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Purification and Properties of $\alpha$-Glucosidase from Mococcus halophilus (Pediococcus halophilus로부터 생성한 $\alpha$-Glucosidase의 정제 및 특성)

  • 민해기;이호근;문지웅;강국희
    • Microbiology and Biotechnology Letters
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    • v.20 no.2
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    • pp.143-149
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    • 1992
  • A bacterial strain No. 2, which highly produced a-glucosidase, was isolated from Kimchi and identified to be a similar species of Pediococcus halophilus. This enzyme was purified by protamine sulfate, ammonium sulfate fractionation, ion exchange and gel filtration. The maximal a-glucosidase activity was observed at pH 6.0 and this enzyme was stable at pH 6.0~ 7.5. The optimum temperature of this enzyme activity was $37^{\circ}C$, but enzyme activity was gradually lost above $37^{\circ}C$. This enzyme was activated by 10 mM MgCh and inhibited by 10 mM mercaptoethanol. The kinetics of PNPG(p-Nitrophenyl-a-D-glucopyranoside) and maltose were Kp0.52 mM/27.5 pg protein, $V_{max}$= 0.021 mM/min 27.5 ${\mu}g$ protein and $K_m$= 0.32 mMD7.5 ${\mu}g$ protein, $V_{max}$= 0.025 mM/min 27.5 ${\mu}g$ protein, respectively. The molecular weight of $\alpha$-glucosidase was about 37, 000.

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The Study of Optimal Conditions for Synthesis and Purification of 1, 2-Octanediol Galactoside (1, 2-Octanediol Galactoside 합성을 위한 최적 조건 및 정제 연구)

  • Jung, Kyung-Hwan
    • Journal of the Korean Applied Science and Technology
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    • v.39 no.1
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    • pp.1-9
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    • 2022
  • 1, 2-Octanediol (OD) as a cosmetic additive has been used simultaneously as a preservative and humectant. To solve the skin problem by 1, 2-octanediol (OD), we have synthesized 1, 2-octanediol galactoside (OD-gal) using Escherichia coli β-galactosidase (β-gal). Meanwhile, the optimal amount of β-gal, OD concentration, pH, and temperature for OD-gal synthesis were 4.5 U/ml, 150 mM, 7.0, and 37℃, respectively. Under these conditions, 150 mM OD was converted into about 55.9 mM OD-gal during 24 hours, in which the conversion yield (mole basis) was about 37.2%. In addition, OD-gal of 67.4 mg could be purified from a 9 ml reaction mixture, in which the overall synthesis yield from OD to the purified OD-gal was about 34.1% (weight basis) and 16.2% (mole basis), respectively. We are expecting that these results will be helpful to develop a safer additive in the cosmetic industry as basic data.

A Study on Characteristics of Sn-37Pb and Sn-4.0Ag-0.5Cu Solder Joints as Various A:V Ratio (A:V Ratio 변화에 따른 Sn-37Pb, Sn-4.0Ag-0.5Cu Solder 접합부의 특성 연구)

  • Han, Hyun-Joo;Lim, Seok-Jun;Moon, Jung-Tak;Lee, Jin
    • Proceedings of the International Microelectronics And Packaging Society Conference
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    • 2001.11a
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    • pp.67-73
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    • 2001
  • To investigate the relationships of solder joint characteristics with solder composition and A:V ratio (solder volume per pad area), Sn-37Pb and Sn-4.0Ag-0.5Cu solder balls with 330, 400, 450 and $457{\mu}{\textrm}{m}$ size were reflowed on same substrate. Sn-37Pb and Sn-4.0Ag-0.5Cu was reflowed at $220^{\circ}C$ and $240^{\circ}C$ respectively by IR-type soldering machine. As a result of reflowed solder- ball diameter(D) and height(H) measurement, D/H was decreased with solder ball size increment in range of 330~450 ${\mu}{\textrm}{m}$. But, D/H was increased in the solder joint for 457 ${\mu}{\textrm}{m}$ size, it was caused possibly by decrement of solder ball height increment compared with solder volume increment. As a result of shear and pull test, joint strength with A:V ratio was high. Joint strength of Sn-4.0Ag-0.5Cu was higher than Sn-37Pb. However, Sn-37Pb had more stable solder joint of small standard deviation. A thick and clean scallop type Ni-Cu-Sn intermetallic compound layer was formed in high A:V ratio and Sn-4.0Ag-0.5Cu solder joint interface.

