• Title/Summary/Keyword: hydrolytic activity

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Biological characteristics of Paenibacillus polymyxa GBR-1 involved in root rot of stored Korean ginseng

  • Kim, Young Soo;Kotnala, Balaraju;Kim, Young Ho;Jeon, Yongho
    • Journal of Ginseng Research
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    • v.40 no.4
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    • pp.453-461
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    • 2016
  • Background: This study aims to describe the characterization of Paenibacillus polymyxa GBR-1 (GBR-1) with respect to its positive and negative effects on plants. Methods: The morphological characteristics of GBR-1 were identified with microscopy, and subjected to Biolog analysis for identification. Bacterial population and media optimization were determined by a growth curve. The potential for GBR-1 as a growth promoting agent, to have antagonistic activity, and to have hydrolytic activity at different temperatures was assessed. The coinoculation of GBR-1 with other microorganisms and its pathogenicity on various stored plants, including ginseng, were assessed. Results: Colony morphology, endospore-bearing cells, and cell division of GBR-1 were identified by microscopy; identification was performed by utilizing the Biolog system, gas chromatography of fatty acid methyl esters (GC-FAME). GBR-1 showed the strongest antagonistic activity against fungal and bacterial pathogens. GBR-1 cell numbers were relatively higher when the cells were cultured in brain heart infusion (BHI) medium when compared with other media. Furthermore, the starch-hydrolytic activity was influenced by GBR-1 at higher temperature compared to low temperatures. GBR-1 was pathogenic to some of the storage plants. Coinoculation of GBR-1 with other pathogens causes differences in rotting on ginseng roots. A significant growth promotion was observed in tobacco seedlings treated with GBR-1 suspensions under in vitro conditions, suggesting that its volatile organic compounds (VOCs) might play a role in growth promotion. Conclusion: The results of this study indicate that GBR-1 has both positive and negative effects on ginseng root and other stored plants as a potential biocontrol agent and eliciting in vitro growth promotion.

Characteristic Changes in Red Ginseng Fusion Cheonggukjang Based on Hydrolysis Conditions (홍삼 융합청국장의 가수분해 조건에 따른 특성변화)

  • Lee, Myung-Hee;Gu, Young-Ah;Choi, Myung-Sook;Kwon, Joong-Ho;Kim, In-Sun;Jeong, Yong-Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.8
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    • pp.1031-1037
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    • 2007
  • Changes in red ginseng fusion cheonggukjang properties under various hydrolytic conditions were investigated for its possible application to different types of food products. Among the four types of protease that were analyzed, protease (KMF -G) produced the highest hydrolysis rate, calcium binding capacity, and total phenolic compound content. In addition, the highest fibrinolytic activity and ACE inhibitory activity were also exhibited at 87.10 units and 67.17%, respectively. Among a number of different protease concentrations, a 0.02% concentration of protease (KMF-G) was found to be appropriate for the purposes of the study. The best results for red ginseng cheonggukjang hydrolysis were observed at the 60 and 90 min intervals. However, there was not a significant difference between the results at the two time points. The unpleasant odor and bitter taste associated with red ginseng fusion cheonggukjang improved with hydrolytic activity exceeding 60 min. Thus, the optimal hydrolysis time was determined to be 60 min. The total ginsenoside content of red ginseng cheonggukjang was 9.197 mg/g and the hydrolysate content was 11.707 mg/g. Based on the results, it was determined that the addition of 0.02% protease (KMF -G) and treatment for 60 min are the optimal hydrolytic conditions for red ginseng cheonggukjang to improve its biochemical characteristics, including fibronolytic activity and ACE inhibitory activity.

Expression and Characterization of a New Esterase Cloned Directly from Agrobacterium tumefaciens Genome

  • PARK HYO-JUNG;KIM YOUNG-JUN;KIM HYUNG-KWOUN
    • Journal of Microbiology and Biotechnology
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    • v.16 no.1
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    • pp.145-148
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    • 2006
  • A new functional lipolytic enzyme (AT4) has recently been found from Agrobacterium tumefaciens C58 Cereon using a genome-wide approach. The enzyme has some sequence similarity to E. coli acetyl hydrolase, Emericella nidulans lipase, Moraxella sp. lipase, Acinetobacter lwoffii esterase, and Streptomyces hygroscopicus acetyl hydrolase. However, the sequence similarities are very low (less than $25\%$), suggesting that it is a new lipase/esterase enzyme. ill the present study, intact cell of the A. tumefaciens strain was shown to have lipolytic activity on a tributyrin-LB plate. The AT4 gene was then expressed at a high level in E. coli BL21 (DE3) cells and the enzyme was purified simply by Ni-NTA column chromatography. The purified enzyme showed hydrolytic activity toward p-nitrophenyl caproate, but not toward olive oil, suggesting that the AT4 enzyme was a typical esterase rather than lipase. AT4 esterase had a maximum hydrolytic activity at $45^{\circ}C$ and pH 8.0, when p-nitrophenyl caproate was used as a substrate. It was relatively stable up to $40^{\circ}C$ and at pH 5.0-9.0. Calcium ion and EDT A did not affect the activity and thermal stability of the enzyme. As for substrate specificity, AT4 enzyme could rapidly hydrolyze acetyl and butyl groups from p-nitrophenyl esters and 1-naphthyl esters. In addition, it also released acetyl residues from acetylated glucose and xylose substrates. Therefore, this new esterase enzyme might be used as a biocatalyst in acetylation and deacetylation reactions performed in the fine chemical industry.

