生物催化与分子进化
 

　　1、高效高选择性功能酶的筛选、表达和分子改造；  

　　2、新型多酶生物催化体系及化学-酶偶联体系的设计和构建； 

　　3、高值手性化合物的酶催化合成新方法和新工艺研究；  

　　4、生物催化法构建多样性的活性分子库和新型类药分子。  

　　  

　　实验室构建了高效、高选择性的酶库和工具箱，主要涉及环氧化酶、细胞色素P450酶、羰基还原酶，烯烃还原酶等多种氧化还原酶类；构建了成熟的高通量筛选平台和蛋白质工程改造技术平台，开展酶蛋白及突变体的结构功能关系研究，实现生物催化剂的设计定制。 

　　

招收有机化学、酶工程、分子生物学、微生物学等相关学科硕士/博士研究生

081703-生物化工
071005-微生物学

生物催化
蛋白质工程
绿色制造

1998-09--2003-06   中国科学院上海有机化学研究所   理学博士学位
1994-09--1998-06   华东理工大学   理学学士学位

-- 研究生

-- 博士

生物催化基础与应用

1.     Zhu, B.-F.; M. Qaiser Fatmi, Pei, X.-Q.; Zhong-Liu Wu,* Liu, Y.* Continuous Engineering of Phenylalanine Ammonia Lyase from Lettuce (Lactuca sativa L.) for Efficient Synthesis of 3,4-Substituted Phenylalanine. J. Agric. Food Chem. 2025, 73, 17, 10358–10368

2.     Li, Z.-P.; Wang, L.; Liu, Y.; Pei, X.-Q.; Fatmi, M. Q.; Shen, Z.; Zhao, J.; Lin, H. *; Zhou, J. *; Wu, Z.-L*. Structural and Mechanistic Insight into the Enantioselectivity of (R)-Selective Styrene Monooxygenases: A Tug-of-War between Proximal and Distal Residues. Angew. Chem. Int. Ed. 2025, 64, e202423117.

3.     Qiang Chen, Zhi-Pu Li, Xiao-Qiong Pei, Yan Liu, M. Qaiser Fatmi*, Zhong-Liu Wu,* Thermostabilization and molecular dynamics simulation of NaStyA for highly enantioselective (R)-epoxidation of styrene analogues. Molecular Catalysis 564 (2024)114312

4.     Shi, M.-L.; Lin, R.-D.; Yu, Y.; Yao, Y.; Fan, X.-Y; Li, K.; Wu, Z.-L.; Yu, X.-Q.; Liu, Y.*; Wang. N.*. Efficient Stereoselective Synthesis of Aryl Lactones Using Engineered Ketoreductase ChKRED20 from Chryseobacterium sp. CA49. Organic Chemistry Frontiers. Org. Chem. Front., 2024, DOI: 10.1039/D3QO02076B. 11 (6), pp.1804-1810

5.     Lin, H.; Fang, S. Y.; Gao, H.; Qin, Z. Y.; Fan, D. L.; Li, N.; Wu, Z. L.; Chen, H. G., Mechanistic Insights into Enantiocontrolling of Styrene Monooxygenase-Catalyzed Epoxidation of Olefins. ACS Catal 2024, 14 (4), 2246-2251.

6.     Chen, Q.; Xiao, H.; Li, Z.-P.; Pei, X.-Q.; Yang, W.; Liu, Y.*; Wu, Z.-L.* Stereo-complementary epoxidation of 4-vinyl-2,3-dihydrobenzofuran using mutants of SeStyA with enhanced stability and enantioselectivity. Molecular Catalysis, 2023, 540: 113055.

7.     Zhu, B.-F.; Liu, Y.*; Pei, X.-Q.; Wu, Z.-L.* Characterization of Phenylalanine Ammonia Lyases from Lettuce (Lactuca sativa L.) as Robust Biocatalysts for the Production of D and L Amino Acids. J. Agric. Food Chem. 2023, 71 (6):2935-2942.

