• 제목/요약/키워드: specific yield

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Purification and Characterization of a Thermostable Xylanase from Paenibacillus sp. NF1 and its Application in Xylooligosaccharides Production

  • Zheng, Hong-Chen;Sun, Ming-Zhe;Meng, Ling-Cai;Pei, Hai-Sheng;Zhang, Xiu-Qing;Yan, Zheng;Zeng, Wen-Hui;Zhang, Jing-Sheng;Hu, Jin-Rong;Lu, Fu-Ping;Sun, Jun-She
    • Journal of Microbiology and Biotechnology
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    • 제24권4호
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    • pp.489-496
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    • 2014
  • High levels of extracellular xylanase activity (211.79 IU/mg) produced by Paenibacillus sp. NF1 were detected when it was submerged-cultured. After three consecutive purification steps using Octyl-Sepharose, Sephadex G75, and Q-Sepharose columns, a thermostable xylanase (XynNF) was purified to homogeneity and showed a molecular mass of 37 kDa according to SDS-PAGE. The specific activity of the purified XynNF was up to 3,081.05 IU/mg with a 14.55-fold purification. The activity of XynNF was stimulated by $Ca^{2+}$, $Ba^{2+}$, DTT, and ${\beta}$-mercaptoethanol, but was inhibited by $Fe^{2+}$, $Zn^{2+}$, $Fe^{2+}$, $Cu^{2+}$, SDS, and EDTA. The purified XynNF displayed a greater affinity for oat spelt xylan with the maximal enzymatic activity at $60^{\circ}C$ and pH 6.0. XynNF, which was shown to be cellulose-free, with high stability at high temperature ($70^{\circ}C-80^{\circ}C$) and low pH range (pH 4.0-7.0), is potentially valuable for various industrial applications. The enzyme hydrolyzed oat spelt xylan to yield mainly xylooligosaccharides (95.8%) of 2-4 degree of polymerization (DP2-4). Moreover, the majority of the xylooligosacharides (DP2-4) products was xylobiose (61.5%). The thermostable xylanase (XynNF) thus seems potentially usefull in the production of xylooligosaccharides.

Characterization of Homocysteine ${\gamma}$-Lyase from Submerged and Solid Cultures of Aspergillus fumigatus ASH (JX006238)

  • El-Sayed, Ashraf S.;Khalaf, Salwa A.;Aziz, Hani A.
    • Journal of Microbiology and Biotechnology
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    • 제23권4호
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    • pp.499-510
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    • 2013
  • Among 25 isolates, Aspergillus fumigatus ASH (JX006238) was identified as a potent producer of homocysteine ${\gamma}$-lyase. The nutritional requirements to maximize the enzyme yield were optimized under submerged (SF) and solid-state fermentation (SSF) conditions, resulting in a 5.2- and 2.3-fold increase, respectively, after the last purification step. The enzyme exhibited a single homogenous band of 50 kDa on SDS-PAGE, along with an optimum pH of 7.8 and pH stability range of 6.5 to 7.8. It also showed a pI of 5.0, as detected by pH precipitation with no glycosyl residues. The highest enzyme activity was obtained at $37-40^{\circ}C$, with a $T_m$ value of $70.1^{\circ}C$. The enzyme showed clear catalytic and thermal stability below $40^{\circ}C$, with $T_{1/2}$ values of 18.1, 9.9, 5.9, 3.3, and 1.9 h at $30^{\circ}C$, $35^{\circ}C$, $40^{\circ}C$, $50^{\circ}C$, and $60^{\circ}C$, respectively. Additionally, the enzyme $K_r$ values were 0.002, 0.054, 0.097, 0.184, and 0.341 $S^{-1}$ at $30^{\circ}C$, $35^{\circ}C$, $40^{\circ}C$, $50^{\circ}C$, and $60^{\circ}C$, respectively. The enzyme displayed a strong affinity to homocysteine, followed by methionine and cysteine when compared with non-S amino acids, confirming its potency against homocysteinuria-related diseases, and as an anti-cardiovascular agent and a specific biosensor for homocysteinuria. The enzyme showed its maximum affinity for homocysteine ($K_m$ 2.46 mM, $K_{cat}\;1.39{\times}10^{-3}\;s^{-1}$), methionine ($K_m$ 4.1 mM, $K_{cat}\;0.97{\times}10^{-3}\;s^{-1}$), and cysteine ($K_m$ 4.9 m M, $K_{cat}\;0.77{\times}10^{-3}\;s^{-1}$). The enzyme was also strongly inhibited by hydroxylamine and DDT, confirming its pyridoxal 5'-phosphate (PLP) identity, yet not inhibited by EDTA. In vivo, using Swiss Albino mice, the enzyme showed no detectable negative effects on platelet aggregation, the RBC number, aspartate aminotransferase, alanine aminotransferase, or creatinine titer when compared with negative controls.

