• Title/Summary/Keyword: Ala

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Process Strategies to Enhance the Production of 5-Aminolevulinic Acid with Recombinant E. coli

  • LEE , DAE-HEE;JUN, WOO-JIN;YOON, JEONG-WEON;CHO, HONG-YON;HONG, BUM-SHIK
    • Journal of Microbiology and Biotechnology
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    • v.14 no.6
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    • pp.1310-1317
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    • 2004
  • The extracellular production of 5-aminolevulinic acid (ALA) by recombinant E. coli BL21 harboring a fusion gene hemA was investigated in a fermenter. For this purpose, the effects of various physiological factors, such as isopropylthio­$\beta$-D-galactopyranoside (IPTG) concentrations and the time of induction, on enzyme activity were studied. Optimum concentrations of glycine and succinic acid were found to be 30 mM and 90 mM, respectively. When the cells were permitted to grow for 2 h prior to the addition of 0.1 mM IPTG, the activity of ALA synthase was higher than when IPTG was initially added. A 36-fold increase in the activity was observed with only 0.1 mM IPTG added. The pH of the medium also influenced the ALA synthase activity with the maximal activity occurring at pH 6.5. In recombinant E. coli extracts, the repeated addition of glycine and D-glucose increased the production of ALA and the inhibited intracellular ALA dehydratase activity, with up to 32 mM ALA being produced in the cultivation.

Optimization of Influencing Factors on Biomass Accumulation and 5-Aminolevulinic Acid (ALA) Yield in Rhodobacter sphaeroides Wastewater Treatment

  • Liu, Shuli;Li, Xiangkun;Zhang, Guangming;Zhang, Jie
    • Journal of Microbiology and Biotechnology
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    • v.25 no.11
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    • pp.1920-1927
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    • 2015
  • This study aimed to optimize four factors affecting biomass accumulation and 5-aminolevulinic acid (ALA) yield together with pollutants removal in Rhodobacter sphaeroides wastewater treatment. Results showed that it was feasible to produce biomass and ALA in R. sphaeroides wastewater treatment. Microaerobic, 1,000-3,000 lux, and pH 7.0 were optimal conditions for the highest ALA yield of 4.5 ± 0.5 mg/g-biomass. Under these conditions, COD removal and biomass production rate were 93.3 ± 0.9% and 31.8 ± 0.5 mg/l/h, respectively. In addition, trace elements Fe2+, Mg2+, Ni2+, and Zn2+ further improved the ALA yield, COD removal, and biomass production rate. Specifically, the highest ALA yield (12.5 ± 0.6 mg/g-biomass) was achieved with Fe2+ addition.

Facile Synthetic Route to Ascorbic Acid-Dipeptide Conjugate via N-Terminal Activation of Peptide on Resin Support

  • Yang, Jin-Kyoung;Kwak, Seon-Yeong;Jeon, Su-Ji;Kim, Hye-In;Kim, Jong-Ho;Lee, Yoon-Sik
    • Bulletin of the Korean Chemical Society
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    • v.35 no.8
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    • pp.2381-2384
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    • 2014
  • A solid-phase synthetic approach is reported for the synthesis of an ascorbic acid (ASA)-dipeptide conjugate that exhibited enhanced antioxidant activity. The N-terminal amino group of dipeptide (Ala-Ala) on a resin support was first activated by 1,1'-carbonyldiimidazole (CDI), and then reacted with an ASA derivative. The addition of a base, triethylamine (TEA), promoted nucleophilic acylation of ASA derivative and yielded a desired product (ASA-Ala-Ala) with enhanced purity, when cleaved from the resin. Compared to the approach where a C3 hydroxyl group of ASA was first activated with CDI and then reacted with the amino group of dipeptide on the resin, this new approach allowed a significant reduction of a total reaction time from 120 h to 8 h at $25^{\circ}C$. As-prepared ASA-dipeptide conjugate (ASA-Ala-Ala) showed improved antioxidant activity compared to ASA.

Synergistic Anticancer Activity of 5-Aminolevulinic Acid Photodynamic Therapy in Combination with Low-dose Cisplatin on Hela Cells

  • Wei, Xiao-Qiang;Ma, Hui-Qing;Liu, Ai-Hong;Zhang, You-Zhong
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.5
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    • pp.3023-3028
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    • 2013
  • Objective: Photodynamic therapy (PDT ) is a promising modality for the treatment of various tumors. In order to assist in optimizing treatment, we applied 5-ALA/PDT in combination with low-dose cisplatin to evaluate cytotoxicity in Hela cells. Methods: Antiproliferative effects of 5-ALA/PDT and cisplatin, alone and in combination, were assessed using MTT assay. To examine levels of apoptosis, Hela cells treated with 5-ALA/PDT, and combination treatment were assessed with Annexin-V/PI by flow cytometry. To investigate the molecular mechanisms underlying alterations in cell proliferation and apoptosis, Western blot analysis was conducted to determine the expression of p53, p21, Bax and Bcl-2 proteins. Results: MTT assays indicated that combination treatment obviously decreased the viability of Hela cells compared to individual drug treatment. In addition, it was confirmed that exposure of Hela cells to 5-ALA/PDT in combination with low-dose cisplatin resulted in more apoptosis in vitro. Synergistic anticancer activity was related to upregulation p53 expression and alteration in expression of p21, Bcl-2 and Bax. Conclusion: Our findings suggest that administration of 5-ALA/PDT in combination with the low-dose cisplatin may be an effective and feasible therapy for cervical cancer.

