• Title/Summary/Keyword: tolerance analysis

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Characterization of Novel Salt-Tolerant Esterase Isolated from the Marine Bacterium Alteromonas sp. 39-G1

  • Won, Seok-Jae;Jeong, Han Byeol;Kim, Hyung-Kwoun
    • Journal of Microbiology and Biotechnology
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    • v.30 no.2
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    • pp.216-225
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    • 2020
  • An esterase gene, estA1, was cloned from Alteromonas sp. 39-G1 isolated from the Beaufort Sea. The gene is composed of 1,140 nucleotides and codes for a 41,190 Da protein containing 379 amino acids. As a result of a BLAST search, the protein sequence of esterase EstA1 was found to be identical to Alteromonas sp. esterase (GenBank: PHS53692). As far as we know, no research on this enzyme has yet been conducted. Phylogenetic analysis showed that esterase EstA1 was a member of the bacterial lipolytic enzyme family IV (hormone sensitive lipases). Two deletion mutants (Δ20 and Δ54) of the esterase EstA1 were produced in Escherichia coli BL21 (DE3) cells with part of the N-terminal of the protein removed and His-tag attached to the C-terminal. These enzymes exhibited the highest activity toward p-nitrophenyl (pNP) acetate (C2) and had little or no activity towards pNP-esters with acyl chains longer than C6. Their optimum temperature and pH of the catalytic activity were 45℃ and pH 8.0, respectively. As the NaCl concentration increased, their enzyme activities continued to increase and the highest enzyme activities were measured in 5 M NaCl. These enzymes were found to be stable for up to 8 h in the concentration of 3-5 M NaCl. Moreover, they have been found to be stable for various metal ions, detergents and organic solvents. These salt-tolerant and chemical-resistant properties suggest that the enzyme esterase EstA1 is both academically and industrially useful.

Cold-Stress Response of Probiotic Lactobacillus plantarum K25 by iTRAQ Proteomic Analysis

  • Liu, Shaoli;Ma, Yimiao;Zheng, Yi;Zhao, Wen;Zhao, Xiao;Luo, Tianqi;Zhang, Jian;Yang, Zhennai
    • Journal of Microbiology and Biotechnology
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    • v.30 no.2
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    • pp.187-195
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    • 2020
  • To understand the molecular mechanism involved in the survivability of cold-tolerant lactic acid bacteria was of great significance in food processing, since these bacteria play a key role in a variety of low-temperature fermented foods. In this study, the cold-stress response of probiotic Lactobacillus plantarum K25 isolated from Tibetan kefir grains was analyzed by iTRAQ proteomic method. By comparing differentially expressed (DE) protein profiles of the strain incubated at 10℃ and 37℃, 506 DE proteins were identified. The DE proteins involved in carbohydrate, amino acid and fatty acid biosynthesis and metabolism were significantly down-regulated, leading to a specific energy conservation survival mode. The DE proteins related to DNA repair, transcription and translation were up-regulated, implicating change of gene expression and more protein biosynthesis needed in response to cold stress. In addition, two-component system, quorum sensing and ABC (ATP-binding cassette) transporters also participated in cell cold-adaptation process. These findings provide novel insight into the cold-resistance mechanism in L. plantarum with potential application in low temperature fermented or preserved foods.

Influence of 3-(N-methyl-N-X(Sub.)Phenylaminooxoacetyl) Group on the Herbicidal Activity of Imazapyr Derivatives (Imazapyr 유도체의 제초활성에 미치는 3-(N-methyl-N(X)-치환-Phenylaminooxoacetyl) Group의 영향)

  • Sung, N.D.;Ryu, T.S.;Chang, H.S.;Kim, D.W.
    • Applied Biological Chemistry
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    • v.37 no.6
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    • pp.516-521
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    • 1994
  • New seventeen imazapyr derivatives, 2-(4-isopropyl-4-methyl-5-oxo-2-imidazolin-2-yl)-3-(N-methyl-N-X(sub.)-phenylaminooxoacetyl)pyridine, 6 were synthesized and their pre-emergence herbicidal activity$(pI_{50})$ in vivo against Corn (Zea mays L.) and Pigweed (Amaranthus viridis L.) were studied by the pot test under paddly conditions. Quantitative structure activity relationships (QSARs) were analyzed using the physicochemical parameters of substituent(X) on the phenyl ring of 3-(N-methyl-N-X(sub.)-phenylaminooxoacetyl) group and regression analysis. The herbicidal activities were related to the steric effect of X-substituent. The effect was rationalized by paraholic function of MR and $L_1$, where the optimal values were MR=5.56 (Zea mays L.) and $L_1=3.34\;{{\AA}}$ (Amaranthus viridis L.). Among them, 2,5-difluoro substituted compound, 6i showed good herbicidal activity against Pigweed with excellent tolerance to Corn.