8.     Xing, Xiu; Liu, Yan; Lin, Ru-De; Zhang, Yang; Wu, Zhong-Liu; Yu, Xiao-Qi; Li, Kun; Wang, Na. Development of an integrated system for highly selective photoenzymatic synthesis of formic acid from CO2. Chemsuschem, 2023, 16(5):e202201956; DOI:10.1002/cssc.202201956.

9.     Liu, Y.; Chen, Q.; Zhu, B. F.; Pei, X.-Q.; Liu, Y.; Wu, Z. L.*. Sequence-guided stereo-enhancing and -inverting of (R)-styrene monooxygenases for highly enantioselective epoxidation. Molecular Catalysis 531 (2022) 112680

10.   Liu, Y.-C.; Wu, Z. -L.; Deska, J. Coding Synthetic Chemistry Strategies for Furan Valorization into Bacterial Designer Cells. ChemSusChem. 2023, 16(2): e202201790. DOI10.1002/cssc.202201790.

11.   Lu, W.-F.; Yu, Y., Lin, R. D.; Yao, Y.; Liu, Y.; Wu, Z. L.; Liu, Y. ; Wang, N.* Enantioselective biosynthesis of (R)-γ-hydroxy sulfides via a one-pot approach with ChKRED20. Molecular Catalysis 531(2022)112655.

12.   Lin, H.*; Meng, Y.; Li, N.; Tang, Y.; Dong, S.; Wu, Z.; Xu, C.; Kazlauskas, R.*; Chen, H.*, Enzymatic Enantioselective anti-Markovnikov Hydration of Aryl Alkenes. Angew Chem Int Ed Engl 2022, 61 (32), e202206472.

13.   Yang, Y. J.; Pei, X. Q.; Liu, Y.; Wu, Z. L. *, Thermostabilizing ketoreductase ChKRED20 by consensus mutagenesis at dimeric interfaces. Enzyme Microb Technol 2022, 158, 110052.

14.   Xing, X.; Liu, Y.*; Shi, M.-L.; Li, K.; Fan, X.-Y.; Wu, Z.-L.; Wang, N.*; Yu, X.-Q. Preparation of chiral aryl alcohols: a controllable enzymatic strategy via light-driven NAD(P)H regeneration. New J. Chem. 2022, 46 (13), 6274-6282.

15.   Xiao, H.; Dong, S.; Liu, Y.; Pei, X.-Q.; Lin, H.*; Wu, Z.-L.* A new clade of styrene monooxygenases for (R)- selective epoxidation. Catalysis Science & Technology, 2021,11: 2195–2201.

16.   Liu, Y.*; Li, Z.-Y.; Guo, C.; Cui, C.; Lin, H.; Wu, Z.-L.* Enhancing the thermal stability of ketoreductase ChKRED12 using the FireProt web server. Process Biochemistry, 2021, 101:207-212

17.   Cui, C.; Lin, H. ; Pu, W. ;  Guo, C. ;  Liu, Y. ; Pei, X.-Q. ;  Wu, Z.-L.* Asymmetric Epoxidation and Sulfoxidation Catalyzed by a New Styrene Monooxygenase from Bradyrhizobium. Applied Biochemistry and Biotechnology, 2021, 193: 65-78.

18.   Li , C.; Pei, X.-Q.; Yi, D.; Li, T.-B.; Wu, Z.-L. Bioreductive dynamic kinetic resolution of ethyl 2-methoxy-3-oxo-3-phenylpropanoate. Catalysis Communications 135 (2020) 105865

19.   Cui, C.; Guo, C.; Lin, H.; Ding, Z.-Y.; Liu, Y.; Wu, Z.-L.* Functional characterization of an (R)-selective styrene monooxygenase from streptomyces sp. NRRL S-31. Enzyme and Microbial Technology, 2020, 132：109391

20.   Li, T.-B.; Zhao, F.-J.; Liu, Z.-C.; Jin, Y.; Liu, Y.; Pei, X.-Q.; Zhang, Z.-G.; Wang, G.-G*.; Wu, Z.-L.* Structure-guided engineering of ChKRED20 from Chryseobacterium sp. CA49 for asymmetric reduction of aryl ketoesters. Enzyme and Microbial Technology, 2019, 125: 29-36.

