Yu, Steve Sheng-Fa

Bioorganic Chemistry and Bioinorganic Chemistry

Yu, Steve Sheng-Fa (俞聖法)

Associate Research Fellow (2011-);Assistant Research Fellow (2003-2011);B.S. (1991) and Ph.D. (1998), National Tsing Hua University;Postdoctoral Research Fellow, Institute of Chemistry, Academia Sinica (1999-2003);Joint Assistant Professor, Department of Chemistry, National Cheng Kung University(2005-present);Adjunct Assistant Professor, Department of Materials Science and Engineering, National Tsing Hua University(2005-2011) ;
Research Interests

Development of Fluorinated Mechanistic Probes towards Studies of the Controlled Oxidation of Aliphatics by Metalloenzymes
To initiate the C-H bond oxidation of the aliphatic compounds is a great challenge for synthetic chemists. The conventional synthetic methods and industrial processes are usually required with extremely high temperature and high pressure to carry out the corresponding events. However, some of the gram-negative bacteria, like methanotropic bacteria, Pseudomonas spp. and Bacillus megaterium can facilely convert aliphatic fine chemicals to the corresponding alcohols or oxiranes at ambient temperature and pressure.
gem-Difluoromethylene substituted molecules constitute a distinct class of fluorinated compounds and a special chemistry developed for their preparation. The terminal hydroxylation of these gem-difluorinated octanes by AlkB (Alkane Hydroxylase) from Pseudomonas putida Gpo1 is highly selective for the formation of gem-difluorinated octan-1-ols. The hydroxylation reaction could be performed under conditions of whole-cell catalysis.

Tunning the Reactivity of Metalloproteins for Regio- and Stereoselective Activation of Aliphatics
We employed the water-soluble cytochrome P450 BM-3 to study the activity and regiospecificity of the oxidation on fluorinated n-octanes. By facilitating with appropriate mutations, these variants could regio-selectively convert fluorinated C8 alkanes to their corresponding secondary alcohols. The pattern of reactivity, especially an unprecedented regio-selectivity and stereoselectivity observed for 4,4-difluorooctane, suggests that specific interactions of the fluorinated substituent with aromatic p-systems within the active site can tune the binding of the substrate to the binding pocket as well as the protein conformational states and protein dynamics that participate in electron transfer and C-H activation.

Unraveling the Metal Core Structures of Transcriptional Factors Containing Iron-Sulfur Clusters for their Redox Regulation in Prokaryote
SoxR, a transcriptional factor, is a homodimeric protein with each subunit containing a redox-active [2Fe-2S] center. The iron–sulfur clusters within SoxR regulate transcriptional activity in response to one-electron oxidation, from the paramagnetic mixed-valence [FeII-FeIII] to the diamagnetic [FeIII-FeIII] species; a process that is presumably mediated by oxidative stress. The corresponding oxidation could trigger the expression of the soxS gene and subsequently activate numerous defensive genes, such as superoxide dismutase or catalase in Escherichia coli.
Series of spectroscopic studies of the reduced and oxidized forms of the Escherichia coli transcriptional factor SoxR, followed by mechanistic analyses of the nitrosylation process of these species, allowed the elucidation of the redox chemistry associated with NO sensing that triggers transcriptional activation for the sensing of small molecules in a prokaryote.


細胞色素P-450 BM-3等突變株高專一性地位置與立體選擇性的羥基化反應


Selected Publications
  1. Lo, F.-C.; Lee, J.-F.; Liaw, W.-F.; Hsu, I-J.; Chan, S. I.; Yu, S. S.-F. The Metal Core Structures in the Recombinant Escherichia coli Transcriptional Factor SoxR. Chem. Eur. J., 2012, 18, 2565.
  2. Maji, S.; Lee, J. C.-M.; Lu, Y.-J.; Chen, C.-L.; Hung, M.-C.; Chen, P. P.-Y.; Yu, S. S.-F.; Chan, S. I. Dioxygen Activation of a Trinuclear CuICuICuI Cluster Capable of Mediating Facile Oxidation of Organic Substrates: Competition between O-Atom Transfer and Abortive Inter-cluster Reduction. Chem. Eur. J. 2012, 3955.
