Browsing by Author "Wu, Tung-Kung"

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A Ruthenium-Based Biomimetic Hydrogen Cluster for Efficient Photocatalytic Hydrogen Generation from Formic Acid
(2015-04-20)
Chang, Chin-Hao
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Chen, Mei-Hua
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Du, Wan-Shan
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Gliniak, Jacek
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Lin, Jia-Hoa
A ruthenium-based biomimetic hydrogen cluster, [Ru-2(CO)(6)(-SCH2CH2CH2S)] (1), has been synthesized and, in the presence of the P ligand tri(o-tolyl)phosphine, demonstrated efficient photocatalytic hydrogen generation from formic acid decomposition. Turnover frequencies (TOFs) of 5500h(-1) and turnover numbers (TONs) over 24700 were obtained with less than 50ppm of the catalyst, thus representing the highest TOFs for ruthenium complexes as well as the best efficiency for photocatalytic hydrogen production from formic acid. Moreover, 1 showed high stability with no significant degradation of the photocatalyst observed after prolonged photoirradiation at 90 degrees C.
AC electrokinetics assisted impedance biosensors for rapid bacteria detection
(2012)
Ho, Yi-Tse
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Pan, Chun-Liang
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Sheu, Jeng-Tzong
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Wang, Yu-Kuo
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Wu, Tung-Kung
Early detection of living bacteria attracts a lot of attention in infection diseases applications, especially at low concentration. Nowadays, the detection of specific bacteria faces challenges including enhancement of trapping efficiency, and increase of sensor sensitivity and specificity. In this study, we improved the trapping efficiency and increased sensor sensitivity and specificity by using AC electrokinetics and selective modification of antibody on gold electrode in a new structure of gold electrode design. COMSOL simulations of AC electrokinetics include dielectrophoresis (DEP), and AC electroosmosis (ACEO) were performed on different electrode designs and bias configurations. The simulation results were consistent with the fluorescence observations showing that the bacteria were trapping selectively on gold electrodes. The impedance measurements for Vibrio parahaemolyticus (Vp) detection at 10(5) cfu/ml with AC elctrokinetics showed a nearly 90% change in normalized impedance change (NIC).
Alteration of the Substrate's Prefolded Conformation and Cyclization Stereochemistry of Oxidosqualene-Lanosterol Cyclase of Saccharomyces cerevisiae by Substitution at Phenylalanine 699
(2010-02-05)
Wu, Tung-Kung
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Chang, Cheng-Hsiang
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Wen, Hao-Yu
;
Liu, Yuan-Ting
;
Li, Wen-Hsuan
The Saccharomyces cerevisiae ERG7(Phe699) mutants produced one chair-chair-chair (C-C-C) and two chair-boat-chair (C-B-C) truncated tricyclic compounds, one tetracyclic 17 alpha-exocyclic unrearranged intermediate, and two 17 beta-exocyclic truncated rearranged intermediates. These results provided direct evidence for the importance of the residue in affecting mechanistic transitions between C-B-C and C-C-C substrate conformation and between the 17 alpha- and 17 beta-exocyclic side chain stereochemistry as well as in stabilizing the 6 -6-5 tricyclic and the protosteryl C-17 cations.
Altering oxidosqualene cyclization from chair-boat-chair to chair-chair-chair biosynthetic route by a single amino acid substitution at the phenylalanine 699 position of the Saccharomyces cerevisiae oxidosqualene-lanosterol cyclase
(2009-08-16)
Wu, Tung-Kung
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應用化學系分子科學碩博班
;
Institute of Molecular science
An enzymatic approach to configurationally rare trans-androsteronyl--glucoside and Its potential anticancerapplication
(2017-01)
Chou, Feng-Pai
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Tsai, Chia-Tse
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Chiou, Ya-Sheng
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Chen, Yi-Ju
;
Li, Meng-Erh
Enzymatic glycosylation of sterols/steroids with glycosyltransferase HP0421 shows protein plasticity on generation of configurationally rare steryl--glucosides. Investigation of trans-androsteronyl--glucoside on tamoxifen-treated MCF-7 breast cancer cells shows dose-dependent depression of cell viability and enhanced drug effectiveness, illustrating a new avenue for the production of novel steryl--glucosides with useful biological activities.
Anticancer and Antioxidant Activities of the Peptide Fraction from Algae Protein Waste
(2010-01-27)
Sheih, I-Chuan
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Fang, Tony J.
