• Title/Summary/Keyword: P3a

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Pilot Scale Production of Poly (3-Hydroxybutyrate-co-3-hydroxy-valerate) by Fed-batch Culture of Recombinant Escherichia coli

  • Park, Jong-il;Lee, Sang-Yup;Kyungsup Shin;Lee, Woo-Gi;Park, Si-Jae;Chang, Ho-Nam;Chang, Yong-Keun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.6
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    • pp.371-374
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    • 2002
  • Production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate)[P(3HB/V)], by fed-batch culture of recombinant Escherichia coli harboring a plasmid containing the Alcaligenes latus polyhy-droxyalkanoate (PHA) biosynthesis genes, was examined in two pilot-scale fermentors with air supply only, In a 30 L fermentor having a XLa value of 0.11 S­$^1$, the final P(3HB/V) concentration and the P(3HB/V) content obtained were 29.6 g/L and 70.1 wt%, respectively giving a productivity of 1.37 g P(3HB/V)/L-h. In a 300 L fermentor having a XLa of 0.03 S­$^1$, the P(3HB/V) concentration and the P(3HB/V) content were 20.4 g/L and 69 wt%, respectively giving a productivity of 1.06g P(3HB/V)/L-h. These results suggest that economical production of P(3HB/V) is possible by fed-batch culture of recombinant E. coli in a large-scale fermentor having low KLa value.

Effects of the Electroless Ni-P Thickness and Assembly Process on Solder Ball Joint Reliability (무전해 Ni-P 두께와 Assembly Process가 Solder Ball Joint의 신뢰성에 미치는 영향)

  • Lee, Ji-Hye;Huh, Seok-Hwan;Jung, Gi-Ho;Ham, Suk-Jin
    • Journal of Welding and Joining
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    • v.32 no.3
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    • pp.60-67
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    • 2014
  • The ability of electronic packages and assemblies to resist solder joint failure is becoming a growing concern. This paper reports on a study of high speed shear energy of Sn-4.0wt%Ag-0.5wt%Cu (SAC405) solder with different electroless Ni-P thickness, with $HNO_3$ vapor's status, and with various pre-conditions. A high speed shear testing of solder joints was conducted to find a relationship between the thickness of Ni-P deposit and the brittle fracture in electroless Ni-P deposit/SAC405 solder interconnection. A focused ion beam (FIB) was used to polish the cross sections to reveal details of the microstructure of the fractured pad surface with and without $HNO_3$ vapor treatment. A scanning electron microscopy (SEM) and an energy dispersive x-ray analysis (EDS) confirmed that there were three intermetallic compound (IMC) layers at the SAC405 solder joint interface: $(Ni,Cu)_3Sn_4$ layer, $(Ni,Cu)_2SnP$ layer, and $(Ni,Sn)_3P$ layer. The high speed shear energy of SAC405 solder joint with $3{\mu}m$ Ni-P deposit was found to be lower in pre-condition level#2, compared to that of $6{\mu}m$ Ni-P deposit. Results of focused ion beam and energy dispersive x-ray analysis of the fractured pad surfaces support the suggestion that the brittle fracture of $3{\mu}m$ Ni-P deposit is the result of Ni corrosion in the pre-condition level#2 and the $HNO_3$ vapor treatment.

Studies of Pharmacological Activity on New Oral Cephalosporins (새로운 경구용 세팔로스포린의 약효평가)