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Environmental Impact Assessments along with Construction of Residential and Commercial Complex (주거단지 건설이 하천에 미치는 생태영향평가)

  • An, Kwang-Guk;Han, Jeong-Ho;Lee, Jae Hoon
    • Journal of Environmental Impact Assessment
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    • v.21 no.5
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    • pp.631-648
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    • 2012
  • The integrative ecological approaches of chemical assessments, physical habitat modelling, and multi-metric biological health modelling were applied to Gwanpyeong Stream within Gap-Stream watersheds to evaluate environmental impacts on the constructions of residential and commercial complex. For the analysis, the surveys conducted from 45 sites of reference streams within the Gap-Stream watershed and 3 regular sites during 2009 - 2010. Physical habitat health, based on the habitat model of Qualitative Habitat Evaluation Index(QHEI) declined from the headwaters(good - fair condition) to the downstream(poor condition). Chemical water quality, based turbidity and electric conductivity(EC), was degraded toward to the downstream, and especially showed abrupt increases, compared to the values of control streams(CS). Also, concentrations of chlorophyll-a in the downstreams were greater compared to the control stream(CS), indicating an eutrophication. Biological health conditions, based on the Index of Biological Integrity(IBI) using fish assemblages, averaged 19.3 which is judged as a fair condition by the biological criteria of the Ministry of Environment, Korea. The comparisons of model metric values in sensitive species and riffle-benthic species on the Maximum Species Richness Line(MSRL) of 45 reference streams indicated a massive disturbances in all sampling locations. Also, tolerance guild and trophic guild analyses suggest that dominances of tolerant species and omnivores were evident, indicating a biological degradation by habitat disturbances and organic matter pollutions. There was no distinct longitudinal variations of IBI model values from the headwater to the downstream in spite of slight chemical and habitat health gradients among the sampling sites. Overall, integrative ecological health(IEH) scores, based on the chemical, physical, and biological parameters, were low compared to the 45 reference streams due to physical and chemical disturbances of massive constructions of the residential and commercial complex. This stream, thus showed a tendency of typical urban streams which are disturbed in the chemical water quality, habitat structures, and biological integrity. Effective stream management plans and restoration strategies are required in this urban stream for improving integrative stream health.

Preventing Subway Crimes : Focusing on Sexual Offenses in the Seoul Metropolitan Subway (지하철범죄 예방전략 : 서울 지하철의 성범죄를 중심으로)

  • Roh, Sung-Hoon;Kim, Hak-Kyong
    • The Journal of the Korea Contents Association
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    • v.12 no.3
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    • pp.190-205
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    • 2012
  • The purpose of the paper is to provide policy suggestions for effectively tackling subway crimes, based on the UK and USA's crime control strategies and systems of subway crimes. The paper attempts to analyze the characteristics of subway crimes in Seoul Subways, and discovers that many of the crimes on the subways are basically 'opportunistic' crimes. In particular, crime statistics of Seoul Subways clearly show that sexual-related crimes are on the sharp increase. In this context, it is of great essence that official vigilance functions should be strengthened in order to eliminate crime opportunities in Seoul Subways. Based on such an analysis - in particular, focusing on official surveillance and response capabilities of subway passengers - the paper finally presents policy recommendations as follows: 1) an increase in the number of police officers of the Subway Police Squad, 2) an increase in the number of CCTV cameras and their effective use, 3) the development of subway riders' defence abilities, 4) strong law enforcement against minor disorder offences based on zero-tolerance policing, and 5) the reinforcement of community partnerships.

