• Title/Summary/Keyword: isopropyl

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Production of Aminolevulinic Acid by Recombinant Escherichia coli Co-expressing hemA and otsBA Using Crude Glycerol as Carbon Source (폐글리세롤을 탄소원으로 hemA와 otsBA를 공동 발현하는 재조합대장균 배양을 통한 아미노레블린산 생산)

  • Yan, Jingmei;Pham, Diep Ngoc;Kang, Dae-Kyung;Kim, Sung Bae;Kim, Chang-Joon
    • Microbiology and Biotechnology Letters
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    • v.44 no.3
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    • pp.341-348
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    • 2016
  • This study aimed to develop a microbial process for producing aminolevulinic acid (ALA) using crude glycerol. In the culture of ALA-producing cells (Escherichia coli/pH-hemA) in a medium containing crude glycerol, the cell density and production were 1.8-fold and 1.2-fold lower than those obtained from pure glycerol, respectively. However, the cell growth and production were improved by supplementing the medium with trehalose (30 or 100 g/l). Engineered cells (E. coli/pH-hemA/pS-otsBA) were constructed to express otsBA and their culture performance was compared with that of control cells (E. coli/pH-hemA/ pSTV28). The effects of isopropyl β-D-1-thiogalactopyranoside (IPTG) concentration and the time of induction were examined to improve the cell growth and ALA production in engineered cells cultured using crude glycerol. When 0.6 mM of IPTG was added at the beginning of the exponential growth phase, the ALA produced by cells was 2,121 mg/l, which was comparable to that from pure glycerol. The results demonstrate that otsBA expression endowed cells with the capacity to tolerate the toxicity of crude glycerol for direct use.

Synthesis of Trifluoromethylated Dihydro-1,4-oxathiin Carboxanilides and Their Fungicidal Activity (삼불화메틸기가 포함된 디히드로-1,4-옥사티인 카르복스아닐리드 유도체의 합성과 살균 활성)

  • Nam, Kee-Dal;Kim, Jin-Cheol;Cho, Kwang-Yun;Hahn, Hoh-Gyu
    • Applied Biological Chemistry
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    • v.44 no.3
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    • pp.191-196
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    • 2001
  • ${\alpha},{\beta}$-Unsaturated carboxanilides 5 with trifluromethylated dihydro-1,4-oxathiins were synthesized for the development of new agrochemical fungicide. Chlorination of trifluoromethylated ${\beta}-keto$ ester 6 followed by the reaction with 1,2-mercaptoethanol gave intermediate 1,4-oxathiane 11. Without purification of 11, substitution of hydroxy group by chlorine, followed by dehydrochlorination of 10 in the presence of triethylamine afforded trifluoromethylated dihydro-1,4-oxathiin ethyl ester 9. Acylation of the hydroxy group of the carboxylic acid 12 followed by treatment of various amines gave the corresponding trifluoromethylated dihydro-1,4-oxathiin carboxamides 5. Antifungal screening (in vivo) of the synthesized compounds against typical plant diseases, which include rice blast, rice sheath blight, cucumber gray mold, tomato late blight, wheat leaf rust, and barley powdery mildew, was carried out. Where meta position of the phenyl group was substituted with isopropoxy or isopropyl group, excellent antifungal activities against rice sheath blight and wheat leaf rust were detected.

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PROCESSING OF LIQUEFIED SARDINE PROTEIN CONCENTRATE BY ENZYMIC METHOD AND ITS UTILIZATION (산소를 이용한 정어리 액화단백질 농축물의 제조 및 이용에 관한 연구)

  • KIM Chang-Yang;HAN Bong-Ho;LEE Keun-Tai;CHO Duck-Jae;KIM Se-Kweun;KIM Soo-Hyun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.12 no.3
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    • pp.143-153
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    • 1979
  • A study on tile processing of liquefied fish protein with a long self life and good solubility has been carried out for the effective utilization of sardine. The whole sadine was chopped, homogenized with same amount of water and then hydrolyzed by the addition of commercial proteolytic enzyme. The hydrolysate was centrifuged and the supernatant was decolorized with active carbon, desodorized by azeotropic distillation with toluene, xylene and cyclohexane. The liquefied sardine protein was then concentrated by rotary vacuum evaporator with the addition of starch. The use of $0.2\%$ commercial proteolytic enzyme to the weight of the whole sardine showed the optimum hydrolysis ratio at $55^{\circ}C$ for 4 hours. The liquefied sardine protein could be decolorized and also desodorized by the treatment with $15\%$ active carl]on at room temperature for 30 minuted. In the view point of lipid concentration and the solubility of the product, the liquefied sardine protein prepared by enzymic hydrolysis from the sardine protein concentrate was better than that prepared by enzymic hydrolysis from the whole sardine and sardine protein concentrate.

