• 제목/요약/키워드: co-enzyme regeneration

검색결과 8건 처리시간 0.021초

Photocatalytic Systems of Pt Nanoparticles and Molecular Co Complexes for NADH Regeneration and Enzyme-coupled CO2 Conversion

  • Kim, Ellen;Jeon, Minkyung;Kim, Soojin;Yadav, Paras Nath;Jeong, Kwang-Duk;Kim, Jinheung
    • Rapid Communication in Photoscience
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    • 제2권2호
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    • pp.42-45
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    • 2013
  • Natural photosynthesis utilizes solar energy to convert carbon dioxide and water to energy-rich carbohydrates. Substantial use of sunlight to meet world energy demands requires energy storage in useful fuels via chemical bonds because sunlight is intermittent. Artificial photosynthesis research focuses the fundamental natural process to design solar energy conversion systems. Nicotinamide adenine dinucleotide ($NAD^+$) and $NADP^+$ are ubiquitous as electron transporters in biological systems. Enzymatic, chemical, and electrochemical methods have been reported for NADH regeneration. As photochemical systems, visible light-driven catalytic activity of NADH regeneration was carried out using platinum nanoparticles, molecular rhodium and cobalt complexes in the presence of triethanolamine as a sacrificial electron donor. Pt nanoparticles showed photochemical NADH regeneration activity without additional visible light collector molecules, demonstrating that both photoactivating and catalytic activities exist together in Pt nanoparticles. The NADH regeneration of the Pt nanoparticle system was not interfered with the reduction of $O_2$. Molecular cobalt complexes containing dimethylglyoxime ligands also transfer their hydrides to $NAD^+$ with photoactivation of eosin Y in the presence of TEOA. In this photocatalytic reaction, the $NAD^+$ reduction process competed with a proton reduction.

2단계 고정화 효소반응기를 활용한 Cyclodextrin의 연속생산 (Continuous Production of Cyclodextrin in Two-Stage Immobilized Enzyme Reactor Coupled with Ultrafiltration Recycle System)

  • 이용현;이상호;한일근
    • 한국미생물·생명공학회지
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    • 제19권2호
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    • pp.171-178
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    • 1991
  • The two-stage enzyme reactor, packed with cyclodextrin glucanotransferase (CGTase) immobilized on Amberite IRA 900, coupled with ultrafiltration membrane was investigated for continuous production of cyclodextrin (CD). 5% (w/v) of soluble starch was partially cyclized, in the 0.1 l first-stage immobilized enzyme reactor, up to CD conversion yield of 10% (w/w) at retention time of 0.56hr and 1.5 units of immobilized CGTase/1g of carrier. In the second stage main immobilized enzyme reactor capacity of 1.5 l, the maximum CD conversion yield of 39% (w/v) was achieved at retention time of 2.8hr and 0.47 unit of CGTase/1 g of carrier. Unreacted residual dextrin was fractionated with ultrafiltration membrane, and then, recycled into the second-stage main bioreactor to increase the CD conversion yield. The most suitable membrane size and the volume concentration ratio (concentrate: filterate) for recycling of unreacted residual dextrin were found to be 5K dalton and 4:6, respectively. CD conversion yield was increased about 3~4% upon co-immobilization of pulluanase along with CGTase. Spent Amberite IRA 900 can be reutilized consecutively more than 3 times for immobilization of CGTase after regeneration.

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Improved NADPH Regeneration for Fungal Cytochrome P450 Monooxygenase by Co-Expressing Bacterial Glucose Dehydrogenase in Resting-Cell Biotransformation of Recombinant Yeast

  • Jeon, Hyunwoo;Durairaj, Pradeepraj;Lee, Dowoo;Ahsan, Md Murshidul;Yun, Hyungdon
    • Journal of Microbiology and Biotechnology
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    • 제26권12호
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    • pp.2076-2086
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    • 2016
  • Fungal cytochrome P450 (CYP) enzymes catalyze versatile monooxygenase reactions and play a major role in fungal adaptations owing to their essential roles in the production avoid metabolites critical for pathogenesis, detoxification of xenobiotics, and exploitation avoid substrates. Although fungal CYP-dependent biotransformation for the selective oxidation avoid organic compounds in yeast system is advantageous, it often suffers from a shortage avoid intracellular NADPH. In this study, we aimed to investigate the use of bacterial glucose dehydrogenase (GDH) for the intracellular electron regeneration of fungal CYP monooxygenase in a yeast reconstituted system. The benzoate hydroxylase FoCYP53A19 and its homologous redox partner FoCPR from Fusarium oxysporum were co-expressed with the BsGDH from Bacillus subtilis in Saccharomyces cerevisiae for heterologous expression and biotransformations. We attempted to optimize several bottlenecks concerning the efficiency of fungal CYP-mediated whole-cell-biotransformation to enhance the conversion. The catalytic performance of the intracellular NADPH regeneration system facilitated the hydroxylation of benzoic acid to 4-hydroxybenzoic acid with high conversion in the resting-cell reaction. The FoCYP53A19+FoCPR+BsGDH reconstituted system produced 0.47 mM 4-hydroxybenzoic acid (94% conversion) in the resting-cell biotransformations performed in 50 mM phosphate buffer (pH 6.0) containing 0.5 mM benzoic acid and 0.25% glucose for 24 h at $30^{\circ}C$. The "coupled-enzyme" system can certainly improve the overall performance of NADPH-dependent whole-cell biotransformations in a yeast system.