  3. Wu, L.-L.; Yang, C.-L.; Lo, F.-C.; Chiang, C.-H.; Chang, C.-W.; Ng, K. Y.; Chou, H.-H.; Hung, H.-Y.; Chan, S. I.; Yu, S. S.-F. Tuning the Regioselectivity and Stereoselectivity in C-H Activation of n-Octanes by Cytochrome P450 BM-3 with Fluorine Substituents.  Evidence for Interactions Between a C-F Bond and Aromatic-pi Systems. Chem. Eur. J. 2011, 17, 4774.
  4. Ramu R.; Chang, C.-W.; Chou, H.-H.; Wu, L.-L.; Chiang, C.-H.; Yu, S. S.-F. Regio-selective Hydroxylation of gem-Difluorinated Octanes by Alkane Hydroxylase (AlkB). Tetrahedron Lett. 2011, 52(23), 2950-3.
  5. Chan S. I.; Nguyen H.-H. T.; Chen, K. H.-C.; Yu, S. S.-F. Over-expression and Purification of the Particulate Methane Monooxygenase from Methylococcus capsulatus (Bath). Methods Enzymol. 2011, 495, 177.
  6. Chan, S. I.; Yu S. S.-F. Controlled Oxidation of Hydrocarbons by the Membrane-bound Methane Monooxygenase: the Case for a Tricopper Cluster. Acc. Chem. Res. 2008, 41(8), 969.
  7. Ng, K.-Y.; Tu, L.-C.; Wang, Y.-S.; Chan, S.-I.; Yu, S. S.-F. Probing the Hydrophobic Pocket of the Active Site in the Particulate Methane Monooxygenase (pMMO) from Methylococcus capsulatus (bath) by Variable Stereoselective Alkane Hydroxylation and Olefin Epoxidation. ChemBioChem 2008, 9, 1116.
  8. Lo, F.-C.; Chen, C.-L.; Lee, C.-M., Tsai, M.-C., Lu, T.-T., Liaw, W.-F., Yu, S. S.-F. A Study of NO Trafficking from Dinitrosyl-iron Complexes to the Recombinant E. coli Transcriptional Factor SoxR. J. Biol. Inorg. Chem. 2008, 13, 961.
  9. Chan, S. I.; Wang, V. C.; Lai, J. C.; Yu, S. S.-F.; Chen, P. P.-Y.; Chen, K. K.-H.; Chen, C.-L.; Chan M.-K. Redox Potentiometry Studies of Particulate Methane Monooxygenase: Support for a Trinuclear Copper Cluster Active Site. Angew. Chem. Int., Ed. 2007, 46(12), 1992.
  10. Yu, S. S. F.; Ji, C.-Z.; Wu, Y.-P.; Lee, T.-L.; Lai, C.-H.; Lin, S.-C.; Yang, Z.-L.; Wang, V. C.-C.; Chen, K. H.-C.; Chan, S. I. The C-terminal Aqueous-exposed Domain of the 45 kDa Subunit of the Particulate Methane Monooxygenase in Methylococcus capsulatus (Bath) is a Cu(I) Sponge. Biochemistry 2007, 46, 13762.
  11. Tseng, C.-F.; Burger, A.; Mislin, G. L. A.; Schalk, I. J.; Yu, S. S.-F.; Chan, S. I.; Abdallah, M. A. Bacterial Siderophores: The Solution Stoichiometry and Coordination of the Fe(III) Complexes of Pyochelin and Related Compounds. J. Biol. Inorg. Chem. 2006, 11, 419.
  12. Chan, S. I.; Chen, K. H.-C.; Yu, S. S.-F.; Chen, C.-L.; Kuo, S. S.-J. Toward Delineating the Structure and Function of the Particulate Methane Monooxygenase (pMMO) from Methanotrophic Bacteria. Biochemistry 2004, 43, 4421.
  13. Yu, S. S.-F.; Chen, H.-C.; Luo,W.-I.; Huang, D.-S.; Chan, S. I. The Stereospecific Hydroxylation of [2,2-2H2]Butaneand Chiral Dideuteriobutanes by the Particulate Methane Monooxygenase from Methylococcus capsulatus (Bath). J. Biol. Chem. 2003, 278, 40658.