Comparative Properties of Red Ginseng Prepared with Different Cultivation Years of Fresh Ginseng Produced in Punggi Region (풍기지역 연근별 수삼의 홍삼가공 특성 비교)

  • Kwon, Joong-Ho;Kim, Kyo-Youn;Kwon, Young-Ju;Kim, Mi-Yeung;Yoon, Sung-Ran;Chung, Hun-Sik;Lee, Ki-Teak;Cho, Soon-Heang
    • Journal of the Korean Society of Food Science and Nutrition
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    • 제37권1호
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    • pp.72-75
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    • 2008
  • Quality properties of red ginseng prepared with different cultivation years of fresh ginseng produced in Punggi region were investigated. Fresh ginseng cultivated for 4, 5, or 6 years was steamed for 3.5 hr and dried for 24 hr at $60{\sim}65^{\circ}C$ and subsequently for $3{\sim}4$ days at $40^{\circ}C$ under commercial conditions. Compared to 6 years-old roots, the five years-old roots showed similar or some lower quality properties in terms of color, appearance, diameter, and inside quality, but higher ones in length and yield of prepared red ginseng. In particular, the levels of ginsenoside $Rg_3$ and $Rh_2$, which are known as specific components in red ginseng, were the highest in 5 years-old roots. The result shows that fresh ginseng of 5 years-old roots produced in Punggi region can be utilized as a raw material for the manufacture of high-quality red ginseng.

Study on the Quality Characteristics and Retarding Retrogradation of Pound Cakes containing Teff (Eragrostis tef) Flour (테프(Eragrostis tef) 분말을 첨가한 파운드케이크의 품질 특성 및 노화 억제 분석)

  • Joung, Ki Youeng;Song, Ka-Young;O, Hyeonbin;Zhang, Yangyang;Shin, So Yeon;Kim, Young-Soon
    • Journal of the East Asian Society of Dietary Life
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    • 제27권1호
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    • pp.41-49
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    • 2017
  • This study investigated the effects of teff flour on the properties of pound cakes. Five types of pound cakes were made by addition of 0% (Control), 5% (TF5), 10% (TF10), 15% (TF15), and 20% (TF20) of teff flour based on wheat flour. Specific gravity of pound cake batter showed no significant differences among samples as 0.54 (p<0.05). Baking loss was the highest in the control at 5.92% and the lowest in TF20 at 5.27%. Batter yield was the highest in TF20 at 94.73% and the lowest in control at 94.08%. Moisture content was the highest in TF20 at 23.68% and the lowest in control at 21.32%. pH showed no significant differences among samples at 7.61~7.65 (p<0.05). a-values of crumb significantly increased while L-values and b-values significantly decreased with added teff flour amounts. (p<0.05). ${\Delta}E$ values significantly increased with teff flour at 34.32~49.24 (p<0.05). Hardness was lowest in TF20 at $180.60g/cm^2$. Springiness was the lowest in TF20 at 85.37%. Cohesiveness showed no significant differences among samples (p<0.05). Chewiness was the lowest in TF20 at 172.17 g. Hardness more rapidly increased in control at $273.13{\sim}26,123.33g/cm^2$ than that of pound cakes with teff flour. Avrami exponent (n) was the highest in the control at 3.5987 and the lowest in TF20 at 1.2144. In sensory evaluation, flavor of TF20 was higher than that of the control at 4.43. Overall acceptability had a higher score in TF20 than in the control. It is considered that addition of 20% teff flour can improve the quality characteristics and retard retrogradation of pound cake.