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Thiosulfate Oxidation and Mixotrophic Growth of Methylobacterium goesingense and Methylobacterium fujisawaense

  • Anandham, R.;Indiragandhi, P.;Madhaiyan, M.;Chung, Jong-Bae;Ryu, Kyoung-Yul;Jee, Hyeong-Jin;Sa, Tong-Min
    • Journal of Microbiology and Biotechnology
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    • v.19 no.1
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    • pp.17-22
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    • 2009
  • The mixotrophic growth with methanol plus thiosulfate was examined in nutrient-limited mixotrophic condition for Methylobacterium goesingense CBMB5 and Methylobacterium fujisawaense CBMB37. Thiosulfate oxidation increased the growth and protein yield in mixotrophic medium that contained 150mM methanol and 20mM sodium thiosulfate, at 144 h. Respirometric study revealed that thiosulfate was the most preferable reduced inorganic sulfur source, followed by sulfite and sulfur. M. goesingense CBMB5 and M. fujisawaense CBMB37 oxidized thiosulfate directly to sulfate, and intermediate products of thiosulfate oxidation such as polythionates, sulfite, and sulfur were not detected in spent medium and they did not yield positive amplification for tested soxB primers. Enzymes of thiosulfate oxidation such as rhodanese and sulfite oxidase activities were detected in cell-free extracts of M. goesingense CBMB5, and M. fujisawaense CBMB37, and thiosulfate oxidase (tetrathionate synthase) activity was not observed. It indicated that both the organisms use the "non-S4 intermediate" sulfur oxidation pathway for thiosulfate oxidation. It is concluded from this study that M. goesingense CBMB5, and M. fujisawaense CBMB37 exhibited mixotrophic metabolism in medium containing methanol plus thiosulfate and that thiosulfate oxidation and the presence of a "Paracoccus sulfur oxidation" (PSO) pathway in methylotrophic bacteria are species dependant.

Suppressive Effects of Furonaphthoquinone NFD-37 on the Production of Lipopolysaccharide-Inducible Inflammatory Mediators in Macrophages RAW 264.7

  • Kim Min-Hee;Shin Hyun-Mo;Lee Yong Rok;Chung Eun Yong;Chang Yoon Sook;Min Kyung Rak;Kim Youngsoo
    • Archives of Pharmacal Research
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    • v.28 no.10
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    • pp.1170-1176
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    • 2005
  • 2-Methyl-2-(2-methylpropenyl)-2,3-dihydronaphthoquinone[2,3-b]furan-4,9-dione (N FD-37) is a synthetic furonaphthoquinone compound. In this study, we determined that NFD-37 could inhibit the lipopolysaccharide (LPS)-induced production of inflammatory mediators in macrophages RAW 264.7. This compound inhibited LPS-induced nitric oxide (NO) or prostaglandin (PG) $E_{2}$ production in dose-dependent manners, with $IC_{50}$ values of 7.2 ${\mu}M$ and 5.3 ${\mu}m$, respectively. As the positive controls, pyrrolidine dithiocarbamate (30 ${\mu}M$) exhibited a $57{\%}$ inhibition of NO production, and NS-398 ($1{\mu}M$) manifested a $48{\%}$ inhibition of $PGE_2$ production. The inhibitory effects of NFD-37 on NO and $PGE_2$ production were determined to occur in conjunction with the suppression of inducible NO synthase or cyclooxygenase-2 expression. NFD-37 also inhibited the production of LPS-inducible tumor necrosis factor-${\alpha}$, interleukin (IL)-$1{\beta}$ and IL-6, at $IC_{50}$ values of 4.8-8.9 ${\mu}M$. We also determined the anti-inflammatory efficacy of NFD-37 using carrageenin-induced paw edema in experimental mice.