21.   Ren, Z.-Q.; Liu, Y.; Pei, X.-Q.; Wu, Z.-L.* Stereoselective bioreduction of ethyl 3-oxo-3-(2-thienyl) propanoate using the short-chain dehydrogenase/reductase ChKRED12. J. Microbiol. Biotechnol. (2019), 29(11), 1769–1776

22.   Pu, W.; Cui, C.; Guo, C.; Wu, Z.-L.* Characterization of two styrene monooxygenases from marine microbes. Enzyme and Microbial Technology, 2018, 112:29-34.

23.   Zhao, F.-J.; Jin, Y.; Liu, Z.-C.; Guo, C.; Li, T.-B.; Li, Z.-Y.; Wang G.-G.*; Wu, Z.-L*. Crystal structure and iterative saturation mutagenesis of ChKRED20 for expanded catalytic scope. Appl Microbiol Biotechnol. 2017, 101(23–24): 8395–8404.

24.   Guo, C.; Wu, Z.-L.* Construction and functional analysis of a whole-cell biocatalyst based on CYP108N7. Enzyme and Microbial Technology, 2017, 106：28-34.

25.   Liu, Y.-C.; Guo, C.; Liu, Y.; Wang, H.-B.; Wu, Z.-L.* Enzymatic cascades for the stereo-complementary epimerisation of in situ generated epoxy alcohols. Org. Biomol. Chem., 2017, 15, 2562-2568.

26.   Li, C.; Liu, Y.; Pei, X.-Q.; Wu, Z.-L.* Stereo-complementary bioreduction of saturated N-heterocyclic ketones, Process Biochem., 2017, 56：90-97.

27.   Zhao, F.-J.; Liu, Y.; Pei, X.-Q.;Guo, C.; Wu, Z.-L.* Single mutations of ketoreductase     ChKRED20 enhance the bioreductive production of (1S)-2-chloro-1-(3,4-difluorophenyl) ethanol. Appl Microbiol Biotechnol, 2017,101（5）：1945-1952.

28.   Liu, Y.; Liu, Y.-C; Wu, Z.-L*. Asymmetric bio‑epoxidation catalyzed with the styrene monooxygenase from Pseudomonas sp. LQ26. Bioresour. Bioprocess. 2016, 100（8）:3567-3575.

29.   Xu, G.-P.; Wang, H.-B., Wu, Z.-L*. Efficient bioreductive production of (S)-N-Boc-3-hydroxypiperidine using ketoreductase ChKRED03. Process Biochem. 2016, 51: 881-885.

30.   Pei, X.-Q.; Xu, M.-Y.; Wu, Z.-L.* Two “classical” Old Yellow Enzymes from Chryseobacterium sp. CA49: broad substrate specificity of Chr-OYE1 and limited activity of Chr-OYE2.  J. Mol. Catal. B: Enzym. 2016, 123:91-99. 

31.   Zhao, F.-J.; Pei, X.-Q.; Ren, Z.-Q.; Wu, Z.-L.* Rapid asymmetric reduction of ethyl 4-chloro-3-oxobutanoate using a thermostabilized mutant of ketoreductase ChKRED20. Appl Microbiol Biotechnol. 2016. 100 (8): 3567-3575.

32.   Liu, Y.-C; Wu, Z.-L*. Switchable asymmetric bio-epoxidation of ,-unsaturated ketones. Chem. Commun. 2016, 52(6), 1158 – 1161.

33.   Li, J.-J.; Pei, X.-Q.; Zhang, S.-B.; Wu, Z.-L*. Improving the thermostability of feruloyl esterase by DNA shuffling and site-directed mutagenesis. Process Biochem. 2015, 50: 1783-1787.