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Expression and Characterization of Calcium- and Zinc-Tolerant Xylose Isomerase from Anoxybacillus kamchatkensis G10

  • Park, Yeong-Jun;Jung, Byung Kwon;Hong, Sung-Jun;Park, Gun-Seok;Ibal, Jerald Conrad;Pham, Huy Quang;Shin, Jae-Ho
    • Journal of Microbiology and Biotechnology
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    • v.28 no.4
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    • pp.606-612
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    • 2018
  • The enzyme xylose isomerase (E.C. 5.3.1.5, XI) is responsible for the conversion of an aldose to ketose, especially xylose to xylulose. Owing to the ability of XI to isomerize glucose to fructose, this enzyme is used in the food industry to prepare high-fructose corn syrup. Therefore, we studied the characteristics of XI from Anoxybacillus kamchatkensis G10, a thermophilic bacterium. First, the gene coding for XI (xylA) was inserted into the pET-21a(+) expression vector and the construct was transformed into the Escherichia coli competent cell BL21 (DE3). The expression of recombinant XI was induced in the absence of isopropyl-thio-${\beta}$-galactopyranoside and purified using Ni-NTA affinity chromatography. The optimum temperature of recombinant XI was $80^{\circ}C$ and measurement of the heat stability indicated that 55% of residual activity was maintained after 2 h incubation at $60^{\circ}C$. The optimum pH was found to be 7.5 in sodium phosphate buffer. Magnesium, manganese, and cobalt ions were found to increase the enzyme activity; manganese was the most effective. Additionally, recombinant XI was resistant to the presence of $Ca^{2+}$ and $Zn^{2+}$ ions. The kinetic properties, $K_m$ and $V_{max}$, were calculated as 81.44 mM and $2.237{\mu}mol/min/mg$, respectively. Through redundancy analysis, XI of A. kamchatkensis G10 was classified into a family containing type II XIs produced by the genera Geobacillus, Bacillus, and Thermotoga. These results suggested that the thermostable nature of XI of A. kamchatkensis G10 may be advantageous in industrial applications and food processing.

Optimization of Induction Conditions for Bacillus-derived Esterase Production by High-cell Density Fermentation of Recombinant Escherichia coli (재조합 대장균의 고농도 배양과 유도조건 최적화를 통한 Bacillus 유래 esterase의 생산)

  • Kang, Seung-Hoon;Min, Byung-Hyuk;Choi, Hong-Yeol;Kim, Dong-Il
    • Microbiology and Biotechnology Letters
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    • v.45 no.2
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    • pp.149-154
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    • 2017
  • To increase the efficiency of esterase production by Bacillus, high cell-density culture of recombinant Escherichia coli through fed batch fermentation was tested. Cells were cultured to $OD_{600}$ of 76 (35.8 g/l DCW) with dissolved oxygen level controlled to least above 30% air saturation by supplying pure oxygen. Cells were cultured to an $OD_{600}$ of 90 (42.4 g/l DCW) with glucose feeding controlled to at least 1 g/l. However, the cells reached stationary phase at the late stage of culture, despite glucose being supplied. Cells were cultured to an $OD_{600}$ of 185 (87.3 g/l DCW) by supplying additional medium with fortified yeast extract. To increase the productivity of the recombinant protein, cell growth and esterase productivity based on induction time were evaluated. Late exponential phase induction for esterase production in fed batch fermentation resulted in maximum optical density $OD_{600}$ of 190 (89 g/l DCW) and maximum esterase activity of 1745 U/l, corresponding to a 5.8-fold enhancement in esterase production, compared to the early exponential phase induction. In this study, we established fermentation methods for achieving maximum production of Bacillus-derived esterase by optimizing IPTG induction time in high-cell density culture by supplying pure oxygen and a nitrogen source.