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Wu, Tung-Kung
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Lin, Peng-Hsiang
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生物科技學系
Algae protein waste is a byproduct during production of algae essence from Chlorella vulgaris. There is no known report on the anticancer peptides derived from the microalgae protein waste. In this paper, the peptide fraction isolated from pepsin hydrolysate of algae protein waste had strong dose-dependent antiproliferation and induced a post-G1 cell cycle arrest in AGS cells; however, no cytotoxicity was observed in WI-38 lung fibroblasts cells in vitro. The peptide fraction also revealed much better antioxidant activity toward peroxyl radicals and LDL than those of Trolox. Among these peptides, a potent antiproliferative, antioxidant, and NO-production-inhibiting hendecapeptide was isolated, and its amino acid sequence was VECYGPNRPQF. These results demonstrate that inexpensive algae protein waste could be a new alternative to produce anticancer peptides.
Antioxidant properties of a new antioxidative peptide from algae protein waste hydrolysate in different oxidation systems
(2009-07-01)
Sheih, I. -Chuan
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Wu, Tung-Kung
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Fang, Tony J.
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生物科技學系
;
Department of Biological Science and Technology
Microalgae have been a popular edible food, but there are no known reports on the antioxidative peptides derived from microalgae. The algae protein waste, which is normally discarded as animal feed, is a by-product during production of algae essence from microalgae, Chlorella vulgaris. Algae protein waste was hydrolyzed using pepsin, and a potent antioxidative peptide of VECYGPNRPQF was separated and isolated. The peptide could efficiently quench a variety of free radicals, including hydroxyl radical, superoxide radical, peroxyl radical, DPPH radical and ABTS radicals. and performed more efficiently than that observed for BHT, Trolox and peptides from marine protein sources in most cases. The purified peptide also has significant protective effects on DNA and prevents cellular damage caused by hydroxyl radicals. In addition, the peptide has gastrointestinal enzyme-resistance and no cytotoxicity observed in human lung fibroblasts cell lines (WI-38) in vitro. These results demonstrate that inexpensive algae protein waste could be a new alternative to produce antioxidative peptides. (C) 2009 Elsevier Ltd. All rights reserved.
BIOT 373-Artificial myoglobin system design for electron transfer mechanism study and BioSolar cell applications
(2006-09-10)
Chang, Chin-Hao
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Wu, Tung-Kung
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Diau, Eric Wei Guang
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Luo, Li-Yang
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生物科技學系
Characterization and epitope mapping of Dengue virus type 1 specific monoclonal antibodies
(2017-10-02)
Chen, Wen-Hung
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Chou, Feng-Pai
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Wang, Yu-Kuo
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Huang, Sheng-Cih
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Cheng, Chuan-Hung
Background: Dengue virus (DV) infection causes a spectrum of clinical diseases ranging from dengue fever to a life-threatening dengue hemorrhagic fever. Four distinct serotypes (DV1-4), which have similar genome sequences and envelope protein (E protein) antigenic properties, were divided. Among these 4 serotypes, DV1 usually causes predominant infections and fast diagnosis and effective treatments are urgently required to prevent further hospitalization and casualties. Methods: To develop antibodies specifically targeting and neutralizing DV1, we immunized mice with UV-inactivated DV1 viral particles and recombinant DV1 E protein from residue 1 to 395 (E395), and then generated 12 anti-E monoclonal antibodies (mAbs) as the candidates for a series of characterized assays such as ELISA, dot blot, immunofluorescence assay, western blot, and foci forming analyses. Results: Among the mAbs, 10 out of 12 showed cross-reactivity to four DV serotypes as well as Japanese encephalitis virus (JEV) in different cross-reactivity patterns. Two particular mAbs, DV1-E1 and DV1-E2, exhibited strong binding specificity and neutralizing activity against DV1 and showed no cross-reactivity to DV2, DV3, DV4 or JEV-infected cells as characterized by ELISA, dot blot, immunofluorescence assay, western blot, and foci forming analyses. Using peptide coated indirect ELISA, we localized the neutralizing determinants of the strongly inhibitory mAbs to a sequence-unique epitope on the later-ridge of domain III of the DV1 E protein, centered near residues T346 and D360 ((346)TQNGRLITANPIVTD(360)). Interestingly, the amino acid sequence of the epitope region is highly conserved among different genotypes of DV1 but diverse from DV2, DV3, DV4 serotypes and other flaviviruses. Conclusions: Our results showed two selected mAbs DV1-E1 and DV1-E2 can specifically target and significantly neutralize DV1. With further research these two mAbs might be applied in the development of DV1 specific serologic diagnosis and used as a feasible treatment option for DV1 infection. The identification of DV1 mAbs epitope with key residues can also provide vital information for vaccine design.
