• Title/Summary/Keyword: tolerance to sulfate

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Biochemical Characterization of $\small{L}$-Asparaginase in NaCl-Tolerant Staphylococcus sp. OJ82 Isolated from Fermented Seafood

  • Han, Sangwon;Jung, Jaejoon;Park, Woojun
    • Journal of Microbiology and Biotechnology
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    • v.24 no.8
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    • pp.1096-1104
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    • 2014
  • $\small{L}$-Asparaginase from gram-positive bacteria has been poorly explored. We conducted recombinant overexpression and purification of $\small{L}$-asparaginase from Staphylococcus sp. OJ82 (SoAsn) isolated from Korean fermented seafood to evaluate its biotechnological potential as an antileukemic agent. SoAsn was expressed in Escherichia coli BL21 (DE3) with an estimated molecular mass of 37.5 kDa, determined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Consistent with asparaginases in gram-negative bacteria, size-exclusion chromatography determined SoAsn as a homodimer. Interestingly, the optimal temperature of SoAsn was $37^{\circ}C$ and over 90% of activity was retained between $37^{\circ}C$ and $50^{\circ}C$, and its thermal stability range was narrower than that of commercial E. coli $\small{L}$-asparaginase (EcAsn). Both SoAsn and EcAsn were active between pH 9 and 10, although their overall pH-dependent enzyme activities were slightly different. The $K_m$ value of SoAsn was 2.2 mM, which is higher than that of EcAsn. Among eight metals tested for enzyme activity, cobalt and magnesium greatly enhanced the SoAsn and EcAsn activity, respectively. Interestingly, SoAsn retained more than 60% of its activity under 2 M NaCl condition, but the activity of EcAsn was reduced to 48%. Overall, the biochemical characteristics of SoAsn were similar to those of EcAsn, but its kinetics, cofactor requirements, and NaCl tolerance differed from those of EcAsn.

Purification and Characterization of a Novel Salt-tolerant Protease Produced by Saccharomyces sp. B101 Isolated from Baker's Dough Yeast

  • Hwang, Joo-Yeon;Kim, Sang-Moo;Heo, Seok;Kim, Cheon-Jei;Lee, Chi-Ho;Lee, Si-Kyung
    • Food Science and Biotechnology
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    • v.17 no.4
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    • pp.766-771
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    • 2008
  • The proteolytic enzyme from Saccharomyces sp. B101 was purified to homogeneity by ammonium sulfate fractionation, ultrafiltration, diethyl aminoethyl (DEAE)-Sephadex A-50 ion-exchange chromatography, and Sephadex G-100 gel filtration chromatography from the culture supernatant of Saccharomyces sp. B101. The specific activity and the purification fold of the purified enzyme were 4,688.9 unit/mg and 18, respectively. The molecular weight of the purified enzyme was estimated to be 33 kDa by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimum pH and temperature for the enzyme activity were pH 8.5 and $30^{\circ}C$, respectively. The enzyme activity was relatively stable in the pH range of 6.5-8.5 at below $35^{\circ}C$. The salt-tolerance and stability for the enzyme activity were relatively stable even at NaCl concentrations of 10 and 15%. The activity of enzyme was inhibited by $Ag^{2+}$ and $Fe^{2+}$, and activated by $Mn^{2+}$. In addition, the enzyme activity was potently inhibited by ethylenediaminetetraacetic acid (EDTA) and phenylmethyl sulfonylfluoride (PMSF). Based on these findings we concluded that the purified enzyme was a serine protease. Km and Vmax values for hammastein milk casein were 1.02 mg/mL and 278.38 unit/mL, respectively.

Comparison between the Subjective Evaluation and the Objective Evaluation of the Effect of Pain Control in the Masticatory Muscle Pain