Efficient Enantioselective Synthesis of (R)-[3,5-Bis(trifluoromethyl)phenyl] Ethanol by Leifsonia xyli CCTCC M 2010241 Using Isopropanol as Co- Substrate

  • Ouyang, Qi;Wang, Pu;Huang, Jin;Cai, Jinbo;He, Junyao
    • Journal of Microbiology and Biotechnology
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    • 제23권3호
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    • pp.343-350
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    • 2013
  • (R)-[3,5-Bis(trifluoromethyl)phenyl] ethanol is a key chiral intermediate for the synthesis of aprepitant. In this paper, an efficient synthetic process for (R)-[3,5- bis(trifluoromethyl)phenyl] ethanol was developed via the asymmetric reduction of 3,5-bis(trifluoromethyl) acetophenone, catalyzed by Leifsonia xyli CCTCC M 2010241 cells using isopropanol as the co-substrate for cofactor recycling. Firstly, the substrate and product solubility and cell membrane permeability of biocatalysts were evaluated with different co-substrate additions into the reaction system, in which isopropanol manifested as the best hydrogen donor of coupled NADH regeneration during the bioreduction of 3,5-bis(trifluoromethyl) acetophenone. Subsequently, the optimization of parameters for the bioreduction were undertaken to improve the effectiveness of the process. The determined efficient reaction system contained 200mM of 3,5-bis(trifluoromethyl) acetophenone, 20% (v/v) of isopropanol, and 300 g/l of wet cells. The bioreduction was executed at $30^{\circ}C$ and 200 rpm for 30 h, and 91.8% of product yield with 99.9% of enantiometric excess (e.e.) was obtained. The established bioreduction reaction system could tolerate higher substrate concentrations of 3,5- bis(trifluoromethyl) acetophenone, and afforded a satisfactory yield and excellent product e.e. for the desired (R)-chiral alcohol, thus providing an alternative to the chemical synthesis of (R)-[3,5-bis(trifluoromethyl)phenyl] ethanol.

Effects of novel chalcone derivatives on α-glucosidase, dipeptidyl peptidase-4, and adipocyte differentiation in vitro

  • Bak, Eun-Jung;Park, Hong-Gyu;Lee, Choong-Hwan;Lee, Tong-Il;Woo, Gye-Hyeong;Na, Young-Hwa;Yoo, Yun-Jung;Cha, Jeong-Heon
    • BMB Reports
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    • 제44권6호
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    • pp.410-414
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    • 2011
  • Chana series are new chalcone derivatives. To evaluate the possibility of Chana series as therapeutic agents of type 2 diabetes, the inhibitory effects of Chana series on the activities of ${\alpha}$-glucosidase and DPP-4 were investigated using in vitro enzyme assays, and their effects on adipocyte differentiation were investigated in C3H10T1/2 cells. Chana 1 and Chana 7 among the Chana series showed significant inhibition of ${\alpha}$-glucosidase activity. In DPP-4 enzyme assay, Chana 1 exhibited the highest inhibitory activity while Chana 7 did not. In MTT assay, Chana 1 did not show significant cytotoxicity up to a concentration of $250{\mu}M$, whereas cytotoxicity was observed with Chana 7 at a concentration of $300{\mu}M$. In addition, Chana 1 induced adipocyte differentiation. Therefore, Chana 1 showed inhibitory effects on ${\alpha}$-glucosidase and DPP-4 as well as a stimulatory effect on adipocyte differentiation, suggesting that Chana 1 may be a potential beneficial agent for the treatment of type 2 diabetes.