Cellular Biomarker of Membrane Stability and Hydrolytic Enzyme Activity in the Hemocytes of Benzo(a)pyrene-exposed Pacific oyster, Crassostrea gigas

  • Jo Qtae;Choy Eun-Jung;Park Doo Won;Jee Young-Ju;Kim Sung Yeon;Kim Yoon
    • Fisheries and Aquatic Sciences
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    • v.5 no.4
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    • pp.263-270
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    • 2002
  • The Pacific oysters, Crassostrea gigas, were stressed with different concentrations of benzo(a) pyrene and depurated to determine the hemocyte lysosomal membrane stability and hydrolytic enzymatic activity as a biomarker candidate to the chemical, using NRR (neutral red retention) and API ZYM System, respectively. The membrane damage measured as NRR decrease was significant with the increase of chemical concentration and exposure time (P<0.05), providing a possible tool for biomarker. Interestingly, the control showed intrinsic stress probably due to captive life in the laboratory, and a recovering trend was also found during the depuration. The benzo(a)pyrene-exposed oysters showed increased enzyme activities in alkaline phosphatase, esterase (C4), acid phosphatase, naphthol-AS-BI-phospho­hydrolase, $\beta$-galactosidase, $\beta$-glucuronidase, and N-acetyl- $\beta$-glucosaminidase. Of them, only two enzymes, acid phosphatase and alkaline phosphatase, showed some potential available for the generation of enzymatic biomarker in the oyster. The results are suggestive of the potential availability of the cellular and enzymatic properties as a biomarker. However, considering that a robust biomarker should be insensitive to natural stress coming from normal physiological variation, but sensitive to pollutants, a concept of intrinsic stress the animal possesses should be taken into consideration. This reflects the necessity of further research on the intrinsic stress affecting the cellular and enzymatic properties of the chemical­stressed oysters prior to using the data as a biomarker.

N-Acetylglycine Side Chain is Critical for the Antimicrobial Activity of Xanthostatin

  • Kim, Si-Kwan;Ubukata, Makoto;Isono, Kiyoshi
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.998-1000
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    • 2003
  • This study was carried out to elucidate the mode of bacteriostatic property of xanthostatin (XS), a novel depsipeptide antibiotic with an N-acetylglycine side chain and selective antimicrobial activity against Xanthomonas spp. Two biotransformed XSs were isolated by the treatment of XS with the cell lysate of Xanthomonas campestris pv. citri, a solvent partition, preparative TLC, and HPLC. Structure determination of those two biotransformed XSs demonstrated deletion of the N-acetylglycine side chain. Noteworthily, they showed no antimicrobial activity against Xanthomonas spp. This result suggests that the N-acetylglycine side chain plays a critical role in the antimicrobial activity of XS, and that the bacteriostatic property of XS is due to susceptibility of the ester bond between the hexadepsipeptide nucleus and the N-acetylglycine side chain to hydrolytic enzyme(s) produced by Xanthomonas spp.

Carbohydrate Metabolism During Germination of Ginkgo (Ginkgo biloba L.) Seed (은행나무 종자의 발아에서 탄수화물 대사)

  • 김명란
    • Journal of Plant Biology
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    • v.35 no.4
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    • pp.333-338
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    • 1992
  • Ginkgo (Ginkgo biloba L.) seeds were analyzed to determine the level of soluble sugars and insoluble starch during germination. Also the activities of the hydrolytic enzymes such as amylase, invertase and phosphatase were compared. As amylase activity was sharply increased, significant decline of starch was observed in the female gametophyte and increase of soluble sugars occurred concurrently. Invertase activity was gradually increased in cotyledon and radicle, while it was very low in dry seeds. In addition, phosphatase activity was variable only in radicle, and acid phosphatase showed higher activity than alkaline phosphatase.hatase.

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Anti-pseudomonal Activity of DA-1131, A New Carbapenem Antibiotic

  • Kim, Gye-Won;Kim, Ji-Young;Park, Seong-Hak;Lim, Joong-In;Kim, Won-Bae;Junnick Yang
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.236-236
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    • 1996
  • DA-1131은 IPM이 D2 channel 특이적인 세포외막투과성을 나타내는 것과는 달리 D2 channel 이외에 azthreonam 투과경로도 이용하는 것으로 확인되었다. IPM 감수성 및 내성인 P aeruginosa 균주들에 대하여 $\beta$-lactamase의 inducible activity, hydrolytic susceptibility, affinity를 측정한 결과, Inducible activity는 DA-1131, PPM 및 MEPM이 거의 동일하였으며, 3가지 약물 모두 가수분해에 대한 안정성을 나타내었다. 그러나 $\beta$-lactamase에 대한 affinity는 IPM이 가장 컸고, MEPM, DA-1131의 순으로 감소하였다. DA-1131은 P8P2와 PBP3를 동시에 저해하며, IPM은 PBP2의 저해 후 순차적으로 PBP3를 저해하였고 이러한 시험결과는 PBPs blinding pattern과 밀접한 관계가 있는 것으로 알려진 균형태변화로도 확인되었다.