Diagnostic Significance of the Serologic Test Using Multiple Antigens of Mycobacterium Tuberculosis by ELISA (다양한 특이결핵항원을 이용한 결핵항체 검사(ELISA)의 진단적 유용성)

  • Kim, Dae-Yun;Choi, In-Hwan;Park, Seung-Kyu;Cho, Shang-Rae;Song, Sun-Dae
    • Tuberculosis and Respiratory Diseases
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    • 제47권6호
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    • pp.757-767
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    • 1999
  • Background: Diagnosis by smear and/or cultures of the Mycobacterium tuberculosis from body fluid or biopsy specimen is "Gold standard". However the sensitivity of the direct microscopy is relatively low and culture of mycobacteria is time consuming. Despite an explosion in the techniques of rapid identification of mycobacteria by molecular genetic means, it is laborious and expensive and then rapid, inexpensive serodiagnosis is interested in diagnosis of tuberculosis. But sensitivity and specificity of known serologic antigen is not full sufficient level and then new antigen develop and combination cocktails of new developed antigens by ELISA are needed. Method: To compare the efficacy of different mycobacterial specific antigen and to assess the applicability of the combination of several different antigens in the diagnosis of tuberculosis, five ELISA tests derived 14KDa, 16KDa, 19KDa, 23KDa, 38KDa were evaluated in 57 active pulmonary patient and 24 inactive post-therapy follow up patient and 48 normal control. Results: The optical densities of ELISA test with 14KDa, 16KDa, 19KDa, 23KDa, 38KDa were significantly higher in active tuberculosis cases than in normal control(P<0.001, P<0.001, P<0.027, P<0.001, P<0.001) and those with 16KDa, 38KDa were significant higher in active tuberculosis cases than in inactive post-therapy follow up cases(P<0.01. P<0.001) and those of 14KDa, 16KDa, 23KDa, 38KDa were significant higher in inactive post-therapy follow up cases than in normal control(P<0.008. P<0.01. P<0.006. P<0.001). The sensitivity of 14KDa, 16KDa, 19KDa, 23KDa, 38KDa in active pulmonary patient cases was 42.1%, 43.9%, 15.8%, 28.0%, 70.2%, respectively and the specificity of 14KDa, 16KDa, 19KDa, 23KDa, 38KDa in active pulmonary patient cases was 95.8%, 95.8%, 91.7%, 89.6%, 93.8%, respectively. The sensitivity and specificity of combination 38KDa with 16KDa was 87% and 93.7%. Conclusion: The sensitivity and specificity of new antigens for serodiagnosis of the tuberculosis still remains limited at around 70%, which makes its a poor diagnostic tool for disease confirmation. A combination of cocktail antigens provided by cut-off value adjustment for serodiagnosis of tuberculosis some improved diagnostic yield than single antigen serologic test.

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Solid-Phase Refolding Technology in Recombinant Proteins Recovery: Application Examples to Various Biopharmaceutical Proteins (유전자재조합 단백질 회수 공정에서의 고체상 재접힘 기술: 여러 바이오의약 단백질에의 적용 사례)