핑크-알로에의 항산화, NO 생성 억제 및 세포 재생 효과 (Antioxidant, Inhibitory on NO Production and In-vitro Cell Regeneration Effects of Pink-aloe)

  • 이경주;장욱주;김유아;박병준;강학희
    • 대한화장품학회지
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    • 제46권3호
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    • pp.273-282
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    • 2020
  • 알로에 베라(Aloe barbadensis Miller)는 예로부터 화상이나 상처, 습진 등의 다양한 피부 질환을 개선하기 위해 사용해 왔다. 이러한 효능은 알로에가 가지고 있는 비타민, 무기질, 당, 페놀 물질, 지방산. 아미노산 등의 다양한 성분에 의한 것으로 보고가 되었다. 알로에는 극한의 환경에서 스스로를 보호하기 위해서 성분을 변화시키며, 이로 인해 색깔이 녹색에서 적색으로 변화한다. Aloe-emodin은 알로에속 식물의 대표적인 페놀성 물질로 항산화 및 항염 활성을 나타낸다. 그러나 지금까지 알로에의 활성 물질인 알로에-에모딘을 증가시키고 알로에의 색상을 변화시키는 공정 개발에 대한 연구는 보고되지 않았다. 따라서 본 연구의 목적은 aloe-emodin 함량을 증가시키고 알로에의 항산화 및 항염 활성을 증대시키는 핑크 알로에 제조공정을 개발하는 것이다. 이를 위해 알로에를 적정 조건으로 가온한 결과 핑크 알로에는 일반 알로에 대비 aloe-emodin 함량이 증가하였으며, 항산화 활성 증대, NO 생성 억제, 세포 재생 촉진 등의 효과를 나타내었다. 본 연구를 통해 화장품 분야에서 핑크 알로에의 신규 항노화 소재로 활용성을 확인하였다.

Combination Strategy to Increase Cyclosporin A Productivity by Tolypocladium niveum Using Random Mutagenesis and Protoplast Transformation

  • Lee, Mi-Jin;Duong, Cae Thi Phung;Han, Kyu-Boem;Kim, Eung-Soo
    • Journal of Microbiology and Biotechnology
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    • 제19권9호
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    • pp.869-872
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    • 2009
  • The cyclic undecapeptide cyclosporin A (CyA), one of the most valuable immunosuppressive drugs, is produced nonribosomally by a multifunctional cyclosporin synthetase enzyme complex by the filamentous fungus Tolypocladium niveum. To increase CyA productivity by wild-type T. niveum (ATCC 34921), random mutagenesis was first performed using an antifungal agar-plug colony assay (APCA) selection approach. This generated a mutant strain producing more than 9-fold greater CyA than the wild-type strain. Additionally, a foreign bacterial gene, Vitreoscilla hemoglobin gene (VHb), was transformed via protoplast regeneration and its transcription was confirmed by RT-PCR in the UV-irradiated mutant cell. This led to an additional 33.5% increase of CyA production. Although most protoplast-regenerated T. niveum transformants tend to lose CyA productivity, the optimized combination of random mutagenesis and protoplast transformation described here should be an efficient strategy to generate a commercially valuable, yet metabolite low-producing, fungal species, such as CyA-producing T. niveum.

손상된 불가사리 추출물 흡입이 흡연으로 인한 항염증과 항산화 효소의 효과 (The Injured Starfish Extract Inhalation Effects of Anti-inflammation and Anti-oxidants Enzyme during Indirect Cigarette Smoking)

  • 황경희;정혁;장수찬;박종석;김유영
    • KSBB Journal
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    • 제27권6호
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    • pp.367-374
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    • 2012
  • Cigarette smoking (SM) is considered to be well known environmental toxin which contributes to the onset of various diseases. SM cause direct lungs damage, activate lungs inflammatory responses, and in some cases leads to the development of lung cancer. Cytokines in injured starfish (Asterina pectinifera) is the potential changes in its expression during the regeneration process. Especially, expression of TGF-${\beta}1$ has increased in arm cut starfish extract after eight days. Also, starfish including saponin like the ginseng. Saponin is widely used in the world because of some effective pharmacological activities. Therefore, the current study was designed to elucidate the pharmacological activities of starfish extract against cigarette smoking induced damage in cell line and pulmonary tissue. We investigate that the effect of eight days starfish extract after arm cut (8d) and intact starfish extract on cell line and mouse lung injury by SM. In cell proliferation analysis, although cigarette smoking extract (CSE) was co-treated, the higher proliferation ability is shown in 8d treatment than intact starfish extract. 8d and intact starfish extract was directly transported to pulmonary cells through respiratory organ by nebulizer inhalation. In this case of cigarette smoking, the pulmonary structure was damaged and functions become abnormal. However, 8d treated groups showed similar with the control group compared with SM group. Among them, 8d was proved to be more effective than intact starfish extract. These results demonstrate that 8d could more protect pulmonary structure and function than intact starfish extract against cigarette smoking by ginseng like saponin and regulation of inflammatory cytokines.