Immobilized enzymatic alcohol oxidation as a versatile reaction module for multienzyme cascades

Abstract

Enzymatic alcohol oxidation (EAO) is highly attractive thanks to its efficiency, selectivity, and sustainability benefits, but it is often neglected as a catalytic tool for practical production due to the instability and non-reusability of enzymes. Herein, a non-enantioselective alcohol dehydrogenase engineered from Candida parapsilosis (CpsADH) and a laccase from Trametes versicolor was immobilized on mesoporous silica nanoflowers (MSNs), fabricating CpsADH@MSNs (41 U/g_support) and laccase@MSNs (67 U/g_support) for EAO, respectively. The structural and functional properties of the MSNs endowed the immobilized enzymes with higher stability than free enzymes, and the relative activity of the immobilized enzyme was 52% and 63%, respectively, after being reused five times. The immobilized enzymes exhibited high activity, selectivity, and complementary substrate specificity in alcohol oxidation. The optimized EAO, as a versatile cascade module, was coupled with several other enzymatic transformations for multi-enzymatic synthesis of high value-added chemicals. The chiral alcohols and amines were produced with 99% ee and 84% to 98% ee, respectively, and (R)-benzoin and 2-furoic acid were prepared with 91% yield, 99% ee and 86% yield, respectively, demonstrating the synthetic utility of the immobilized enzymes.

Keywords

Alcohol oxidation / asymmetric synthesis / enzyme immobilization / multienzyme cascades / mesoporous silica nanoflowers

References

[1] Asthana M,Kumar A.Direct oxidation of alcohols catalysed by heterometallic complex [CuNi(bz)₃(bpy)₂] ClO₄ to aldehydes and ketones mediated by hydrogen peroxide as a terminal oxidant.Inorganica Chimica Acta2020;502:119286.

[2] Shaabani A,Afaridoun H.Highly selective aerobic oxidation of alkyl arenes and alcohols: cobalt supported on natural hydroxyapatite nanocrystals.RSC Adv2016;6:48396-404

[3] Mallat T.Oxidation of alcohols with molecular oxygen on solid catalysts.Chem Rev2004;104:3037-58

[4] Nakagawa K,Nakata T.Oxidation with Nickel Peroxide. I. oxidation of alcohols.J Org Chem1962;27:1597-601

[5] Lou JD.Selective oxidation of primary alcohols with chromium trioxide under solvent free conditions.Tetrahedron Lett2002;43:6095-7

[6] Uyanik M.Hypervalent iodine-mediated oxidation of alcohols.Chem Comm2009;2086-99

[7] Taylor RJ,Foot J.Tandem oxidation processes using manganese dioxide: discovery, applications, and current studies.Acc Chem Res2005;38:851-69.

[8] Hasanpour B,Feizpour F.Copper(II)-ethanolamine triazine complex on chitosan-functionalized nanomaghemite for catalytic aerobic oxidation of benzylic alcohols.Catal Lett2021;151:45-55

[9] Martı́n SE.Catalytic aerobic oxidation of alcohols by Fe(NO₃)₃-FeBr₃.Tetrahedron Lett2002;43:4475-9

[10] Qiu S,Xu H.Efficient catalytic oxidation of benzyl alcohol by tetrasubstituted cobalt phthalocyanine-MWCNTs composites.Solid State Sci2022;129:106905

[11] Schultz MJ,Jensen DR.Development and comparison of the substrate scope of Pd-catalysts for the aerobic oxidation of alcohols.J Org Chem2005;70:3343-52 PMCID:PMC2692733

[12] Personick ML,Biener MM,Madix RJ.Ozone-activated nanoporous gold: a stable and storable material for catalytic oxidation.ACS Catal2015;5:4237-41

[13] Parmeggiani C.Transition metal based catalysts in the aerobic oxidation of alcohols.Green Chem2012;14:547-64

[14] Polshettiwar V.Green chemistry by nano-catalysis.Green Chem2010;12:743-54

[15] Shibuya M,Suzuki I.2-azaadamantane N-oxyl (AZADO) and 1-Me-AZADO: highly efficient organocatalysts for oxidation of alcohols.J Am Chem Soc2006;128:8412-3

[16] Punniyamurthy T,Iqbal J.Recent advances in transition metal catalyzed oxidation of organic substrates with molecular oxygen.Chem Rev2005;105:2329-63.

[17] Liu J,Li Z.Recent advances in enzymatic oxidation of alcohols.Curr Opin Chem Biol2018;43:77-86

[18] Moa S.Quantum chemical study of mechanism and stereoselectivity of secondary alcohol dehydrogenase.J Inorg Biochem2017;175:259-66

[19] Kroutil W,Edegger K.Biocatalytic oxidation of primary and secondary alcohols.Adv Synth Catal2004;346:125-42

[20] Tian K.A simple biosystem for the high-yielding cascade conversion of racemic alcohols to enantiopure amines.Angew Chem Int Ed Engl2020;59:21745-51.

[21] Albarrán-velo J,Lavandera I.One-pot two-step chemoenzymatic deracemization of allylic alcohols using laccases and alcohol dehydrogenases.Mol Catal2020;493:111087

[22] González-granda S,Lavandera I.Laccase-mediated oxidations of propargylic alcohols. Application in the deracemization of 1-arylprop-2-yn-1-ols in combination with alcohol dehydrogenases.ChemCatChem2020;12:520-7

[23] Martínez-montero L,Gotor-fernández V.Stereoselective amination of racemic sec-alcohols through sequential application of laccases and transaminases.Green Chem2017;19:474-80

[24] Méndez-sánchez D,Lavandera I,Gotor-fernández V.Chemoenzymatic deracemization of secondary alcohols by using a TEMPO-Iodine-alcohol dehydrogenase system.ChemCatChem2015;7:4016-20.