Isolation, Purification, and Characterization of the Lytic Enzyme of Anabaena cylindrica by Penicillium oxalicum (HCLF-34) (Penicillium oxalicum(HCLF-34)으로부터 남조세균 (Anabaena cylindrica) 분해효소의 분리 및 동정)

  • 현성희;이호용;최영길
    • Korean Journal of Microbiology
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    • v.36 no.1
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    • pp.14-19
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    • 2000
  • Algal lytic enzyme, an extracellular enzyme, was purified from the culture filtrate of Penicillium oxalicum(HCLF-34) by ultrafiltration, gel filtration chromatography, and anion exchange chromatography. The enzyme has a molecular mass of approximately 22 kDa, an it is a monomer by renaturation SDS-PAGE. The amino acid sequences of the enzyme was revealed to be NH2-Glu-Ser-Tyr-Ser-Ser-Asn-Ala-Ala-Gly-Ala-Val-Leu-Ile---, had about 84% identity with the mature light chain of aspergillopepsin II precursor and 81% identity with the mature protein of the acid proteinase EapC precursor.

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Immunomodulatory Activity of Protein-Bound Polysaccharide Extracted from Gheiidonium majus

  • Yun, Yeon-Sook;Song, Jie-Young;Yang, Hyun-Ok;Pyo, Suhk-Neung;Jung, In-Sung;Yi, Seh-Yoon
    • Archives of Pharmacal Research
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    • v.25 no.2
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    • pp.158-164
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    • 2002
  • In the course of searching immunomodulators from natural sources, the protein-bound polysaccharide, CM-Ala, has been isolated from the water extract of Chelidonium majus L. (Papaveraceae). The immunostimulatory characteristics have been investigated in several experiments such as generation of activated killer (AK) cells, proliferation of splenocytes, activation of macrophages and granulocyte macrophage-colony forming cell (GM-CFC) assay. Of the fractions obtained using Sephacryl S200 column chromatography, CM-Ala was the most effective fraction that augmented the cytotoxicity against Yac-1 tumor cells from 0.88% to 34.18% by culturing with splenocytes for 5 days. CM-Ala also enhanced nitric oxide production by two fold in peritoneal macrophages and exhibited antitumor activity. It showed mitogenic activity on both spleen cells and bone marrow cells. CM-Ala induced proliferation of splenocytes by 84 fold and increased GM-CFC numbers by 1.48 fold over than the non-treated. On the contrary, CM-Ala had cytotoxic activity to a diverse group of tumor cells. From the above results, we proposed that CM-Ala has a possibility of an effective antitumor immunostimulator.

Protective effects of 5-aminolevulinic acid on heat stress in bovine mammary epithelial cells

  • Islam, Md Aminul;Noguchi, Yoko;Taniguchi, Shin;Yonekura, Shinichi
    • Animal Bioscience
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    • v.34 no.6
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    • pp.1006-1013
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    • 2021
  • Objective: Cells have increased susceptibility to activation of apoptosis when suffering heat stress (HS). An effective supplementation strategy to mimic heat-induced apoptosis of bovine mammary epithelial cells (MECs) is necessary to maintain optimal milk production. This study aimed to investigate possible protective effects of the anti-apoptotic activity of 5-aminolevulinic acid (5-ALA) against HS-induced damage of bovine MECs. Methods: Bovine MECs were pretreated with or without 5-ALA at concentrations of 10, 100, and 500 µM for 24 h followed by HS (42.5℃ for 24 h and 48 h). Cell viability was measured with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays. Real-time quantitative polymerase chain reaction and Western blotting were used to explore the regulation of genes associated with apoptosis, oxidative stress, and endoplasmic reticulum (ER) stress genes. Results: We found that 5-ALA induces cytoprotection via inhibition of apoptosis markers after HS-induced damage. Pretreatment of bovine MECs with 5-ALA resulted in dramatic upregulation of mRNA for nuclear factor erythroid-derived 2-like factor 2, heme oxygenase-1, and NAD(P)H quinone oxidoreductase 1, all of which are antioxidant stress genes. Moreover, 5-ALA pretreatment significantly suppressed HS-induced ER stress-associated markers, glucose-regulated protein 78, and C/EBP homologous protein expression levels. Conclusion: 5-ALA can ameliorate the ER stress in heat stressed bovine MEC via enhancing the expression of antioxidant gene.