Crystal Structure and Mutational Analysis of Aminoacylhistidine Dipeptidase from Vibrio alginolyticus Reveal a New Architecture of M20 Metallopeptidases
(2010-12-01)
Chang, Chin-Yuan
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Hsieh, Yin-Cheng
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Wang, Ting-Yi
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Chen, Yi-Chin
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Wang, Yu-Kuo
Aminoacylhistidine dipeptidases (PepD, EC 3.4.13.3) belong to the family of M20 metallopeptidases from the metallopeptidase H clan that catalyze a broad range of dipeptide and tripeptide substrates, including L-carnosine and L-homocarnosine. Homocarnosine has been suggested as a precursor for the neurotransmitter gamma-aminobutyric acid (GABA) and may mediate the antiseizure effects of GABAergic therapies. Here, we report the crystal structure of PepD from Vibrio alginolyticus and the results of mutational analysis of substrate-binding residues in the C-terminal as well as substrate specificity of the PepD catalytic domain-alone truncated protein PepD(CAT). The structure of PepD was found to exist as a homodimer, in which each monomer comprises a catalytic domain containing two zinc ions at the active site center for its hydrolytic function and a lid domain utilizing hydrogen bonds between helices to form the dimer interface. Although the PepD is structurally similar to PepV, which exists as a monomer, putative substrate-binding residues reside in different topological regions of the polypeptide chain. In addition, the lid domain of the PepD contains an "extra" domain not observed in related M20 family metallopeptidases with a dimeric structure. Mutational assays confirmed both the putative di-zinc allocations and the architecture of substrate recognition. In addition, the catalytic domain-alone truncated PepD(CAT) exhibited substrate specificity to L-homocarnosine compared with that of the wild-type PepD, indicating a potential value in applications of PepD(CAT) for GABAergic therapies or neuroprotection.
Crystal Structure and Mutational Analysis of Aminoacylhistidine Dipeptidase from Vibrio alginolyticus Reveal a New Architecture of M20 Metallopeptidases
(2010-12-01)
Chang, Chin-Yuan
;
Hsieh, Yin-Cheng
;
Wang, Ting-Yi
;
Chen, Yi-Chin
;
Wang, Yu-Kuo
Aminoacylhistidine dipeptidases (PepD, EC 3.4.13.3) belong to the family of M20 metallopeptidases from the metallopeptidase H clan that catalyze a broad range of dipeptide and tripeptide substrates, including L-carnosine and L-homocarnosine. Homocarnosine has been suggested as a precursor for the neurotransmitter gamma-aminobutyric acid (GABA) and may mediate the antiseizure effects of GABAergic therapies. Here, we report the crystal structure of PepD from Vibrio alginolyticus and the results of mutational analysis of substrate-binding residues in the C-terminal as well as substrate specificity of the PepD catalytic domain-alone truncated protein PepD(CAT). The structure of PepD was found to exist as a homodimer, in which each monomer comprises a catalytic domain containing two zinc ions at the active site center for its hydrolytic function and a lid domain utilizing hydrogen bonds between helices to form the dimer interface. Although the PepD is structurally similar to PepV, which exists as a monomer, putative substrate-binding residues reside in different topological regions of the polypeptide chain. In addition, the lid domain of the PepD contains an "extra" domain not observed in related M20 family metallopeptidases with a dimeric structure. Mutational assays confirmed both the putative di-zinc allocations and the architecture of substrate recognition. In addition, the catalytic domain-alone truncated PepD(CAT) exhibited substrate specificity to L-homocarnosine compared with that of the wild-type PepD, indicating a potential value in applications of PepD(CAT) for GABAergic therapies or neuroprotection.
Crystal structure of aminoacylhistidine dipeptidase from vibrio alginolyticus, a metallopeptidase with dinuclear metal center.