  • Kim, Dong-Keun;Ahn, Chi-Hyuk;Hwang, Mi-Jin;Lee, Yeon-Hee;Kang, Soo-Kyung;Auh, Q-Schick;Hong, Jung-Pyo;Chun, Yang-Hyun
    • Journal of Oral Medicine and Pain
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    • v.41 no.2
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    • pp.61-71
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    • 2016
  • Purpose: This study was designed to evaluate the comparison between the subjective and the objective evaluation of pain control effect in masticatory muscle pain depending on time and dose change. Methods: The patients were recruited to this study and diagnosed according to the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD). Experimental group were divided into three groups; saline injection group (n=10), morphine 1.5 mg injection group (n=10), and morphine 3.0 mg injection group (n=10). Evaluation list was the subjective pain evaluation (visual analogue scale, McGill pain questionnaire) and the objective pain evaluation (pressure pain threshold [PPT], pressure pain tolerance [PTO]). The subjective and the objective pain evaluation were performed at the times of just before injection, 10 minutes, 30 minutes, 1 hour, 24 hours, and 48 hours after injection. Then, data were statistically analyzed. Results: The results were as follows: 1) There is no statistically significant difference between the results of the subjective and the objective pain evaluation with regard to the short-term (within 1 hour) analgesic effect of morphine sulfate. 2) However, after 1 hour of injection, while the subjective pain evaluation score still decreased, the objective pain evaluation didn't show significant changes in PPT and PTO (1 hour, p<0.05; 24 hours, p<0.01; 48 hours, p<0.001). 3) In comparison to changes in the dose, the McGill pain questionnaire was the most statistically effective method among the subjective pain evaluations (1.5 mg, p<0.05; 3 mg, p<0.01). Conclusions: Therefore, it was revealed that the subjective pain evaluation was more effective to evaluate long-term pain control, and that the McGill pain questionnaire could be an effective way to evaluate pain control depending on dose changes. It requires further investigations with time and dose extension.

Effect of Masticatory Muscle Pain Control by Morphine (Morphine에 의한 저작근 통증의 조절 효과)

  • Yoo, Sang-Hoon;Kim, Min-Jae;Chang, Joo-Yeon;Kang, Soo-Kyung;Auh, Q-Schick;Hong, Jung-Pyo;Chun, Yang-Hyun
    • Journal of Oral Medicine and Pain
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    • v.37 no.3
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    • pp.169-182
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    • 2012
  • This study was designed to evaluate the pain control effect by morphine injection to masticatory muscle pain patients. Patients with masticatory muscle pain visited the Department of Oral Medicine, Kyung Hee University Dental Hospital were recruited to this study and diagnosed by RDC/TMD. Experimental group were divided into three group; saline injection group(n=10), lidocaine injection group(n=10) and morphine injection group(n=10). Evaluation list was the subjective pain evaluation(visual analogue scale, Mc Gill pain questionnaire, pain drawing) and the objective pain evaluation(pressure pain threshold, pressure pain tolerance) and evaluation time was injection before, after 10min, 30min, 60min and then it was analyzed statistically. The results were as follows : 1. The subjective pain evaluation and the objective pain evaluation were significantly different statistically in within subject effects(p<0.001). 2. The subjective pain drawing evaluation(p<0.001) were significantly different statistically in between subject effects. 3. The objective pressure pain threshold evaluation(p=0.025) were significantly different statistically in between subject effects. 4. The morphine injection group(p=0.001) were more significantly different than the saline injection group statistically in the subject pain drawing evaluation. Therefore, it was considered that the morphine injection was effective to pain control for masticatory muscle pain patients within 60 minute.

Effects of Physical and Chemical Treatments for Reduction of Staphylococcal Phages (황색포도상구균 박테리오파지의 저감화를 위한 물리화학적 처리 효과)

  • Baek, Da-Yun;Park, Jong-Hyun;Cho, Sung-Rae;Lee, Young-Duck
    • Journal of Food Hygiene and Safety
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    • v.34 no.1
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    • pp.106-114
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    • 2019
  • The effect of physical and chemical treatments to reduce staphylococcal phages was investigated. To determine impact of physical treatment on viability of phages, two staphylococcal phages (SAP84 and SAP89) were treated with multiple heat ($55^{\circ}C$ and $60^{\circ}C$) and pH (pH4, 7, 10) conditions. Viability of SAP 84 was dramatically reduced at 60C and SAP 89 was completely inactivated at 60C within 25 min. Overall, the two phages were stable under all the pH conditions tested except for the SAP 89 at pH 10. Treatments, a 10% FAS (Ferrous Ammonium Sulfate) solution and various density of ethanol and sodium hypochlorite were used to reduce the two phages. SAP 84 was unstable in 50% and 70% ethanol. However, SAP 84 and SAP 89 showed high tolerance after exposure to 100 ppm of sodium hypochlorite which is known as an effective sterilizer. As soon as the two phages were treated with 10% FAS, which is used as a virucidal agent, they were inactivated and did not form any plaque. The result of this study provides additional evidence that staphylococcal phages can be controlled by various physicochemical treatments.