34.   Li, J.-J.; Yi, Z.-L.; Pei, X.-Q.; Wu, Z.-L*. Removal of the free cysteine residue reduces irreversible thermal inactivation of Feruloyl Esterase: evidence from circular dichroism and fluorescence spectra. Acta Biochimica et Biophysica Sinica. 2015, 47(8):612-619.

35.   Ren, Z.-Q; Liu, Y.; Pei, X.-Q; Wang H.-B.; Wu, Z.-L*. Bioreductive production of enantiopure (S)-duloxetine intermediatescatalyzed with ketoreductase ChKRED15. J. Mol. Catal. B: Enzym. 2015, 113,76–81.

36.   Liu, Y.-C; Liu, Y.; Wu, Z.-L*. Synthesis of enantiopure glycidol derivatives via a one-pot two-step enzymatic cascade.  Org. Biomol. Chem., 2015, 13, 2146–2152

37.   Xu, M.-Y., Pei, X.-Q., Wu, Z.-L.* Identification and characterization of a novel "thermophilic-like" Old Yellow Enzyme from the genome of Chryseobacterium sp. CA49. J. Mol. Catal. B: Enzym. 2014, 108, 64-71.

38.   Tang, T.-X.; Liu, Y; Wu, Z.-L.* Characterization of a robust anti-Prelog short-chain dehydrogenase/reductase ChKRED20 from Chryseobacterium sp. CA49. J. Mol. Catal. B: Enzym. 2014, 105, 82-88.

39.   Liu, Y., Tang, T.-X., Pei, X.-Q., Zhang, C., Wu, Z.-L.* Identification of ketone reductase ChKRED20 from the genome of Chryseobacterium sp. CA49 for highly efficient anti-Prelog reduction of 3,5-bis(trifluoromethyl)acetophenone. J. Mol. Catal. B: Enz. 2014, 102(4), 1-8.

40.   Wang, H.-B., Pei, X.-Q.; Wu, Z.-L.* An enoate reductase Acrh-OYE4 from Achromobacter sp. JA81: Characterization and application in asymmetric bioreduction of C=C bonds. Appl. Microbiol. Biotechnol. 2014, 98(2), 705-715.

41.   Yi, Z.-L.; Zhang, S.-B.*; Pei, X.-Q.; Wu, Z.-L.* Design of mutants for enhanced thermostability of b-glycosidase BglY from Thermus thermophilus. Bioresour. Technol. 2013, 129, 629-633.

42.   Lin, H.; Tang, D.-F.; Ahmed, A. A. Q.; Liu, Y.; Wu, Z.-L.* Mutations at the putative active cavity of styrene monooxygenase: enhanced activity and reversed enantioselectivity. J. Biotechnol. 2012, 161, 235-241.

43.   Zhang, S.-B.; Pei, X.-Q.; Wu, Z.-L.* Multiple amino acid substitutions significantly improve the thermostability of feruloyl esterase A from Aspergillus niger. Bioresour. Technol. 2012, 117, 140-147.

44.   Liu, Y.-J. （#）; Pei, X.-Q. （#）; Lin, H.; Gai, P.; Liu, Y.-C.; Wu, Z.-L.* Asymmetric bioreduction of activated alkenes by a novel isolate of Achromobacter species producing enoate reductase. Appl. Microbiol. Biotechnol. 2012, 95(3), 635-645.

45.   Zhang, S.-B.; Ma, X.-F.; Pei, X.-Q.; Liu, J.-Y.; Shao, H.-W.*, Wu, Z.-L.* A practical high-throughput screening system for feruloyl esterases: substrate design and evaluation. J. Mol. Catal. B: Enzym. 2012, 74, 36-40.