A Study for Comparison of Risk Estimates According to Extrapolating Methods of Benzo(a)Pyrene in the Ambient Air (대기중 Benzo(a) pyrene의 외삽방법에 따른 위해도 추계치의 비교 연구)

  • Kim, Jong-Man;Chung, Yong
    • Journal of Korean Society for Atmospheric Environment
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    • v.8 no.1
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    • pp.29-37
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    • 1992
  • The risk of benzo(a)pyrene for cancer in the ambient air of Seoul was assessed by using the extrapolation methods. The average daily lifetime exposure of benzo(a)pyrene in the ambient air of Seoul was calculated at 6.97-24.30ng/$m^2$/day, which was based on the occurrence analysis of benzo(a)pyrene in the residential(Bull Kwang Dong) and traffic areas(Shin Chon) of Seoul. Using the dose scaling based on body surface area in comparisons of toxicity for extrapolation from animal to human and mathematical models from the high dose region, the low-dose risk was estimated. The response probabilities were estimated by the tolerance distribution models; Probit, Logit and Weibull model. They were consistent with the observed ones at experimental dose region. The unit risk estimates of these models were too low to be used. One-hit and multistage model to prove more conservative risk was selected. As a redult, the lifetime unit risk of benzo(a)pyrene for cancer and virtually safe dose were calculated; One-hit model provided the risk 2.8 $\times 10^{-7}$ and 3.4ng/$m^3$, respectively and multistage model provided 5.2 $\times 10^{-7}$ and 1.9ng/$m^3$ as the more conservatives. The lifetime excess risk estimates of benzo(a)pyrene for cancer were calculated at 0.37-1.30 persons/million persons by one-hit model and 0.69-2.41 persons/million persons by multistage model, which was considered in without virtual risk.

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Isolation and Biochemical Characterization of Bacillus pumilus Lipases from the Antarctic

  • Arifin, Arild Ranlym;Kim, Soon-Ja;Yim, Joung Han;Suwanto, Antonius;Kim, Hyung Kwoun
    • Journal of Microbiology and Biotechnology
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    • v.23 no.5
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    • pp.661-667
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    • 2013
  • Lipase-producing bacterial strains were isolated from Antarctic soil samples using the tricaprylin agar plate method. Seven strains with relatively strong lipase activities were selected. All of them turned out to be Bacillus pumilus strains by the 16S rRNA gene sequence analysis. Their corresponding lipase genes were cloned, sequenced, and compared. Finally, three different Bacillus pumilus lipases (BPL1, BPL2, and BPL3) were chosen. Their amino acid sequence identities were in the range of 92-98% with the previous Bacillus pumilus lipases. Their optimum temperatures and pHs were measured to be $40^{\circ}C$ and pH 9. Lipase BPL1 and lipase BPL2 were stable up to $30^{\circ}C$, whereas lipase BPL3 was stable up to $20^{\circ}C$. Lipase BPL2 was stable within a pH range of 6-10, whereas lipase BPL1 and lipase BPL3 were stable within a pH range of 5-11, showing strong alkaline tolerance. All these lipases exhibited high hydrolytic activity toward p-nitrophenyl caprylate ($C_8$). In addition, lipase BPL1 showed high hydrolytic activity toward tributyrin, whereas lipase BPL2 and lipase BPL3 hydrolyzed tricaprylin and castor oil preferentially. These results demonstrated that the three Antarctic Bacillus lipases were alkaliphilic and had a substrate preference toward short- and medium-chain triglycerides. These Antarctic Bacillus lipases might be used in detergent and food industries.

Improvement of Enzymatic Stability and Catalytic Efficiency of Recombinant Fusariumoxysporum Trypsin with Different N-Terminal Residues Produced by Pichiapastoris