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Biochemical Studies of an Endoglucanase from Marine Rotifer, Brachionus plicatilis

  • Chun Chang Zoon;Park Heum Gi;Hur Sung Bum;Kim Young Tae
    • Journal of Aquaculture
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    • v.9 no.4
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    • pp.453-459
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    • 1996
  • Cellulase was purified from marine rotifer, Brachionus plicatilis, to homogeneity by using chromatographic methods. Purified enzyme is an endo-${\beta}$-1,4 glucanase and shows a strong hydrolytic activity against carboxymethyl (CM) -cellulose. The physicochemical parameters of enzyme activity were determined. The molecular weight of the purified protein was approximately 62 kDa as determined by SDS-polyacrylamide gel electrophoresis. The enzymatic capability to digest cellulose of Chlorella cell wall was compared with that of other well known cellulases from Thermomonospora fusca. Experiments involving Chlorella digestion indicated that CM-cellulase from marine rotifer, Brachionus plicatilis, could digest Chlorella very efficiently while cellulase purified from Thermomonospora fusca did not. From the result here, we propose that the cellulolytic system from marine rotifer is responsible for the hydrolysis of cellulosic wall of Chlorella, probing that rotifer digests Chlorella as a major live food.

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Screenin of Phospholipase D Producing Actinomycetes (방선균으로부터 Phospholipase D 생산균주의 탐색)

  • 손동화;심재용;윤석후
    • Microbiology and Biotechnology Letters
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    • v.22 no.4
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    • pp.333-339
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    • 1994
  • In order to screen microorganisms producing phospholipase D (PLD) [EC 3.1.4.4], culture broths of about 900 strains of soil bacteria were subjected to examine for the PLD activity. When the hydrolytic activity of PLD (H-activity) in the supernatant was determined, 64 strains produced PLD more than 0.3 unit/ml and all of them were actinomycetes. Among 26 culture broths tested, 6 ones had transphosphatidylation activity (T-activity) of 30~68%. When the strains except one were cultivated on 3 different media at 30$\circ$C for 3 days under aerobic condition, strain # 1090 on medium B (yeast extract 1%, peptone 1%, glucose 1.5%, glycerol 1%, CaCO$_{3}$ 0.4%, and pH 7.2) produced PLD with much higher H- and T-activity, which were 8.3 units/ml and 76.3%, respectively. Subsequently, time course of PLD production of the strain # 1090 during cultivation with aeration of 1 v/v/m and agitation of 400 rpm at 30$\circ$C for 5 days on medium B in jar fermentor was investigated. H-activty of PLD reached almost maximum (about 9 units/ml) after 32 hours and maximal T-activity was found to be about 80%.

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Ginsenoside-Rb2 and 20(S)-Ginsenoside-Rg3 from Korean Red Ginseng Prevent Rotavirus Infection in Newborn Mice

  • Yang, Hui;Oh, Kwang-Hoon;Kim, Hyun Jin;Cho, Young Ho;Yoo, Yung Choon
    • Journal of Microbiology and Biotechnology
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    • v.28 no.3
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    • pp.391-396
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    • 2018
  • It is well known that Korean red ginseng has various biological activities. However, there is little knowledge about the antiviral activity of Korean red ginseng and its ginsenosides. In this study, we addressed whether oral administration of ginsenoside-Rb2 and -Rg3 is able to protect against rotavirus (RV) infection. The protective effect of ginsenosides against RV infection was examined using an in vivo experiment model in which newborn mice (10-day-old) were inoculated perorally (p.o.) with $1.5{\times}10^6$ plaque-forming units/mouse of RV strain SA11. When various dosages of ginsenoside-Rb2 (25-250 mg/kg) were administered 3days, 2 days, or 1 day before virus challenge, treatment with this ginsenoside at the dosage of 75 mg/kg 3days before virus infection most effectively reduced RV-induced diarrhea. In addition, consecutive administration of ginsenoside-Rb2 (75 mg/kg) at 3 days, 2 days, and 1 day before virus infection was more effective than single administration on day -3. The consecutive administration of ginsenoside-Rb2 also reduced virus titers in the bowels of RV-infected mice. In an experiment to compare the protective activity between ginsenoside-Rb2 and its two hydrolytic products (20(S)- and 20(R)-ginsenoside-Rg3), 20(S)-ginsenoside-Rg3, but not 20(R)-ginsenoside-Rg3, prevented RV infection. These results suggest that ginsenoside-Rb2 and its hydrolytic product, 20(S)-ginsenoside-Rg3, are promising candidates as an antiviral agent to protect against RV infection.