  • Kim, Min Young;Suh, Chang Woo;Kim, Chang Sung;Jo, Tae Hoon;Park, Sang Joong;Choi, Won Chan;Lee, Eun Kyu
    • Korean Chemical Engineering Research
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    • 제43권2호
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    • pp.187-201
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    • 2005
  • Bioprocessing technologies utilizing 'biorecognition' between a solid matrix and a protein is being widely experimented as a means to replacing the conventional, solution-based technology. Frequently the matrices are chromatographic resins with specific functional groups exposed outside. Since the reactions of and interactions with the proteins occur as they are attached to the solid matrix, this 'solid-phase' processing has distinct advantages over the solution-phase technology. Solid-phase refolding of inclusion body proteins uses ion exchange resins to adsorb denaturant-dissolved inclusion body. As the denaturant is slowly removed from the micromoiety around the protein, it is refolded into a native, three-dimensional structure. Once the refolding is complete, the folded protein can be eluted by a conventional elution technique such as the salt-gradient. This concept was successfully extended to 'EBA (expanded bed adsorption)-mediated refolding,' in which the denaturant-dissolved inclusion body in whole cell homogenate is adsorbed to a Streamline resin while cell debris and other impurity proteins are removed by the EBA action. The adsorbed protein follows the same refolding steps. This solid-phase refolding process shows the potential to improve the refolding yield, reduce the number of processing steps and the processing volume and time, and thus improve the overall process economics significantly. In this paper, the experimental results of the solid-phase refolding technology applied to several biopharmaceutical proteins of various types are presented.

Hydrogen Production for PEMFC Application in Plasma Reforming System (PEMFC용 플라즈마 개질 시스템의 수소 생산)

  • Yang, Yoon Cheol;Chun, Young Nam
    • Korean Chemical Engineering Research
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    • 제46권5호
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    • pp.1002-1007
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    • 2008
  • The purpose of this paper studied the optimal hydrogen production condition of plasma reforming system to operate the PEMFC. Plasma reforming reactor used with Ni catalyst reactor at the same time, So $H_2$ concentration increased. Also the WGS and PrOx reactor were designed to remove CO concentration under 10 ppm, because CO has effect on catalyst poisoning of PEMFC. The maximum $H_2$ production condition in plasma reforming system was S/C ratio 3.2, $CH_4$ flow rate 2.0 L/min, catalytic reactor temperature $700{\pm}5^{\circ}C$ and input power 900 W. At this time, the concentration of produced syngas was $H_2$ 70.2%, CO 7.5%, $CO_2$ 16.2%,$CH_4$ 1.8%. The hydrogen yield, hydrogen selectivity and $CH_4$ conversion rate were 56.8%, 38.1% and 92.2% respectively. The energy efficiency and specific energy requirement were 37.0%, 183.6 kJ/mol. In additional, The experiment of $CO_2/CH_4$ ratio proceeded. Also WGS reactor experiment was proceeding on optimum condition of plasma reactor and the exit concentration were $H_2$ 68%, CO 337 ppm, $CO_2$ 24.0%, $CH_4$ 2.2%, $C_2H_4$ 0.4%, $C_2H_6$ 4.1%. At this time, experiment result of PrOx reactor were $H_2$ 51.9%, CO 0%, $CO_2$ 17.3%.

Drivers' Understanding of Traffic Pavement Markings (교통노면표시 이해도에 관한 연구)

  • Shin, Kangwon
    • KSCE Journal of Civil and Environmental Engineering Research
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    • 제33권2호
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    • pp.711-718
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    • 2013
  • Traffic pavement markings are one of primary transportation facilities that provide drivers with various road information directly. Thus, a clear understanding of traffic pavement markings is utmost important to improve traffic safety as well as to establish a proper traffic culture. However, no past studies examined drivers' understanding of traffic pavement markings in Korea. Hence, this study investigated drivers' understanding of traffic pavement markings through an elaborated administrated survey, and analyzed the relationship between various drivers' characteristics and understanding regarding pavement marking via cross-classification table and logistic model. The analysis results show that drivers have limited understanding regarding the purpose of the markings. Specifically, the average understanding of pavement markings is 57.41%: the lowest understanding is 25.88% for yield pavement marking, and the highest understanding is 91.18% for advanced pedestrian crosswalk pavement marking. This study also revealed that the understanding of some pavement markings are somewhat influenced by user group such as drivers with suspended or revoked driver licenses, but the overall understanding of pavement markings are not significantly affected by drivers' characteristics such as gender and driving experiences at ${\alpha}$=0.05. Thus, it might be desirable for policy makers to establish pavement marking-related policies for overall drivers rather than specific drivers.