  • La, Sung-Bum;Kim, Wan-Joo;Jee, Ung-Kil
    • YAKHAK HOEJI
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    • v.38 no.2
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    • pp.140-148
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    • 1994
  • ${\beta}-Lactamase$ stability, chemotherapeutic activity, and pharmacokinetics of 7-[(Z)-2-(2-aminothiazole-4-yl)-2-methoxyiminoacetamido]-3-[4-(2-pyridyl)piperazinyl]thiocarbonylthiomethyl-3-cephem-4-carboxylic acid(CEN1), 7-[(Z)-2-(2-aminothiazole-4-yl)-2-methoxyiminoacetamido]-3-[4-(2-pyrimidyl)piperazinyl]thiocarbonylthiomethyl-3-cephem-4-carboxylic acid(CEN2), pivaloyloxymethyl-7-[(Z)-2-(2-aminothizaole-4-yl)-2-methoxyiminoacetamido]-3-[4-(2-pyridyl)piperazinyl]thiocarbonyl-thiomethyl-3-cephem-4-carboxylate(CEN1P), and pivaloyloxymethyl-7-{(Z)--2-(2-aminothizaole-4-yl)-2-methoxyiminoacetamido]-3-[4-(2-pyridyl)piperazinyl]thiocarbonyl-thiomethyl-3-cephem-4-carboxylate(CEN2P) were examined. CEN1, CEN2, CEN1P, and CEN2P were very stable to the ${\beta}-lactamase$ obtained from three strains(Enterobacter cloacae P99, Escherichia coli TEM, and Citrobacter freundii). Chemotherapeutic activities$(ED_{50})$ of CEN2 and CEN2P against experimental systemic infections due to Streptococcus pyogenes 77A and Escherichia coli 078 were superior to those of CEN1 and CEN1P, respectively. The $ED_{50}$ values of CEN1, CEN2 were 5.82 mg/kg, 0.89 mg/kg(s.c., S. pyogenes 77A) while those of CEN1P, CEN2P were 14.56mg/kg, 6.40mg/kg(p.o., S. pyogenes 77A), respectively. The pharmacokinetics of CEN1, CEN2, CEN1P, and CEN2P were investigated in mice and rats. In mice, peak blood levels of $1.25\;{\mu}g/ml$ were recorded within 20 min after oral administration of a single dose equivalent to 40 mg/kg CEN1P. Cmax of CEN1P was much higher than that of CEN1 in mice and rats. Oral absorption of CEN2P was much higher than that of CEN2.

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Phosphorylation of 44-kilodalton Proteins in Peripheral T-lymphocyte of Rat (흰쥐 말초 혈액 림프구의 분자량 44 kD 단백의 인산화)

  • Ahn, Young-Soo;Jou, Il-O;Oh, Do-Yeun;Lim, Seung-Wook;Park, Kyung-Sun
    • The Korean Journal of Pharmacology
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    • v.27 no.2
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    • pp.135-144
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    • 1991
  • Using T-lymphocytes obtained from rat peripheral blood, we found that the 44kD/pI6.8 protein was the major phosphoprotein of T-lymphocytes under basal condition, and that the 44kD/pI6.3 protein was a new phosphoprotein appeared in T-lymphocytes stimulated with ${\beta}-agonist$. The phosphorylation of the 44kD/pI6.3 protein was also induced by forskolin but inhibited by H-8 pretreatment. To clarify the character of the 44kD/pI6.3 protein, we used Con-A and kinase inhibitors, H-7 and W-7. Con-A stimulation induced phosphorylation of 44kD/pI 6.3 protein but that was inhibited by W-7 pretreatment. The phosphorytation of 44kD/pI6.3 protein was not induced by the PKC activator, PMA. Instead, the phosphorylation of 44kD/pI6.8 protein was reduced by H-7, a PKC inhibitor. From the above results,it can be concluded that the 44kD/pI6.3 protein can be a common substrate for A-kinase and CaM kinase. The two dimensional tryptic peptide mapping revealed that the 44kD/pI6.8 and 44kD/pI6.3 proteins are different.

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Studies on Benzo(a) pyrene Concentrations in Atmospheric Particulate Matters (大氣浮游粒子狀物質中 Benzo(a) pyrene 濃度에 關한 硏究)

  • 손동헌;허문영;남궁용
    • Journal of Korean Society for Atmospheric Environment
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    • v.3 no.2
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    • pp.11-17
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    • 1987
  • Atmospheric particulate matter (A.P.M.) was collected on quartz fiber filters from March 1985 to February 1986 at Chung-Ang University according to particle size using Andersen high-volume air smapler, and benzo (a) pyrene concentration in these particulates were analyzed by high performance liquid chromatography. The annual arithmetic mean concentration of A.P.M. was 115.50$\mug/m^3$. The annual arithmetic mean concentrations of coarse particles and fine particles in A.P.M. were 52.54$\mum/m^3$ and 62.96$\mum/m^3$ respectively. THe annual arithmetic mean concentration of benzo(a)pyrene in A.P.M. was 1.44$ng/m^3$. THe annual arithmetic mean concentrations of benzo(a)pyrene in coarse particles and fine particles were 0.05 $ng/m^3$ and 1.39 $ng/m^3$ respectively. Thus, the concentration of benzo(a)pyrene showed maldistribution of 96.53% in fine particle. A.P.M. showed wide fluctuation according to the season. The concentration of A.P.M. was lowest in summer and high in spring and winter. Coarse and fine particle concentrations in A.P.M. were highest in spring and winter, respectively. The concentrations of benzo(a)pyrene was highest in winter and lowest in summer. The concentrations of benzo(a)pyrene in fine and coarse particles were highest in winter and spring, respectively.