  • Yang, Ning;Ling, Zhenmin;Peng, Liang;Liu, Yanlai;Liu, Pu;Zhang, Kai;Aman, Aman;Shi, Juanjuan;Li, Xiangkai
    • Journal of Microbiology and Biotechnology
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    • v.28 no.9
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    • pp.1482-1492
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    • 2018
  • Fusarium oxysporum trypsin (FOT) is a fungal serine protease similar to mammal trypsin. The FOT could be successfully expressed in Pichiapastoris by engineering the natural propeptide APQEIPN. In this study, we constructed two recombinant enzymes with engineered amino acid sequences added to the N-terminus of FOT and expressed in P. pastoris. The N-terminal residues had various effects on the structural and functional properties of trypsin. The FOT, and the recombinants TE (with peptide YVEF) and TS (with peptide YV) displayed the same optimum temperature ($40^{\circ}C$) and pH (8.0). However, the combinants TE and TS showed significantly increased thermal stability at $40^{\circ}C$ and $50^{\circ}C$. Moreover, the combinants TE and TS also showed enhanced tolerance of alkaline pH conditions. Compared with those of wild-type FOT, the intramolecular hydrogen bonds and the cation ${\pi}$-interactions of the recombinants TE and TS were significantly increased. The recombinants TE and TS also had significantly increased catalytic efficiencies (referring to the specificity constant, $k_{cat}/K_m$), 1.75-fold and 1.23-fold than wild-type FOT. In silico modeling analysis uncovered that the introduction of the peptides YVEF and YV resulted in shorter distances between the substrate binding pocket (D174, G198, and G208) and catalytic triad (His42, Asp102, and Ser180), which would improve the electron transfer rate and catalytic efficiency. In addition, N-terminal residues modification described here may be a useful approach for improving the catalytic efficiencies and characteristics of other target enzymes.

Effects of warmed carrier fluid on nefopam injection-induced pain

  • Cho, Hyung Rae;Kim, Seon Hwan;Kim, Jin A;Min, Jin Hye;Lee, Yong Kyung
    • The Korean Journal of Pain
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    • v.31 no.2
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    • pp.102-108
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    • 2018
  • Background: Nefopam is a non-opioid, non-steroidal analgesic drug with fewer adverse effects than narcotic analgesics and nonsteroidal anti-inflammatory drugs, and is widely used for postoperative pain control. Because nefopam sometimes causes side effects such as nausea, vomiting, somnolence, hyperhidrosis and injection-related pain, manufacturers are advised to infuse it slowly, over a duration of 15 minutes. Nevertheless, pain at the injection site is very common. Therefore, we investigated the effect of warmed carrier fluid on nefopam injection-induced pain. Methods: A total of 48 patients were randomly selected and allocated to either a control or a warming group. Warming was performed by diluting 40 mg of nefopam in 100 ml of normal saline heated to $31-32^{\circ}C$ using two fluid warmers. The control group was administered 40 mg of nefopam dissolved in 100 ml of normal saline stored at room temperature ($21-22^{\circ}C$) through the fluid warmers, but the fluid warmers were not activated. Results: The pain intensity was lower in the warming group than in the control group (P < 0.001). The pain severity and tolerance measurements also showed statistically significant differences between groups (P < 0.001). In the analysis of vital signs before and after the injection, the mean blood pressure after the injection differed significantly between the groups (P = 0.005), but the heart rate did not. The incidence of hypertension also showed a significant difference between groups (P = 0.017). Conclusions: Use of warmed carrier fluid for nefopam injection decreased injection-induced pain compared to mildly cool carrier fluid.

Herbicidal Activities of Phenylvinylsulfone Derivatives (Phenylvinylsulfone 유도체의 제초활성)

  • Yu, Seong-Jae;Jeon, Dong-Ju;Kim, Dae-Whang;Sung, Nack-Do
    • Applied Biological Chemistry
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    • v.38 no.1
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    • pp.90-94
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    • 1995
  • Post emergence herbicidal activities$(pI_{50})$ of X-substituted phenylvinylsulfone derivatives(S) in-vivo against rice(Oryza sativa L.), Barnyard grass(Echinochloa crus-galli) and Pickerelweed(Monochoria vaginalis Presl) were measured by the pot test under paddy conditions. The (S) showed herbicidal symptom rapidly with lower activity(average $pI_{50}=2.0$) as proherbicide, which was excellent tolerance to rice. The structure activity relationships(SAR) were analyzed using such a physicochemical parameters as hydrophobic$({\pi})$ and molecular orbital(MO) quantity by the multiple regression technique, and discussed with quantum pharmacology. The herbicidal activities were related to the hydrophobic$({\pi})$ effect of X-substituent and orbital(HOMO & LUMO) energy. In case of Pickerelweed, the effect was rationalized by parabolic function of ${\pi}$ constant, where the optimal value of ${\pi}$ was 1.10. An increase in hydrophobicity and negative orbital energy by the electron attracting X-substituent may contribute to the herbicidal activity. Based on results proposed from SAR analysis, the mode of herbicidal action could be assumed.

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