  14. Yu, S. S.-F.; Chen, H.-C.; Wang, Y.-S.; Tseng, C. F.; Huang, D.-S.; Chan, S. I. Production of High-Quality Particulate Methane Monooxygenase in High Yields from Methylococcus capsulatus (Bath) with a Hollow-Fiber Membrane Bioreactor. J. Bacteriol. 2003, 185, 5915.

Update: 2015-03-23

Journal Paper
  1. Ramu Ravirala, Wanna Wondemagegn Hailemichael, Janmanchi Damodar, Tsai Yi-Fang, Liu Chih-Cheng, Mou Chung-Yuan, Yu Steve S.-F.* Mechanistic study for the selective oxidation of benzene and toluene catalyzed by Fe(ClO4)2 in an H2O2 -H2O-CH3CN system. Molecular Catalysis 2017-11441, 114-121.
  2. Tsai Yi-Fang, Luo Wen-I, Chang Jen-Lin, Chang Chun-Wei, Chuang Huai-Chun, Ramu Ravirala, Wei Guor-Tzo, Zen Jyh-Myng, Yu Steve S.-F.* Electrochemical Hydroxylation of C3–C12 n-Alkanes by Recombinant Alkane Hydroxylase (AlkB) and Rubredoxin-2 (AlkG) from Pseudomonas putida GPo1. Scientific Reports 2017-087(1), DOI: 10.1038/s41598-017-08610-w.
  3. Wang Vincent C.-C., Maji Suman, Chen Peter P.-Y., Lee Hung Kay, Yu Steve S.-F., Chan Sunney I.* Alkane Oxidation: Methane Monooxygenases, Related Enzymes, and Their Biomimetics. Chemical Reviews 2017-07117(13), 8574-8621.
  4. Yang Chung-Ling, Lin Cheng-Hung, Luo Wen-I, Lee Tsu-Lin, Ramu Ravirala, Ng Kok Yaoh, Tsai Yi-Fang, Wei Guor-Tzo, Yu Steve S.-F.* Mechanistic Study of the Stereoselective Hydroxylation of [2-2 H1 ,3-2 H1 ]Butanes Catalyzed by Cytochrome P450 BM3 Variants. Chemistry - A European Journal 2017-0223(11), 2571-2582.
  5. Chih-Cheng Liu, Ravirala Ramu, Sunney I. Chan*, Chung-Yuan Mou*, Steve S.-F. Yu* Chemistry in confined space: a strategy for selective oxidation of hydrocarbons with high catalytic efficiencies and conversion yields under ambient conditions. Catalysis Science & Technology 2016-09, 6. 7623-7630.
  6. Chih-Cheng Liu, Chung-Yuan Mou, Steve S.-F. Yu, Sunney I. Chan* Heterogeneous formulation of the tricopper complex for efficient catalytic conversion of methane into methanol at ambient temperature and pressure. Energy & Environmental Science 2016-019, 1361-1374.
  7. Minh D. Pham, Ya-Ping Lin, Quan Van Vuong, Penumaka Nagababu,m Brian T.-A. Chang, Kok Yaoh Ng, Chien-Hung Chen, Chau-Chung Han, Chung-Hsuan Chen, Mai Suan Li, Steve S.-F. Yu*, Sunney I. Chan* Inactivation of the particulate methane monooxygenase (pMMO) in Methylococcus capsulatus (Bath) by acetylene.. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS 20151854(12), 1842-52.
  8. Nai-Ti Lin, Kamani Satyanarayana, Chih-Hsien Chen, Yi-Fang Tsai, Steve S.-F. Yu, Sunney I. Chan, Tien-Yau Luh Controlling the Orientation of Pendants in Two-Dimensional Comb-Like Polymers by Varying Stiffness of Polymeric Backbones. MACROMOLECULES 2014-0947(18), 6166-6172.
  9. Yao-Sheng Chen, Wen-I Luo, Chung-Ling Yang, Yi-Jung Tu, Chun-Wei Chang, Chih-Hsiang Chiang, Chi-Yao Chang, Sunney I. Chan, Steve S.-F. Yu* Controlled Oxidation of Aliphatic C-H bonds in Metallo-Monooxygenases: Mechanistic Insights Derived from Studies on Deuterated and Fluorinated Hydrocarbons (Early Career Focused Review, Cover Art). JOURNAL OF INORGANIC BIOCHEMISTRY 2014-05134, 118-133.