46.   Lin, H.; Liu, J.-Y.; Wang, H.-B.; Ahmed, A. A. Q.; Wu, Z.-L.* Biocatalysis as an alternative for the production of chiral epoxides: a comparative review. J. Mol. Catal. B: Enzym. 2011, 72(3-4), 77-89. (Invited review)

47.   Tang, C.-G.; Lin, H.; Zhang, C.; Liu, Z.-Q.; Yang, T.; Wu, Z.-L.* Highly enantioselective bioreduction of N-methyl-3-oxo-3- (thiophen-2-yl) propanamide for the production of (S)-duloxetine. Biotechnol. Lett. 2011, 33(7), 1435-1440.

48.   Lin, H.; Liu, Y.; Wu, Z.-L.* Highly diastereo- and enantio-selective epoxidation of secondary allylic alcohols catalyzed by styrene monooxygenase. Chem. Commun., 2011, 47, 2610-2612.

49.   Lin, H.; Liu, Y.; Wu, Z.-L.* Asymmetric epoxidation of styrene derivatives by styrene monooxygenase from Pseudomonas sp. LQ26: effects of - and -substituents. Tetrahedron: Asymmetry, 2011, 22, 134-137.

50.   Yi, Z.-L.; Pei, X.-Q.; Wu, Z.-L.* Introduction of glycine and proline residues onto protein surface increases the thermostability of endoglucanase CelA from Clostridium thermocellum. Bioresour. Technol. 2011, 102(3), 3636–3638

51.   Pei, X.-Q.; Yi, Z.-L.; Tang, C.-G.; Wu, Z.-L.* Three amino acid changes contribute markedly to the thermostability of -glucosidase BglC from Thermobifida fusca. Bioresour. Technol. 2011, 102(3), 3337–3342.

52.   Qaed, A. A.; Lin, H.; Tang, D.-F.; Wu, Z.-L.* Rational design of styrene monooxygenase mutants with altered substrate preference. Biotechnol. Lett. 2011, 33, 611-616.

53.   Zhang, S.-B.; Wu, Z.-L.* Identification of amino acid residues responsible for increased thermostability of feruloyl esterase A from Aspergillus niger using the PoPMuSiC algorithm.   Bioresour. Technol. 2011, 102 (2), 2093–2096.

54.   Lin, H.; Qiao, J.; Liu, Y.; Wu, Z.-L.* Styrene monooxygenase from Pseudomonas sp. LQ26 catalyzes the asymmetric epoxidation of both conjugated and unconjugated alkenes. J. Mol. Catal. B: Enzym. 2010, 67(3-4), 236-241.

55.   Zhang, Z.-G.; Yi, Z.-L.; Pei, X.-Q.; Wu, Z.-L.* Improving the thermostability of Geobacillus stearothermophilus xylanase XT6 by directed evolution and site-directed mutagenesis. Bioresour. Technol. 2010, 101(23), 9272-9278.

56.   Yi, Z.-L.; Wu, Z.-L.* Mutations from a family-shuffling-library reveal amino acid residues responsible for the thermostability of endoglucanase CelA from Clostridium thermocellum Biotechnol. Lett. 2010, 32(12), 1869-1875.

57.   Tang, Z., Salamanca-Pinzon, S.G., Wu, Z.L., Xiao, Y., Guengerich, F.P., Human cytochrome P450 4F11: Heterologous expression in bacteria, purification, and characterization of catalytic function. Arch. Biochem. Biophys. 2010, 494(1), 86-93.

58.   Wu, Z.-L.*; Qiao, J.; Zhang, Z.-G.; Guengerich, F.P.; Liu, Yan; Pei, X.-Q. Enhanced bacterial expression of several mammalian cytochrome P450s by codon optimization and chaperone coexpression. Biotechnol. Lett. 2009, 31(10), 1589-1593.

59.   Zhang, Z.-G.; Liu, Y.; Guengerich, F.P.; Matse, J.H.; Chen J.; Wu, Z.-L.* Identification of amino acid residues involved in 4-chloroindole 3-hydroxylation by cytochrome P450 2A6 using screening of random libraries. J. Biotechnol. 2009, 139(1), 12-18.