(2009-08-16)
Chang, Chin Yuan
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Wu, Tung-Kung
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生物科技學系
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Department of Biological Science and Technology
Differential Stereocontrolled Formation of Tricyclic Triterpenes by Mutation of Tyrosine 99 of the Oxidosqualene-Lanosterol Cyclase from Saccharomyces cerevisiae
(2009-11-01)
Wu, Tung-Kung
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Li, Wen-Hsuan
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Chang, Cheng-Hsiang
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Wen, Hao-Yu
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Liu, Yuan-Ting
The function of the Tyr99 residue from Saccharomyces cerevisiae oxidosqualene-lanosterol cyclase (ERG7) was analyzed by constructing deletion and site-saturated mutants. Two truncated intermediates, (13 alpha H)-isomalabarica-14Z,17E,21-trien-3 beta-ol and (13 alpha H)-isomalabarica-14E,17E,21-trien-3 beta-ol, were isolated from the ERG7(Y99X) mutants. These results suggest that the functional role of ERG7(Y99) is to affect both chair-boat 6-6-5 tricyclic Markovnikov C-14 cation stabilization and the stereochemistry of the protons at the C-15 position for subsequent deprotonation. ((C) Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009)
Effects of fermentation on antioxidant properties and phytochemical composition of soy germ
(2014-12-01)
Sheih, I-Chuan
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Fang, Tony J.
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Wu, Tung-Kung
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Chen, Ru-Yin
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生物科技學系
BACKGROUNDTraditional soy-fermented foods, such as miso, douche, natto, and tempeh have been widely used as a dietary supplement in Asian countries, and numerous reports on their phenolics and antioxidant activities have been published. Soy germ contains 10-fold higher phenolics than whole soybean, hence using soy germ as fermentation substrate will be more efficient than whole soybean. RESULTSSoy germ fermented with Aspergillus nigerM46 resulted in a high-efficiency bio-transformation of phenolics and flavonoids to their metabolites, and a diverse secondary metabolic product was also found to response oxidation stress of fungal colonisation. Its antioxidant activity against hydroxyl radicals and superoxide radicals (IC50 = 0.8 and 6.15 mu g mL(-1), respectively) was about 205-fold and 47-fold higher than those of unfermented soy germ (IC50 = 164.0 and 290.48 mu g mL(-1)), respectively. These results were similar to those observed for Trolox, and more active than those of BHT and hesperidin. The -glucosidase and -amylase produced during fermentation were mainly responsible for mobilisation of the phenolics. CONCLUSIONOur results demonstrate that fermented soy germ has the potential to be a good dietary supplement for prevention of oxidative stress-related diseases, and the solid-state bioprocessing strategy could be an innovative approach to enhance the antioxidant activity of soy germ. (c) 2014 Society of Chemical Industry
Engineering Fungal Nonreducing Polyketide Synthase by Heterologous Expression and Domain Swapping
(2013-02-15)
Yeh, Hsu-Hua
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Chang, Shu-Lin
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Chiang, Yi-Ming
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Bruno, Kenneth S.
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Oakley, Berl R.
We reannotated the A. niger NR-PKS gene, e_gw1_19.204, and its downstream R domain gene, est_GWPlus_C_190476, as a single gene which we named dtbA. Heterologous expression of dtbA In A. nidulans demonstrated that DtbA protein produces two polyketides, 2,4-dihydroxy-3,5,6-trimethylbenzaldehyde (1) and 6-ethyl-2,4-dihydroxy-3,5-dimethylbenzaldehyde (2). Generation of DtbA Delta R+TE chimeric PKSs by swapping the DtbA R domain with the AusA (austinol biosynthesis) or ANID_06448 TE domain enabled the production of two metabolites with carboxylic acids replacing the corresponding aldehydes.
Enhanced hydrogen generation from formic acid by a biomimetic ruthenium complex with a covalently bonded phosphine ligand
(2017-04-13)
Chen, Mei-Hua
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Lin, Jia-Hoa
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Li, Chuen-Ru
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Gliniak, Jacek
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Chang, Chin-Hao
A series of [Ru-2(CO)(5)(mu-SCH2CH2CH2S)PXS] complexes (Ru-2-S-2-PXS, X = phosphine ligands, S = 1-8) have been synthesized and evaluated for their photocatalytic H-2 generation efficiencies from formic acid decomposition. The [Ru-2(CO)(5)(mu-SCH2CH2CH2S)P(o-C6H4CH3)(3)] (Ru-2-S-2-PX4) catalyst + P(CH3)(3) ligand exhibited a high turnover frequency of 15,840h(-1) and turnover number of 24,536. A mechanistic investigation of the Ru-2-S-2-PX4 + FA/TEA catalyzed photocatalytic H-2 generation reaction using ATR-IR, EI-MS, and NMR techniques suggested that when Ru2-S2-PX4 was photoirradiated, the P(o-C6H4CH3)(3) was dissociated from the complex to form a new species, [Ru-2(CO)(5)(mu-SCH2CH2CH2S)]* (I). The free P(o-C6H4CH3)(3) then attacks a second molecule of Ru-2-S-2-PX4 to form Ru-2-S-2-(PX4)(2) and release of free CO, which is then combined with species I to form Ru-2-S-2. Subsequent attachment of formate ion to species Ru-2-S-2-PX4, Ru-2-S-2, and Ru-2-S-2-(PX4)(2) to form [Ru-2(CO)(5)(mu-SCH(2)CH(2)CH(2)SA]-HC00(-) (II), Ru-2-S-2-HCOO- (II') and Ru-2-S-2-PX4-HCOO- (II"), respectively. Rearrangement of complex II (or II' or II") and evolution of CO2 generate a transient complex [Ru-2(CO)(5)(mu-SCH2CH2CH2S)H (III), which then undergoes a protonation process to yield complex [Ru-2(CO)(5)(mu-SCH2CH2CH2S)H-2] (IV). Release of H-2 and re-incorporation of the formate anion as well as evolution of CO2 regenerates the active complex III and the cycle begins again. (C) 2017 Hydrogen Energy Publications LLC. Published by Elsevier Ltd. All rights reserved.