Control of Stretching of Tomato (Lycopersicon esculentum Mill.) on Cylindrical Paper Pot Seedling Using High-Salinity Potassium Fertilizers (고농도 칼륨처리를 활용한 원통형 종이포트 토마토묘의 도장억제)

  • Xu, Chan;Kim, Si Hong;Kim, Dae Hoon;Kim, Jae Kyung;Heo, Jae Yun;Vu, Ngoc Thang;Choi, Ki Young;Kim, Il Seop;Jang, Dong Cheol
    • Journal of Bio-Environment Control
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    • v.29 no.4
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    • pp.354-364
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    • 2020
  • This study was conducted to examine the potential of inducing salinity stress on cylindrical paper pot tomato seedlings to inhibit overgrowth. Potassium fertilizers, sulfate of potash (K2SO4), muriate of potash (KCl), and monopotassium phosphate (KH2PO4), were prepared as two solutions of (5 and 10) dS·m-1 salinity level, respectively, to investigate the influence on tomato (Lycopersicon esculentum Mill.) seedling growth. We also investigated the adaptability and survivability of treated tomato seedlings with high-salinity potassium (10 dS·m-1 KCl) to harsh environmental conditions (water deficit, low temperature, and storage conditions). Repeated addition of high-salinity level KCl, K2SO4, or KH2PO4 markedly decreased the dry matter of shoot and root, leaf area, and net assimilate rates (NAR) but increased the stem diameter of seedlings. Among the three sources, the relative growth rate of plant height (RGRH) was most sensitive to KCl addition; increasing salinity levels of KCl solution decreased the RGRH of seedlings. The compactness, which directly reflects the stocky growth index, increased in KCl or KH2PO4 treatments. After a week's water deficit, severely wilted seedlings were observed in control seedlings (untreated with KCl), but no wilted seedlings were observed in the KCl treated seedlings, and the relative water content (RWC) of the untreated seedlings significantly decreased by 23 %, while that of the pretreated seedlings only decreased by 8 %. The increase in ion leakage of KCl treated seedlings at low temperatures was less than that of untreated seedlings. Furthermore, there was far lower damage proportion on pretreated seedlings at (9, 12, and 15)℃ storage temperatures after 20 days, compared with on unpretreated seedlings. Our results suggest that high-salinity potassium fertilizer, especially KCl, is effective in preventing tomato seedling overgrowth, while it also improves tolerance.

Effects of High-Molecular-Weight Glutenin Subunits and Agronomic Traits on Bread Wheat Quality Parameters (밀의 고분자 글루테닌 조성과 농업 형질이 품질 특성에 미치는 영향)

  • Cha, Jin-Kyung;Shin, Dongjin;Park, Hyeonjin;Kwon, Youngho;Lee, So-Myeong;Ko, Jong-Min;Lee, Jong-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.67 no.2
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    • pp.111-120
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    • 2022
  • Improving flour quality is one of the major targets of wheat breeding programs. This study determined the optimum high-molecular-weight glutenin subunits (HMW-GS) to improve flour quality, and analyzed the correlation between agronomic and quality traits in Korea. A total of 180 wheat varieties, including 55 Korean and 125 foreign cultivars, carrying various Glu-1 alleles, were evaluated for their quality and agronomic traits. Results indicated that Glu-A1b, Glu-B1b, and Glu-D1f were the most prevailing alleles for each Glu-1 locus for Korean wheat cultivars. Korean wheat cultivars recorded shorter days to heading (DTH) and longer days to maturity (DTM) compared to foreign cultivars. In addition, an interaction effect was found between Glu-A1 and Glu-B1 alleles on several quality parameters. The combination of Glu-A1c and Glu-B1i showed a higher protein content, dry gluten content, and higher sodium dodecyl sulfate (SDS) sedimentation value than other Glu-A1×Glu-B1 combinations. Cultivars carrying Glu-A1a or Glu-A1b, Glu-B1i or Glu-B1al, and Glu-D1d for each Glu-1 locus exhibited a longer mixing time and stronger mixing tolerance. The DTM positively correlated with the protein content, gluten index and SDS sedimentation value. However, a negative correlation was observed between DTH and quality traits. Owing to the above results, this study suggests that an increase in the frequency of Glu-B1i or Glu-B1al, Glu-D1d coupled with a short DTH and long DTM could significantly improve wheat quality properties.