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LncRNA MALAT1 Depressed Chemo-Sensitivity of NSCLC Cells through Directly Functioning on miR-197-3p/p120 Catenin Axis

  • Yang, Tian;Li, Hong;Chen, Tianjun;Ren, Hui;Shi, Puyu;Chen, Mingwei
    • Molecules and Cells
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    • v.42 no.3
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    • pp.270-283
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    • 2019
  • This study was aimed to explore if lncRNA MALAT1 would modify chemo-resistance of non-small cell lung cancer (NSCLC) cells by regulating miR-197-3p and p120 catenin (p120-ctn). Within this investigation, we totally recruited 326 lung cancer patients, and purchased 4 NSCLC cell lines of A549, H1299, SPC-A-1 and H460. Moreover, cisplatin, adriamycin, gefitinib and paclitaxel were arranged as chemotherapies, and half maximal inhibitory concentration (IC50) values were calculated to evaluate the chemo-resistance of the cells. Furthermore, mice models of NSCLC were also established to assess the impacts of MALAT1, miR-197-3p and p120-ctn on tumor growth. Our results indicated that MALAT1 and miR-197-3p were both over-expressed within NSCLC tissues and cells, when compared with normal tissues and cells (P < 0.05). The A549, H460, SPC-A-1 and SPC-A-1 displayed maximum resistances to cisplatin ($IC50=15.70{\mu}g/ml$), adriamycin ($IC50=5.58{\mu}g/ml$), gefitinib ($96.82{\mu}mol/L$) and paclitaxel (141.97 nmol/L). Over-expression of MALAT1 and miR-197-3p, or under-expression of p120-ctn were associated with promoted viability and growth of the cancer cells (P < 0.05), and they could significantly strengthen the chemo-resistance of cancer cells (P < 0.05). MALAT1 Wt or p120-ctn Wt co-transfected with miR-197-3p mimic was observed with significantly reduced luciferase activity within NSCLC cells (P < 0.05). Finally, the NSCLC mice models were observed with larger tumor size and weight under circumstances of over-expressed MALAT1 and miR-197-3p, or under-expressed p120-ctn (P < 0.05). In conclusion, MALAT1 could alter chemo-resistance of NSCLC cells by targeting miR-197-3p and regulating p120-ctn expression, which might assist in improvement of chemo-therapies for NSCLC.

Increase of a Fibrinolytic Enzyme Production through Promoter Replacement of aprE3-5 from Bacillus subtilis CH3-5

  • Yao, Zhuang;Meng, Yu;Le, Huong Giang;Lee, Se Jin;Jeon, Hye Sung;Yoo, Ji Yeon;Kim, Jeong Hwan
    • Journal of Microbiology and Biotechnology
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    • v.31 no.6
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    • pp.833-839
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    • 2021
  • Bacillus subtilis CH3-5 isolated from cheonggukjang secretes a 28 kDa protease with a strong fibrinolytic activity. Its gene, aprE3-5, was cloned and expressed in a heterologous host (Jeong et al., 2007). In this study, the promoter of aprE3-5 was replaced with other stronger promoters (Pcry3A, P10, PSG1, PsrfA) of Bacillus spp. using PCR. The constructed chimeric genes were cloned into pHY300PLK vector, and then introduced into B. subtilis WB600. The P10 promoter conferred the highest fibrinolytic activity, i.e., 1.7-fold higher than that conferred by the original promoter. Overproduction of the 28 kDa protease was confirmed using SDS-PAGE and fibrin zymography. RT-qPCR analysis showed that aprE3-5 expression was 2.0-fold higher with the P10 promoter than with the original promoter. Change of the initiation codon from GTG to ATG further increased the fibrinolytic activity. The highest aprE3-5 expression was observed when two copies of the P10 promoter were placed in tandem upstream of the ATG initiation codon. The construct with P10 promoter and ATG and the construct with two copies of P10 promoter in tandem and ATG exhibited 117% and 148% higher fibrinolytic activity, respectively, than that exhibited by the construct containing P10 promoter and GTG. These results confirmed that significant overproduction of a fibrinolytic enzyme can be achieved by suitable promoter modification, and this approach may have applications in the industrial production of AprE3-5 and related fibrinolytic enzymes.