Dietary Reference Intake of n-3 polyunsaturated fatty acids for Koreans

  • Park, Yongsoon
    • Nutrition Research and Practice
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    • v.16 no.sup1
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    • pp.47-56
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    • 2022
  • This paper examines the process and evidence used to create the Dietary Reference Intake (DRI) of alpha-linolenic acid (ALA) and eicosapentaenoic acid (EPA) + docosahexaenoic acid (DHA) for Koreans. ALA (18:3n3) is an essential fatty acid, and EPA and DHA are known to have beneficial effects on cardiovascular disease risk and reduction of triglyceride levels. Various international organizations have suggested dietary recommendations for n-3 polyunsaturated fatty acids (PUFAs), including ALA, EPA, and DHA. A DRI for Koreans was established for the first time in 2020, specifically for the adequate intake (AI) of ALA and EPA + DHA. This recommendation was based on the average intake of ALA and EPA + DHA from the Korea National Health and Nutrition Examination Survey 2013-2017. For Korean infants, the AI of ALA and DHA was based on the fatty acid composition of maternal milk. Estimated average requirement and a tolerable upper intake level have not been set for n-3 PUFA due to insufficient evidence. In addition, the intake level of n-3 PUFA for prevention of chronic disease has also not been determined. Future studies and randomized controlled trials are required to establish the UL and to define the level for disease prevention.

Performance Evaluation of Artificial Lightweight Aggregate Mortar Manufactured with Waste Glass (폐유리로 제조된 인공경량골재를 이용한 모르타르의 물리적 성능에 대한 평가)

  • Kim, Seong-Soo;Lee, Jeong-Bae;Nam, Ba-Reum;Park, Kwang-Pil
    • Journal of the Korea Concrete Institute
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    • v.21 no.2
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    • pp.147-152
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    • 2009
  • The compressive strength test, bulk density and mortar absorption ratio were carried out to utilize the data as the basic sources for the lightweight mortar and the lightweight concrete, through the study on the physical characteristics of the artificial lightweight aggregate (ALA) made of waste glasses, which was developed for the first time in the country. On the basis of these experiments, the density and the unit volume weight of the ALA showed the value less than 50% of the common aggregate due to the independent pore structure, and the mortar that contains ALA had no big difference from the Control mortar in the test of the absorption ratio. It is judged that this happens based on the internal independent pore structure of the ALA. In case of the mortar containing ALA, there was a tendency of declination in the compressive strength and the bending strength as the mixing rate is increasing, but all mortar showed more than 70% of the Control mortar compressive strength except for the La50 mortar. Hereafter, it is judged that according to the control of the mixing ratio of mineral admixing agent, water and cement, it will realize the equal strength to the control mortar, and the long term edurance is needed to be considered together.

Effects of Dietary L-carnosine and Alpha-lipoic Acid on Growth Performance, Blood Thyroid Hormones and Lipid Profiles in Finishing Pigs

  • Bao, Yinghui;Gao, Chunqi;Hao, Wenbo;Ji, Cheng;Zhao, Lihong;Zhang, Jianyun;Liu, Tao;Ma, Qiugang
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.10
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    • pp.1465-1470
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    • 2015
  • The present study was conducted to determine the effects of L-carnosine (LC) and/or alpha-lipoic acid (ALA) supplementation on growth performance, blood thyroid hormones and lipid profiles in finishing pigs. A total of 40 ($Landrace{\times}Yorkshire$) pigs with an initial body weight of $57.93{\pm}3.14kg$ were randomly allocated to 4 experimental diets using a $2{\times}2$ factorial arrangement with 2 LC supplemental levels (0 or 0.1%) and 2 ALA supplemental levels (0 or 0.03%) in basal diets. The results showed that pigs fed LC-supplemented diets increased final live weight, average daily gain, and average daily feed intake compared to those of pigs fed without LC-supplemented diets (p<0.05). Dietary supplementation with ALA did not affect the growth performance and carcass traits of pigs (p>0.05). Additionally, LC supplementation increased serum triiodothyronine, thyroxine levels, and ALA supplementation increased serum triiodothyronine levels (p<0.05). Serum total cholesterol and triglycerides levels were significantly decreased in LC and ALA supplemented groups, respectively (p<0.05). Moreover, serum low density lipoprotein cholesterol levels were lower in the ALA-supplemented groups than those of pigs fed without ALA-supplemented diets (p<0.05). However, no significant $LC{\times}ALA$ interaction effect on growth performance, blood thyroid hormones and lipid profiles was found. This study suggested that dietary supplementation of LC resulted in better growth performance compared to that of ALA supplementation. L-carnosine and/or ALA supplementation positively modified blood lipid profiles, which may have the potential to prevent cardiovascular diseases.