Rejection rate and mechanisms of drugs in drinking water by nanofiltration technology

  • Ge, Sijie;Feng, Li;Zhang, Liqiu;Xu, Qiang;Yang, Yifei;Wang, Ziyuan;Kim, Ki-Hyun
    • Environmental Engineering Research
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    • v.22 no.3
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    • pp.329-338
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    • 2017
  • Nanofiltration (NF) technology is a membrane-based separation process, which has been pervasively used as the high-effective technology for drinking water treatment. In this study, a kind of composite polyamide NF thin film is selected to investigate the removal efficiencies and mechanisms of 14 trace drugs, which are commonly and frequently detected in the drinking water. The results show that the removal efficiencies of most drugs are quite high, indicating the NF is an effective technology to improve the quality of drinking water. The removal efficiencies of carbamazepine, acetaminophen, estradiol, antipyrine and isopropyl-antipyrine in ultrapure water are $78.8{\pm}0.8%$, $16.4{\pm}0.5%$, $65.4{\pm}1.8%$, $71.1{\pm}1.5%$ and $89.8{\pm}0.38%$, respectively. Their rejection rates increase with the increasing of their three-dimensional sizes, which indicates that the steric exclusion plays a significant role in removal of these five drugs. The adsorption of estradiol with the strongest hydrophobicity has been studied, which indicates that adsorption is not negligible in terms of removing this kind of hydrophobic neutral drugs by NF technology. The removal efficiencies of indomethacin, diclofenac, naproxen, ketoprofen, ibuprofen, clofibric acid, sulfamethoxazole, amoxicillin and bezafibrate in ultrapure water are $81{\pm}0.3%$, $86.3{\pm}0.5%$, $85.7{\pm}0.4%$, $93.3{\pm}0.3%$, $86.6{\pm}2.5%$, $90.6{\pm}0.4%$, $59.7{\pm}1.7%$, $80.3{\pm}1.4%$ and $80{\pm}0.5%$, respectively. For these nine drugs, their rejection rates are better than the above five drugs because they are negatively charged in ultrapure water. Meanwhile, the membrane surface presents the negative charge. Therefore, both electrostatic repulsion and steric exclusion are indispensable in removing these negatively charged drugs. This study provides helpful and scientific support of a highly effective water treatment method for removing drugs pollutants from drinking water.

Clinical Review of Toxic Alcohol Poisoning Cases in Korea (독성 알코올 중독 환자에 대한 고찰)

  • Rhee, Nu-Ga;Chung, Sung-Phil;Park, In-Cheol;Lee, Kyeong-Ryong;Kim, Hyun-Jin;Kim, Gun-Bea;Cho, Young-Soon;Kwon, In-Ho;Kim, Seung-Whan
    • Journal of The Korean Society of Clinical Toxicology
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    • v.10 no.1
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    • pp.15-21
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    • 2012
  • Purpose: Toxic alcohols are responsible for accidental and suicide motivated poisonings, resulting in death or permanent sequelae for the afflicted patients. Major therapeutic modalities in these cases include treatment with alcohol dehydrogenase inhibitors and extracorporeal elimination. There have been a number of case reports of toxic alcohol intoxication in Korea. The purpose of this study was to review the clinical characteristics of patients suffering toxic alcohol intoxication. Methods: We retrospectively reviewed the medical records of patients who presented with toxic alcohol intoxication at 8 emergency departments (ED) from Jun 2005 to Nov 2011. Patients who ingested methanol, isopropyl alcohol, ethylene glycol, and other alcohols except ethanol, were included in this study. The clinical characteristics of these patients were analyzed to include anion and osmolar gap, and estimated concentration of alcohol in the body. Results: During the study period, 21 patients were identified who had ingested toxic alcohol (methanol; 12 patients, ethylene glycol; 9 patients). At ED arrival, the mean anion gap was $18.7{\pm}6.9$ and the osmolar gap was elevated in 13 patients. Oral and IV ethanol were administrated to 11 patients in order to inhibit alcohol dehydrogenase. Extracorporeal elimination procedures such as hemodialysis were performed in 9 patients. There were no fatalities, but the one patient suffered permanent blindness. Conclusion: This study found that ethylene glycol and methanol were the substances ingested which produced toxic alcohol intoxication. The patients presented with high anion gap metabolic acidosis and were typically treated with oral ethanol and hemodialysis.