Sex Differences in the Pain Control by the Peripheral Opioid (성별에 따른 말초 opioid의 통증조절)

  • Bae, Sung-Jae;Kim, Wan-Su;Kang, Soo-Kyung;Auh, Q-Schick;Hong, Jung-Pyo;Chun, Yang-Hyun
    • Journal of Oral Medicine and Pain
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    • v.38 no.4
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    • pp.339-356
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    • 2013
  • This study was designed to evaluate the sex differences in the pain control effect by morphine injection to masticatory muscle pain patients. Patients with masticatory muscle pain visited the Department of Oral Medicine, Kyung Hee University Dental Hospital were recruited to this study and diagnosed by RDC/TMD. Experimental group were divided into four group each from male (n=20) and female (n=20); saline injection group (n=5), lidocaine injection group (n=5), morphine 1.5 mg injection group (n=5) and morphine 3 mg injection group (n=5). Evaluation list was the subjective pain evaluation(visual analogue scale, Mc Gill pain questionnaire, pain drawing) and the objective pain evaluation(pressure pain threshold, pressure pain tolerance) and evaluation time was injection before, after 1 hour, 24 hour, 48 hour and then it was analyzed statistically. The results were as follows : 1. The male and female were significantly different statistically morphine 3 mg group in visual analogue scale evaluation. (male: p<0.05, female: p<0.05) 2. The male and female were more significantly different statistically morphine 3 mg group than morphine 1.5 mg group in McGill pain questionnaire evaluation. (male: p<0.001, female: p<0.01) 3. The male were significantly different statistically morphine 3 mg group in pain drawing evaluation and pressure pain threshold evaluation. (PD: p<0.001, PPT: p<0.05) Therefore, it was revealed that the morphine 3 mg injection for masticatory muscle pain was effective to pain control male patients and more effect than female patients in the objective pain evaluation.

Effect of Peripheral Opioid Receptor on Masticatory Muscle Pain Control (말초 opioid 수용체에 의한 저작근 통증의 조절 효과)

  • Ko, Seok-Ho;Kang, Soo-Kyung;Auh, Q-Schick;Kim, Eun-Cheol;Hong, Jung-Pyo;Chun, Yang-Hyun
    • Journal of Oral Medicine and Pain
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    • v.38 no.2
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    • pp.161-174
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    • 2013
  • This study was designed to evaluate the pain control effect by morphine injection to masticatory muscle pain patients. Patients with masticatory muscle pain visited the Department of Oral Medicine, Kyung Hee University Dental Hospital were recruited to this study and diagnosed by RDC/TMD. Experimental group were divided into four group; saline injection group (n=10), lidocaine injection group (n=10), morphine 1.5 mg injection group (n=10) and morphine 3 mg injection group (n=10). Evaluation list was the subjective pain evaluation(visual analogue scale, Mc Gill pain questionnaire, pain drawing) and the objective pain evaluation(pressure pain threshold, pressure pain tolerance) and evaluation time was injection before, after 1 hour, 24 hour, 48 hour and then it was analyzed statistically. The results were as follows : 1. The subjective pain evaluation were significantly different statistically in morphine 3 mg group after 48 hour. (VAS: p<0.01, MGQ: p<0.001, PD: p<0.05) 2. The objective pain evaluation were significantly different statistically in morphine 1.5 mg group after 1 hour. (PPT: p<0.01, PPTol: p<0.05) 3. The morphine 3 mg group were more significantly different than lidocaine group and morphine 1.5 mg group statistically in the McGill pain questionnaire evaluation. (1h: p<0.01, 24h: p<0.01, 48h: p<0.001) Therefore, it was revealed that the morphine 3 mg injection was effective to pain control for masticatory muscle pain patients within 48 hours and more effect than lidocaine injection.