  10. Penumaka Nagababu, Steve S.-F. Yu, Suman Maji, Ravirala Ramu and Sunney I. Chan* Developing an efficient catalyst for controlled oxidation of small alkanes under ambient conditions (cover art). CATALYSIS SCIENCE AND TECHNOLOGY 2014-044, 930-935.
  11. Wang DR, Hsiao JC, Wong CH, Li GC, Lin SC, Yu SSF, Chen W, Chang W, Tzou DL Vaccinia Viral Protein A27 is Anchored to the Viral Membrane via a Cooperative Interaction with Viral Membrane Protein A17.. JOURNAL OF BIOLOGICAL CHEMISTRY 2014-03289, 6639-6655.
  12. Peter P.-Y. Chen, Penumaka Nagababu, Steve S.-F. Yu, Sunney I. Chan* Development of the Tricopper Cluster as a Catalyst for the Efficient Conversion of Methane into MeOH. CHEMCATCHEM 2014-026, 429-437.
  13. Yao-Sheng Chen, Wen-I Luo, Tsu-Lin Lee, Steve S.-F. Yu*, Chi-Yao Chang* Identification of the proteins required for fatty acid desaturation in zebrafish (Danio rerio). BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 2013-11440(4), 671-6.
  14. Chih-Hsiang Chiang, Ravirala Ramu, Yi-Jung Tu, Chung-Ling Yang, Kok Yaoh Ng, Wen-I Luo, Charles H. Chen, Yu-Ying Lu, Chen-Lun Liu, Steve S.-F. Yu* Regioselective Hydroxylation of C12-C15 Fatty Acids with Fluorinated Substituents by Cytochrome P450 BM3 (highlighted as a frontispiece of the issue). CHEMISTRY-A EUROPEAN JOURNAL 2013-1019 (41),13680-13691.
  15. Sunney I. Chan*, Yu-Jhang Lu, Penumaka Nagababu, Suman Maji, Mu-Cheng Hung, Marianne M. Lee, I-Jui Hsu, Pham Dinh Minh, Jeff C.-H. Lai, Kok Yoah Ng, Sridevi Ramalingam, Steve S.-F. Yu*, Michael K. Chan* Efficient Oxidation of Methane to Methanol by Dioxygen Mediated by Tricopper Clusters. ANGEWANDTE CHEMIE-INTERNATIONAL EDITION 2013-0352, 3731-3735.
  16. Minh D. Pham, Steve S.-F. Yu, Chau-Chung Han*, Sunney I. Chan*  Improved mass spectrometric analysis of membrane proteins based on rapid and versatile sample preparation on nanodiamond particles.. ANALYTICAL CHEMISTRY 201385(14), 6748-55.
  17. Suman Maji, Jason C.-M. Lee, Yu-Jhang Lu, Chang-Li Chen, Mu-Cheng Hung, Peter P.-Y. Chen, Steve S.-F. Yu, Sunney I. Chan* Dioxygen Activation of a Trinuclear CuICuICuI Cluster Capable of Mediating Facile Oxidation of Organic Substrates: Competition between O-Atom Transfer and Abortive Inter-cluster Reduction. CHEMISTRY-A EUROPEAN JOURNAL 201218(13), 3955-3968.
  18. Penumaka Nagababu, Suman Maji, Manyam Praveen Kumar, Peter, P.-Y. Chen, Steve S.-F Yu*, Sunney I. Chan* Efficient room-temperature oxidation of hydrocarbons mediated by tricopper cluster complexes with different ligands. ADVANCED SYNTHESIS & CATALYSIS 2012354, 3275-3282.
  19. Feng-Chun Lo, Jyh-Fu Lee, Wen-Feng Liaw, I-Jui Hsu*, Yi-Fang Tsai, Sunney I. Chan, Steve S.-F. Yu* The Metal Core Structures in the Recombinant Escherichia coli Transcriptional Factor SoxR. CHEMISTRY-A EUROPEAN JOURNAL 201218(9), 2565-2577.