Enzymatic approach to configurationally rare trans-androsteronyl-alpha-glucoside and its potential for anticancer application
(2016-07-25)
Wu, Tung-Kung
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交大名義發表
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National Chiao Tung University
Enzymatic approach to configurationally rare trans-androsteronyl-alpha-glucoside and its potential for anticancer application
(2016-07-25)
Wu, Tung-Kung
;
交大名義發表
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National Chiao Tung University
Expression and characterization of the biofilm-related and carnosine-hydrolyzing aminoacylhistidine dipeptidase from Vibrio alginolyticus
(2008-10-01)
Wang, Ting-Yi
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Chen, Yi-Chin
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Kao, Liang-Wei
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Chang, Chin-Yuan
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Wang, Yu-Kuo
The biofilm-related and carnosine-hydrolyzing aminoacylhistidine dipeptidase (pepD) gene from Vibrio alginolyticus was cloned and sequenced. The recombinant PepD protein was produced and biochemically characterized and the putative active-site residues responsible for metal binding and catalysis were identified. The recombinant enzyme, which was identified as a homodimeric dipeptidase in solution, exhibited broad substrate specificity for Xaa-His and His-Xaa dipeptides, with the highest activity for the His-His dipeptide. Sequence and structural homologies suggest that the enzyme is a member of the metal-dependent metallopeptidase family. Indeed, the purified enzyme contains two zinc ions per monomer. Reconstitution of His.Tag-cleaved native apo-PepD with various metal ions indicated that enzymatic activity could be optimally restored when Zn(2+) was replaced with other divalent metal ions, including Mn(2+), Co(2+), Ni(2+), Cu(2+) and Cd(2+), and partially restored when Zn(2+) was replaced with Mg(2+). Structural homology modeling of PepD also revealed a 'catalytic domain' and a 'lid domain' similar to those of the Lactobacillus delbrueckii PepV protein. Mutational analysis of the putative active-site residues supported the involvement of His80, Asp119, Glu150, Asp173 and His461 in metal binding and Asp82 and Glu149 in catalysis. In addition, individual substitution of Glu149 and Glu150 with aspartic acid resulted in the partial retention of enzymatic activity, indicating a functional role for these residues on the catalysis and zinc ions, respectively. These effects may be necessary either for the activation of the catalytic water molecule or for the stabilization of the substrate-enzyme tetrahedral intermediate. Taken together, these results may facilitate the design of PepD inhibitors for application in antimicrobial treatment and antibody-directed enzyme prodrug therapy.
Fabrication and Photovoltaic Characterization of Bio-Sensitized Solar Cells Using Myoglobin-Based Sensitizers
(2009-03-01)
Chang, Chih-Wei
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Chang, Chin-Hao
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Lu, Hsueh-Pei
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Wu, Tung-Kung
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Diau, Eric Wei-Guang
Myoglobin (Mb), reconstituted zinc protoporphyrin-apornyoglobin (ZnMb), and eosin-modified ZnMb (EoZnMb) were used as photosensitizers to functionalize TiO(2) nanocrystalline films for biosensitized solar-cell (BSSC) applications. For the Mb-sensitized SC, the poor cell performance was due to a reduction Fe(III) -> Fe(II) that produces a photocurrent density of the device smaller than its unsensitized counterpart. The efficiencies of power conversion of both ZnMb and EoZnMb-sensitized SC were enhanced about ten times due to superior charge separation between TiO(2) and the protein, and due to smaller current leakage between TiO(2) and the electrolyte. The cell performances of the BSSC devices are discussed in terms of an equivalent-circuit model.