Characterization of the bacteriophage P4 sid+ derivative overcoming P2sir-associated helper inefficiency through DNA conformational adaptation (DNA 형태 적응을 거쳐 P2sir-관련 도움파지 비효율성을 극복하는 박테리오파지 P4 sid+ 유도체 정성 연구)

  • Kim, Kyoung-Jin
    • Korean Journal of Microbiology
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    • v.52 no.1
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    • pp.120-124
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    • 2016
  • A certain size of DNA (28-29 kb long) to be packaged into P2-size head and the mutation in sid gene of bacteriophage P4 are the major factors to overcome "P2 sir-associated helper inefficiency". To clarify whether the presence of sid mutation is essential to overcome "P2 sir-associated helper inefficiency" or not, we tested the P4 derivative, P4 delRI::kmr, which is $sid^+$ and whose genome size supposed to be 28.5 kb long in the case of being packaged into $P2_{sir3}$-sized large head. As P4 delRI::kmr showed the low EOP with P2 sir3 lysogen, P4 delRI::kmr phage stock was prepared in P2 sir3 lysogen host to increase the EOP with P2 sir3 lysogen. Through this process, P4 delRI::kmr had been adapted for P2 sir3 lysogen. With a CsCl buoyant equilibrium density gradient experiment and gel electrophoresis of the isolated DNA, it was evident that the adaptation of P4 delRI::kmr for P2 sir3 lysogen was caused by the conformational change of DNA to be packaged into large head. The burst size determination experiments with P4 delRI::kmr phage stock adapted for P2 sir3 lysogen and normal P4 delRI::kmr phage stock showed that not the sid mutation but the size of DNA to be packaged (28-29 kb long) was essential to overcome "P2 sir-associated helper inefficiency".

Effects of $aroP^{-}$ mutation on the tryptophan excretion in escherichia coli ($aroP^{-}$변이가 E.coli에서 트립토판 방출에 미치는 영향)

  • 지연태;안병우;이세영
    • Korean Journal of Microbiology
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    • v.23 no.1
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    • pp.9-12
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    • 1985
  • As a part of the host cell development for a amplified recombinant trp operon, $aroP^-$ mutation was introduced in a E. coli host strain. $aroP^-$ mutation was induced by transposon Tn10 and transduced into the E. coli host cell by bacteriophage P1Kc. The effect of $aroP^-$ mutation on the excretion of tryptophan in E. coli $trpR^{-ts}/ColE_1 -trp^+$ cells was investigated. Mutant lacking the general aromatic transport system was resistant to ${\beta}-2-thienylalanine\;(2{\times}10^{-4}\;M)$, p-fluorophenylalanine $(2{\times}10^{-4}M)$, or 5-methyltryptophan $(2{\times}10^{-4}\;M.)[^3H]-tryptophan$ uptake of the $aroP^-$ mutant strain was reduced considerably as compared with $aroP^+$ counterpart. The rate of $[^3H]-tryptophan$ uptake of the $aroP^-$ mutant strain treated with $NaN_3(3{\times}10^{-2}\;M)$ was much less affected than that of $aroP^+$ counterpart. The $aroP^-$ transductants increased the tryptophan excretion from E. coli $trpR^{-ts}/ColE_1 -trp^+$ four times more than $aroP^+$ counterpart.

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Improved Air Stability of OTFT's with a P3HT/POSS Active Layer (P3HT/POSS 합성 활성층을 이용한 OTFT 소자의 대기안정성 향상)

  • Park, Jeong-Hwan;Han, Kyo-Yong
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.22 no.2
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    • pp.107-113
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    • 2009
  • In order to improve air stability, we proposed a new active layer of an organic TFT by synthesizing P3HT/POSS conjugated polymer. P3HT/POSS OTFTs with the various P3HT/POSS volume ratios were fabricated and characterized. With the P3HT/POSS volume ratio of 1:1, we achieved the field-effect mobilities of ${\sim}1.19{\times}10^{-3}\;cm^2/v{\cdot}sec$ in the saturation region and the current on/off ratio of ${\sim}2.51{\times}10^2$. The resulting current on-off ratio was much higher than that of the P3HT-based OTFTs and resulted from the dramatic decrease of the off-current. Since the off-current can be reduced by preventing oxygen in atmosphere from doping the P3HT/POSS active layers, this new active layer shows its ability to avoid oxygen doping in atmosphere. Therefore, the improvement of the air stability can be achieved by employing the P3HT/POSS active layers.