beta$-GlcNAc'ase)의 activity를 분석하였다. 그 결과 glycosidase activity는 동결정자의 융해 후 배양하지 않았을 때보다 2시간 배양했을 때 더 높게 나타났다. $\beta$-GlcNAc'ase의 activity는 정자 배양 유무에 관계없이 다른 glycosidase 처리시보다 최소한 2배 이상 높게 나타났다. 또한 첨체반응이 유기된 정자의 비율은 glycosidase ($\alpha$-D-mannosidase; P<0.05)에 의해 영향을 받았으며 정자를 배양하지 않은 경우보다는 배양된 정자에서 높게 나타났다. 그러나 배양시간에 따른 정자의 생존성에 대해 glycosidase의 종류에 따른 유의차는 인정되지 않았다. 한편 투명대내 정자의 접착과 침입에 대한 또 다른 실험에서, 서로 다른 glycosidase가 첨가된 배양액내에서 수정된 정자가 배양시간이 길어짐에 따라 정자의 침입율은 낮아졌다($\beta$-GlcNAc'ase; P<0.05). 투명대내의 정자접착 정도는 glycosidase의 첨가시에 무첨가시보다 접착정도가 더 높았으며, 가장 높은 접착율은 $\beta$-GlcNAc'ase첨가시 나타났다. 또한 모든 glycosidase 처리시 2시간 배양한 정자보다는 배양하지 않은 정자에서 투명대에 대한 접착정도가 높게 나타났으며, $\alpha$-D-mannosidase의 처리시 유의적인 차이를 보였다(P<0.05). 본 연구의 결과, $\beta$-GlcNAc'ase가 주로 돼지정자의 원형질막내에 존재하는 것으로 추측되며, 배양된 정자에 의한 투명대 접착정도와 침입율이 낮았음에도 불구하고 glycosidase activity가 증가하는 것으로 나타났다

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Changes of Glycosidase Activity of Frozen-Thawed Spermatozoa in Human

  • Lee, Chae-Sik;Lee, Sang-Chan;Lee, Ji-Eun;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.35 no.2
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    • pp.185-190
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    • 2011
  • To evaluate the effect of spermatozoa culture on glycosidase activity of frozen-thawed spermatozoa in human, the spermatozoa were treated experimentally and assayed for activities of ${\alpha}$-L-fucosidase, ${\alpha}$-D-mannosidase, ${\beta}$-D-galactosidase and N-acetyl-${\beta}$-D-glucosaminidase (${\beta}$-GlcNAc'ase). The ${\beta}$-GlcNAc'ase activity was at least two-folds higher than other glycosidases regardless of spermatozoa incubation (p<0.05). The spermatozoa motility was decreased with incubation periods, but no effects by different glycosidases on the changes of spermatozoa motility during the various periods of incubation. In all glycosidases, the spermatozoa-zona binding rates in spermatozoa without incubation were higher than in spermatozoa incubated for 2 h (p<0.05). ${\beta}$-GlcNAc'ase is present mainly in the plasma membrane of spermatozoa frozen-thawed in human. It was also shown that the glycosidase activity was increased in all glycosidases in spite of lower sperm-zona binding by spermatozoa incubation.

Effects of Ginseng Saponin on the Lysosomal Enzyme Activities in Streptozotocin-induced Diabetic Mice (인삼 Saponin이 Lysosome 효소 활성에 미치는 영향)

  • 문창규;김희수;김강석
    • Journal of Food Hygiene and Safety
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    • v.9 no.3
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    • pp.105-109
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    • 1994
  • Lysosomal enzymes might play a most important role in the pathogenesis od diabetic microangiopathy. Some glycosidases, which participate in the catabolism of glycoprotein, are significantly decreased in diabetic mice. In search of new potential lysosomal enzyme inducers, we examined the effects of crude red-ginseng saponin fraction on N-acetyl-$\beta$-D-glucosaminidase, $\beta$-D-galactosidase and $\alpha$-D-mannosidase activities in the liver and kidney of normal and streptozotocin induced diabetic mice. It was found that i.p. administration of ginseng saponin produced the induction of lysosomal enzymes in the kidney more intensively than in the liver. The obtained results suggest the possibility that ginseng saponin might prevent the diabetic microangiopathy.

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Changes of Glycosidase Activity and Fertilizing Ability in Vitro by Incubation of Frozen-Thawed Spermatozoa in the Pig (돼지 동결정액의 배양에 따른 체외수정능력과 Glycosidase Activity의 변화)

  • 황인선;정희태;양부근;김정익;박춘근
    • Korean Journal of Animal Reproduction
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    • v.27 no.2
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    • pp.153-161
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    • 2003
  • This study has evaluated effect of the spermatozoa incubation on the glycosidase activity and fertilizing ability in vitro in the pig. To identify sperm glycosidases specific for sugar residues found in the zona pellucida of pig oocytes, the spermatozoa were treated experimentally and assayed for activities of $\alpha$-L-fucosidase, $\alpha$-D-mannosidase, $\beta$-D-galactosidase and N-acetyl-$\beta$-D-glucosaminidase ($\beta$-GlcNAc'ase). The glycosidases activity were higher in spermatozoa incubated for 2h than without incubation. The $\beta$-GlcNAc'ase activity was at least two-fold higher than other glycosidase regardless of spermatozoa incubation. In the same glycosidases, the activity had a tendency to increase as time of spermatozoa incubation was prolonged, but there were no differences in spermatozoa incubated during the various periods (4~24h). The percentages of spermatozoa that reached acrosome reaction were affected by glycosidases in the medium (P<0.05, for mannosidase), and were higher in spermatozoa with that than without incubation. On the other hand, the spermatozoa motility were decreased with incubation periods, but no effects by different glycosidases on the change of sperm motility during the various periods of incubation. In other experiment, the binding and penetration of pig spermatozoa were tested with oocytes matured in vitro in the presence of various glycosidase. The penetration rates were decreased with incubation of spermatozoa when oocytes were inseminated in medium with different glycosidases. These rates were higher in spermatozoa non-incubated than with incubation for 2h (P<0.05 for GlcNAc'ase; P<0.01 for control group). The sperm-zona binding rate in control group were higherthan in medium with glycosidases. In addition, the highest binding rate were obtained in medium with GlcNAc'ase. In all glycosidases, the sperm-zona binding rate in spermatozoa without incubation were higher than incubation for 2h. The significant differences were obtained in spermatozoa treated with $\alpha$-D-mannosidase (P<0.05). These results suggest that $\beta$-GlcNAc'ase is present mainly in the plasma membrane of pig spermatozoa. It was also shown that the glycosidase activity were increased in all glycosidases in spite of low sperm-zona binding rate and penetration rates by spermatozoa incubation.