Studies on the effects of radiation from radioisotopes incorporated in plant (IV) -The effects of P-32 application on the growth of buckwheat- (작물에 흡수된 방사성 동위원소의 내부 조사 효과에 관한 연구 (IV) -교맥 생장에 미치는 P-32 시용의 잔유 효과-)

  • 김길환
    • Journal of Plant Biology
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    • 제11권1호
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    • pp.7-14
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    • 1968
  • Buckwheat seeds produced in previous year (1965) in an experimental pot culture in which nine levels of P-32 ranging from $1.4{\times}10-4 to 3.3{\times}104$\mu$c/pot(as of 27 July 1965)$, with the same specific activity, had been applied to the corresponding pots respectively, were used this year(1996) in water and soil culture as well as in germination test to investigate the feature and extent of possible residual effects of P-32 incorporated upon germination and plant growth, and the following results were obtained: 1. Under the given experimental conditions both stimulative and inhibitory effects of radiation were observed. 2. The germination rate of the seeds was lower at the higher level of P-32 aplication ranging from $3.0{\times}103 to 3.3{\times}104$\mu$c/pot$ and higher at 0.2 $\mu$c P/pot than the control. 3. Among the seeds produced at the higher level of application about 80% was failed to germinate, owing to the radiation injury. The remaining 80% was failed to germinate, owing to the radiation injury. The remaining 80% survived the damaging effect and showed vigorous growth and increased yield. The latter group of seeds thus proved themselves to be more radioresistant than the former. 4. The survived seeds produced later more straw and root on dry weight basis. The higher the level of P-32 applied, the stronger the stimulative effect showed in vegetative growth. 5. No radiation effect on linear growth of the plants was observed in the soil culture. 6. The dry weight of straw produced showed little difference at the moderate range compared with that of control in the soil culture. At high level of application, i.e. over $\mu$c P32/pot, however, the production was increased by 12-37% of control. 7. As for the dry weight of root harvested, the P-32 treatment over 24$\mu$c P32/pot produced 82-155% more than the control, whereas little difference was observed under 2.2$\mu$c P32/pot. 8. The seed production increased in general by the P-32 treatments. Particularly at the moderate level of application the rate of increase amounted to 70% of the control. 9. Those individual plants which survived damaging effects of radiation at the germinating stage shwoed remarkable stimulative effects both in vegetative and in reproductive stage of growth.

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Purification and Characterization of the D-xylulokinase from Candida sp. L-16 (Candida sp. L-16이 생산하는 D-Xylulokinase의 정제 및 특성)

  • 이종수;주길재
    • Food Science and Preservation
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    • 제9권4호
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    • pp.429-433
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    • 2002
  • The D-xylulokinase from Candida sp. L-16 was purified through a sequence of ammonium sulfate fractionation, DEAE-cellulose chromatography, Sephadex G-100 and Sephadex G-200 gel filtration. The specific activity of the purified Dxylulokinase was increased to 23.2 fold and the yield was 11.2%. The enzyme was showed to be a single protein band by SDS-PAGE. The molecular weight of the enzyme was 150,000 dalton, this enzyme was identified to be a dimer with two subunits. The optimum conditions of the enzyme were pH 8.0 and 40$\^{C}$, respectively. The enzyme was relatively stable between pH 7.0 to pH 9.0, but it was unstable over 30$\^{C}$. The enzyme showed substrate specificity on D-xylulose, D-arabinose and D-ribose, Km value and Vmax for D-xylulose were 0.042 mM and 117 units/ml, respectively. The activation energy of the enzyme was 4.75 Kcal/mol. The one was inhibited by metabolic intermediates such as 6-phosphogluconic acid, 2-keto-gluconic acid. The enzyme was activated by EDTA and thiol compounds such as cysteine-HCI, DTT and glutathione.