  20. Ravirala Ramu, Chun-Wei Chang, Ho-Husan Chou, Li-Lan Wu, Chih-Hsiang Chiang, Steve S.-F. Yu* Regio-selective hydroxylation of gem-difluorinated octanes by alkane hydroxylase (AlkB). TETRAHEDRON LETTERS 2011-0652(23), 2950-2953.
  21. Sunney I. Chan*, H.-Hoa T. Nguyen, Kelvin H.-C. Chen, Steve S.-F. Yu* OVER-EXPRESSION AND PURIFICATION OF THE PARTICULATE METHANE MONOOXYGENASE FROM METHYLOCOCCUS CAPSULATUS (BATH). Methods in Enzymology 2011-03495, 177-193.
  22. Li-Lan Wu, Chung-Ling Yang, Feng-Chun Lo, Chih-Hsiang Chiang, Chun-Wei Chang, Kok Yaoh Ng, Ho-Hsuan Chou, Huei-Ying Hung, Sunney I. Chan, Steve S.-F. Yu* Tuning the Regioselectivity and Stereoselectivity in C-H Activation of n-Octanes by Cytochrome P450 BM-3 with Fluorine Substituents. Evidence for Interactions Between a C-F Bond with Aromatic pi-Systems. CHEMISTRY-A EUROPEAN JOURNAL 201117, 4774-4787.
  23. Mansoureh Mousavi, SteveS.-F.Yu, Der-LiiM.Tzou A 13C solid-state NMR analysis of vitamin D compounds. SOLID STATE NUCLEAR MAGNETIC RESONANCE 200936, 24-31.
  24. Shih PC, Yang MS, Lin SC, Ho Y, Hsiao JC, Wang DR, Yu SS, Chang W, Tzou DL A turn-like structure "KKPE" segment mediates the specific binding of viral protein A27 to heparin and heparan sulfate on cell surfaces.. JOURNAL OF BIOLOGICAL CHEMISTRY 2009284(52), 36535-46.
  25. Lo F.-C., Chen C.-L., Lee C.-M., Tsai M.-C., Lu T.-T., Liaw W.-F., Yu S. S.-F.* A study of NO trafficking from dinitrosyl-iron complexes to the recombinant E. coli transcriptional factor SoxR.. JOURNAL OF BIOLOGICAL INORGANIC CHEMISTRY 200813(6), 961-72.
  26. Chan SI, Yu SS Controlled oxidation of hydrocarbons by the membrane-bound methane monooxygenase: the case for a tricopper cluster.. ACCOUNTS OF CHEMICAL RESEARCH 200841(8), 969-79.
  27. Kao WC, Wang VC, Huang YC, Yu SS, Chang TC, Chan SI Isolation, purification and characterization of hemerythrin from Methylococcus capsulatus (Bath).. JOURNAL OF INORGANIC BIOCHEMISTRY 2008102(8), 1607-14.
  28. Ng KY, Tu LC, Wang YS, Chan SI, Yu SS-F* Probing the hydrophobic pocket of the active site in the particulate methane monooxygenase (pMMO) from Methylococcus capsulatus (Bath) by variable stereoselective alkane hydroxylation and olefin epoxidation.. CHEMBIOCHEM 20089(7), 1116-23.
  29. Chan SI, Wang VC, Lai JC, Yu SS, Chen PP, Chen KH, Chen CL, Chan MK Redox potentiometry studies of particulate methane monooxygenase: support for a trinuclear copper cluster active site.. ANGEWANDTE CHEMIE-INTERNATIONAL EDITION 200746(12), 1992-4.
  30. Yu SS, Ji CZ, Wu YP, Lee TL, Lai CH, Lin SC, Yang ZL, Wang VC, Chen KH, Chan SI The C-terminal aqueous-exposed domain of the 45 kDa subunit of the particulate methane monooxygenase in Methylococcus capsulatus (Bath) is a Cu(I) sponge.. BIOCHEMISTRY 200746(48), 13762-74.