60.   Stark, K.; Wu, Z.-L.; Bartleson, C.J.; Guengerich, F.P. Heterologous expression and mRNA distribution of orphan cytochrome P450 20A1. Drug Metab. Dispos. 2008, 36(9), 1930-1937.

61.   Wu, Z.-L.; Sohl, C.D.; Shimada, T.; Guengerich, F.P. Recombinant Enzymes Over expressed in Bacteria Show Broad Catalytic Specificity of Human Cytochrome P450 2W1 and Limited Activity of Human Cytochrome P450 2S1. Mol. Pharmacol., 2006, 69(6), 2007-2014.

62.   Wu, Z.-L.; Bartleson, C.J.; Ham, A.J.; Guengerich, F.P. Heterologous Expression, Purification, and Properties of human cytochrome P450 27C1, Arch. Biochem. Biophys. 2006, 445(1), 138-146.

63.   Wu, Z.-L.; Podust, L.M.; Guengerich, F.P. Expansion of Substrate Specificity of Cytochrome P450 2A6 by Random and Site-Directed Mutagenesis. J. Biol. Chem. 2005, 280(49), 41090-41100.

64.   Kim, D.; Wu, Z.-L.; Guengerich, F.P. Analysis of coumarin 7-hydroxylation activity of cytochrome P450 2A6 using random mutagenesis. J. Biol. Chem. 2005, 280(48), 40319-40327.

65.   Wu, Z.-L.; Aryal, P.; Lozach, O.; Meijer, L.; Guengerich, F.P. Biosynthesis of New Indigoid Inhibitors of Protein Kinases Using Recombinant Cytochrome P450 2A6, Chem. Biodiv. 2005, 2(1), 51-65.

66.   Guengerich, F.P.; Wu, Z.-L.; Bartleson, C.J. Function of Human Cytochrome P450s: Characterization of the Orphans. Biochem. Biophys. Res. Commun. 2005, 338(1), 465-469.

67.   Wu, Z.-L.; Li, Z.-Y. Highly Enantioselective Synthesis of ,-Disubstituted Malonamic Acids through Asymmetric Hydrolysis of Dinitriles with Rhodococcus sp. CGMCC 0497, Chem. Commun. 2003, 3, 386-387. (Selected by ChemInform, 2003, 34(23), Nitriles Q0520.)

68.   Wu, Z.-L.; Li, Z.-Y. Practical Synthesis of Optically Active ,-Disubstituted Malonamic Acids through Asymmetric Hydrolysis of Malonamide Derivatives with Rhodococcus sp. CGMCC 0497, J. Org. Chem. 2003, 68(6), 2479-2482.

69.   Wu, Z.-L.; Li, Z.-Y. Biocatalytic Asymmetric Hydrolysis of ()--Hydroxy Nitriles by Rhodococcus sp. CGMCC 0497, J. Mol. Catal. B: Enzym. 2003, 22(1-2), 105-112.

70.   Wu, Z.-L.; Li, Z.-Y. Enantioselective Biotransformation of ,-Disubstituted Dinitriles to the Corresponding 2-Cyano-Acetamides using Rhodococcus sp. CGMCC 0497, Tetrahedron: Asymmetry, 2003, 14(15), 2133-2142.

71.   Wu, Z.-L.; Li, Z.-Y. Enhancement of Enzyme Activity and Enantioselectivity via Cultivation in Nitrile Metabolism by Rhodococcus sp. CGMCC 0497, Biotechnol. Appl. Biochem. 2002, 35(1), 61-67.

72.   Wu, Z.-L.; Li, Z.-Y. Enantioselective Hydrolysis of Various Racemic *-Substituted Arylacetonitriles using Rhodococcus sp. CGMCC 0497, Tetrahedron: Asymmetry 2001, 12(23), 3305-3312.

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