Comparison of Results According to Reaction Conditions of Thyroglobulin Test (Thyroglobulin 검사의 반응조건에 따른 결과 비교 분석)

  • Joung, Seung-Hee;Lee, Young-Ji;Moon, Hyung-Ho;Yoo, So-yoen;Kim, Nyun-Ok
    • The Korean Journal of Nuclear Medicine Technology
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    • v.21 no.1
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    • pp.39-43
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    • 2017
  • Purpose Thyroglobulin (Tg) is a biologic marker of differentiated thyroid carcinoma (DTC), produced by normal thyroid tissue or thyroid cancer tissue. Therefore, the Tg values of DTC patients is the most specific indicator for judging whether recurrence occur or whether the remaining thyroid cancer is present. Thyroid cancer is currently the most common cancer in Korea, of which 90% is differentiated thyroid cancer. The number of patients with thyroid disease of this application also increased, and an accurate and prompt results are required. However, the incubation time of the Tg commonly takes about 24 hours in our hospital, and the result reporting time is delayed, and We could not satisfied with the requirements of clinical departments and patients. In order to fulfill these requirements, experiments were conducted by shortening the incubation time between company B's Kit currently in use and company C's Kit used in other hospitals. Through these experiments, we could perform the correlation with the original method and shortening method, and could find the optimum reaction time to satisfy the needs of the departments and the patients, and we will improve the competitiveness with the EIA examination. Materials and Methods In September 2016, we tested 65 patients company B's kit and company C's kit by three incubation ways. First method $37^{\circ}C$ shaking 2hr/2hr, Second method RT shaking 3hr/2hr, Third method 1hr/1hr shaking at $37^{\circ}C$. Fourth method RT shaking 3hr method which is the original method of Company C's Kit. Fifth method, the incubation time was shortened under room temperature shaking 2hr, Sixth method $37^{\circ}C$ shaking 2hr. And we performed and compared the correlation and coefficient of each methods. Results As a result of performing shortening method on company B currently in use, when comparing the Original method of company B kit, First method $37^{\circ}C$ shaking 2hr/2hr was less than Tg 1.0 ng/mL and the ratio of $R^2=0.5906$, above 1.0 ng/mL In the value, $R^2=0.9597$. Second method RT shaking 3hr/2hr was $R^2=0.7262$ less than value of 1.0 ng/mL, $R^2=0.9566$ above than value of 1.0 ng/mL. Third method $37^{\circ}C$ shaking 1hr/1hr was $R^2=0.7728$ less than value of 1.0 ng/mL, $R^2=0.8904$ above than value of 1.0 ng/mL. Forth, Company C's The original method, RT shaking 3hr was $R^2=0.7542$ less than value of 1.0 ng/mL, and $R^2=0.9711$ above than value of 1.0 ng/mL. Fifth method RT shaking 2hr was $R^2=0.5477$ less than value of 1.0 ng/mL, $R^2=0.9231$ above than value of 1.0 ng/mL. Sixth method $37^{\circ}C$ shaking 2hr showed $R^2=0.2848$ less than value of 1.0 ng/mL, $R^2=0.9028$ above than value of 1.0 ng/mL. Conclusion Samples with both values of 1.0 ng/mL or higher in both of the six methods showed relatively high correlation, but the correlation was relatively low less than value of 1.0 ng/mL. Especially, the $37^{\circ}C$ shaking 2hr method of company C showed a sharp fluctuation from the low concentration value of 1.0 ng/mL or less. Therefore, we are planning to continuously test the time, equipment, incubation temperature and so on for the room temperature shaking 2hr method and $37^{\circ}C$ shaking 1hr/1hr of company C which showed a relatively high correlation. After that, we can search for an appropriate shortening method through additional experiments such as recovery test, dilution test, sensitivity test, and provide more accurate and prompt results to the department of medical treatment, It is competitive with EIA test.

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