  31. Tseng, CF, Burger, A, Mislin, GLA, Schalk, IJ, Yu, SSF, Chan, SI and Abdallah, MA Bacterial siderophores: the solution stoichiometry and coordination of the Fe(III) complexes of pyochelin and related compounds. JOURNAL OF BIOLOGICAL INORGANIC CHEMISTRY 2006-0611(4), 419-432.
  32. M.S. Vinchurkar, K.H.C. Chen, S.S.F. Yu, S.J. Kuo, H.C. Chiu, S.H. Chien and S.I. Chan* Polarized ATR-FTIR spectroscopy of the membrane-embedded domains of the particulate methane monooxygenase.. Biochemistry 200443, 13283-13292.
  33. C.L. Chen, K.H.C. Chen, S.C. Ke, S.S.F. Yu and S.I. Chan* Preparation and characterization of a (Cu, Zn)-pMMO from Methylococcus capsulatus (Bath).. J. Inorg. Biochem. 200498, 2125-2130.
  34. Kao WC, Chen YR, Yi EC, Lee H, Tian Q, Wu KM, Tsai SF, Yu SS, Chen YJ, Aebersold R, Chan SI Quantitative proteomic analysis of metabolic regulation by copper ions in Methylococcus capsulatus (Bath).. J. Biol. Chem. 2004279(49), 51554-60.
  35. S.-C. Hung, C.-Li. Chen, K.H.-C. Chen, S.S.-F. Yu and S.I. Chan* The catalytic copper clusters of the particulate methane monooxygenase from methanotrophic bacteria: Electron paramagnetic resonance spectral simulations.. JOURNAL OF THE CHINESE CHEMICAL SOCIETY 200451, 1229-1244.
  36. K. H.-C. Chen, C.-L. Chen, C.-F. Tseng, S.S.-F. Yu, S.-C. Ke, J.-F. Lee, H.T. Nguyen, S.J. Elliott, J.O. Alben and S.I. Chan* The copper clusters in the particulate methane monooxygenase (pMMO) from Methylococcus capsulatus (Bath) .. JOURNAL OF THE CHINESE CHEMICAL SOCIETY 200451, 1081-1098.
  37. S.I. Chan*, K.H.C. Chen, S.S.F. Yu, C.L. Chen and S.S.J. Kuo Toward delineating the structure and function of the particulate methane monooxygenase from methanotrophic bacteria.. Biochemistry 200443, 4421-4430.
  38. S.S.F. Yu, L.Y. Wu, K.H.C. Chen, M.Y.H. Tseng, Y.S. Wang, C.F. Tseng, Y.J. Chen, D.S. Huang and S.I. Chan* Production of high quality pMMO in high yields from Methylococcus capsulatus (Bath) with a hollow-fiber membrane bioreactor.. J. Bacteriol. 2003185, 5915-5924.
  39. S.S.F. Yu, L.Y. Wu, K.H.C. Chen, W.I. Luo, D.S. Huang, S.I. Chan* The stereospecific hydroxylation of [2,2-2h2]butane and chiral dideuteriobutanes by the particulate methane monooxygenase from Methylococcus capsulatus (Bath).. J. Biol. Chem. 2003278, 40658-40669.
Science Report
  1. 俞聖法、羅鳳君、洪木成、廖文峰、蔡銘哲、魯才德,2009,〈生物系統中的鐵硫簇活性中心蛋白與亞硝基鐵硫錯合物〉,《化學》,第六十七卷第二期,頁133-141。。
  2. 李耀昌、陳俊榮、馮學深、李志甫、黃玉山、李明道、宋豔芳、鄭有舜、江素玉、陳慶曰、黃佩瑜、許益瑞、俞聖法、吳文桂,2008,〈同步輻射在生命科學上的應用與發展〉,《物理雙月刊》,第30卷第1期,頁8-26。
  3. Chan, K. H.-C.; Yu, S. S.-F.; Chan, S. I., 2005, “XAS Studies of pMMO from Methylococcus capsulatus (Bath)”, NSRRC Activity Report, pp. 65.
  1. 2. 陳耀甥、張鈞崴、楊宗霖、吳麗嵐、江志祥、拉姆、羅文一、陳長謙、俞聖法,2013,〈金屬單加氧酵素內其活化烷烴基質選擇性的控制〉,《化學季刊》,第71卷第1期,頁67-81。