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Effect of MeOH/IPA Ratio on Coating and Fluxing of Organic Solderability Preservatives (유기 솔더 보존제의 코팅 및 플럭싱에 대한 메탄올/이소프로필알콜 비율의 영향)

  • Lee, Jae-Won;Kim, Chang Hyeon;Lee, Hyo Soo;Huh, Kang Moo;Lee, Chang Soo;Choi, Ho Suk
    • Korean Chemical Engineering Research
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    • v.46 no.2
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    • pp.402-407
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    • 2008
  • Recent popularity in mobile electronics requires higher standard on the mechanical strength of electronic packaging. Thus, the method of soldering between chip and substrate in electronic packaging process is changing from conventional method using intermetallic compound to a new method using organic solderability preservative (OSP) in order to improve the stability and the reliability of final product. Since current organic solder preservatives have several serious problems like thermo-stability during packaging process, however, it is necessary to develop new OSPs having thermo-stability. The main purpose of this study is to investigate the effect of MeOH/IPA (Isopropyl alcohol) ratio on the fluxing of a new OSP, developed in previous research, andto find out an optimum formulation of flux components for the application of the OSP in current packaging process. As a result of this study, it was revealed that higher MeOH/IPA ratio in flux showed better performance of fluxing a new OSP.

Quality Characteristics of Fermented Beef-Rib Sauce Prepared by Zygosaccharomyces rouxii Cultivation (Zygosaccharomyces rouxii를 배양하여 제조한 발효갈비양념의 품질특성 및 저장성)

  • Kim, Yong-Moon;Oh, Chul-Hwan;In, Man-Jin;Oh, Nam-Soon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.6
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    • pp.807-812
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    • 2007
  • In order to enhance the flavor and storage stability of meat sauce, the manufacturing process of fermented beef-rib sauce was developed in a two-step process. The fermented sauce base was manufactured with Zygosaccharomyces rouxii Y-80 yeast cultivation in raw sauce ingredients for 3 days at $25^{\circ}C$. The fermented beef-rib sauce (FBS) was produced by mixing fermented sauce base with side ingredients. Comparison of the physicochemical and sensory properties with non-fermented beef-rib sauce (NFBS) revealed that the content of ethanol and volatile flavor compounds were higher in FBS; also, the result of sensory evaluation showed that FBS obtained excellent scores for overall taste. To determine the storage stability, FBS and NFBS were incubated at $25^{\circ}C$ for 7 days. The extent of decrease of pH and increase of titratable acidity in NFBS were faster than FBS. After 7 days, ethanol concentrations in FBS and NFBS were 3.77% and 2.04%, respectively. Therefore, based on these results, it can be suggested that storage stability of FBS is superior.

Gene Cloning, Expression and Immunogenicity of the Protective Antigen Subolesin in Dermacentor silvarum

  • Hu, Yonghong;Zeng, Hua;Zhang, Jincheng;Wang, Duo;Li, Dongming;Zhang, Tiantian;Yang, Shujie;Liu, Jingze
    • Parasites, Hosts and Diseases
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    • v.52 no.1
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    • pp.93-97
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    • 2014
  • Subolesin (4D8), the ortholog of insect akirins, is a highly conserved protective antigen and thus has the potential for development of a broad-spectrum vaccine against ticks and mosquitoes. To date, no protective antigens have been characterized nor tested as candidate vaccines against Dermacentor silvarum bites and transmission of associated pathogens. In this study, we cloned the open reading frame (ORF) of D. silvarum 4D8 cDNA (Ds4D8), which consisted of 498 bp encoding 165 amino acid residues. The results of sequence alignments and phylogenetic analysis demonstrated that D. silvarum 4D8 (Ds4D8) is highly conserved showing more than 81% identity of amino acid sequences with those of other hard ticks. Additionally, Ds4D8 containing restriction sites was ligated into the pET-32(a+) expression vector and the recombinant plasmid was transformed into Escherichia coli rosetta. The recombinant Ds4D8 (rDs4D8) was induced by isopropyl ${\beta}$-D-thiogalactopyranoside (IPTG) and purified using Ni affinity chromatography. The SDS-PAGE results showed that the molecular weight of rDs4D8 was 40 kDa, which was consistent with the expected molecular mass considering 22 kDa histidine-tagged thioredoxin (TRX) protein from the expression vector. Western blot results showed that rabbit anti-D. silvarum serum recognized the expressed rDs4D8, suggesting an immune response against rDs4D8. These results provided the basis for developing a candidate vaccine against D. silvarum ticks and transmission of associated pathogens.