[25] de Gonzalo G.Recent trends in synthetic enzymatic cascades promoted by alcohol dehydrogenases.Curr Opin Green Sust2021;32:100548.

[26] Schoemaker HE,Wubbolts MG.Dispelling the myths--biocatalysis in industrial synthesis.Science2003;299:1694-7

[27] Hartmann M.Immobilization of enzymes on porous silicas--benefits and challenges.Chem Soc Rev2013;42:6277-89

[28] Gao J,Zhou L.Monodisperse core-shell magnetic organosilica nanoflowers with radial wrinkle for lipase immobilization.Chem Eng J2017;309:70-9.

[29] Deng J,Zhan H.Catalyzed degradation of polycyclic aromatic hydrocarbons by recoverable magnetic chitosan immobilized laccase from Trametes versicolor.Chemosphere2022;301:134753

[30] Zheng F,Wu XJ,Liu HX.Immobilization of laccase onto chitosan beads to enhance its capability to degrade synthetic dyes.Int Biodeter Biodegr2016;10:69-78

[31] Gao J,Du Y.Construction of biocatalytic colloidosome using lipase-containing dendritic mesoporous silica nanospheres for enhanced enzyme catalysis.Chem Eng J2017;317:175-86

[32] Videira-quintela D,Montalvo G.Emerging opportunities of silica-based materials within the food industry.Microchem J2021;167:106318.

[33] Costantini A.Lipase immobilization in mesoporous silica nanoparticles for biofuel production.Catalysts2021;11:629

[34] Kalantari M,Yang Y.Tailoring mesoporous-silica nanoparticles for robust immobilization of lipase and biocatalysis.Nano Res2017;10:605-17.

[35] Cao G,Zhou L.Fabrication of Ni²⁺-nitrilotriacetic acid functionalized magnetic mesoporous silica nanoflowers for one pot purification and immobilization of His-tagged ω-transaminase.BioChem Eng J2017;128:116-25.

[36] Zhou L,Kong W.One pot purification and co-immobilization of His-tagged old yellow enzyme and glucose dehydrogenase for asymmetric hydrogenation.Enzyme Microb Technol2022;156:110001

[37] Li Y,Dong L.Asymmetric reduction of conjugated C=C bonds by immobilized fusion of old yellow enzyme and glucose dehydrogenase.Green Synth Catal2022;online ahead of print

[38] Gao L,Liu Y.Co-immobilization of metal and enzyme into hydrophobic nanopores for highly improved chemoenzymatic asymmetric synthesis.Chem Commun2020;56:13547-50

[39] Kong W,Huang C.Direct asymmetric reductive amination of alkyl (hetero)aryl ketones by an engineered amine dehydrogenase.Angewandte Chemie2022;134:e202202264

[40] Demir A,Eren E.Enantioselective synthesis of α-Hydroxy ketones via benzaldehyde lyase-catalyzed C−C bond formation reaction.Adv Synth Catal2002;344:96-103

[41] Peng B,Zhang PQ.An effective hybrid strategy for converting rice straw to furoic acid by tandem catalysis via Sn-sepiolite combined with recombinant E. coli whole cells harboring horse liver alcohol dehydrogenase.Green Chem2019;21:5914-23.

[42] Ghannadi S,Miroliaei M.Immobilization of alcohol dehydrogenase on titania nanoparticles to enhance enzyme stability and remove substrate inhibition in the reaction of formaldehyde to methanol.Ind Eng Chem Res2019;58:9844-54

[43] Esmaeilnejad-ahranjani P,Singh G.Amine-functionalized magnetic nanocomposite particles for efficient immobilization of lipase: effects of functional molecule size on properties of the immobilized lipase.RSC Adv2015;5:33313-27

[44] Zang L,Wu X,Sakai E.Preparation of magnetic chitosan nanoparticles as support for cellulase immobilization.Ind Eng Chem Res2014;53:3448-54

[45] Barbosa O,Berenguer-murcia Á,Rodrigues RC.Glutaraldehyde in bio-catalysts design: a useful crosslinker and a versatile tool in enzyme immobilization.RSC Adv2014;4:1583-600

[46] Songulashvili G,Jaspers C.Immobilized laccase of Cerrena unicolor for elimination of endocrine disruptor micropollutants.Fungal Biol2012;116:883-9

[47] Zhang J,Chen H.Removal of 2,4-dichlorophenol by chitosan-immobilized laccase from Coriolus versicolor.BioChem Eng J2009;45:54-9.

[48] Sathishkumar P,Cho M.Laccase immobilization on cellulose nanofiber: the catalytic efficiency and recyclic application for simulated dye effluent treatment.J Mol Catal B-Enzym2014;100:111-20.

[49] Kašpar O,Nyanhongo GS,Štěpánek F.Effect of cross-linking method on the activity of spray-dried chitosan microparticles with immobilized laccase.Food Bioprod Process2013;91:525-33

[50] Li X,Feng J.One-pot polylol synthesis of graphene decorated with size- and density-tunable Fe₃O₄ nanoparticles for porcine pancreatic lipase immobilization.Carbon2013;60:488-97

[51] Ida J,Yamamoto H.Immobilization of glucoamylase on ceramic membrane surfaces modified with a new method of treatment utilizing SPCP-CVD.Biochem Eng J2000;5:179-84

[52] Grahame DAS,Yada RY. Improving and tailoring enzymes for food quality and functionality. Woodhead Publishing; 2015.pp.11-55.

[53] Frankenberger W.Effect of pH on enzyme stability in soils.Soil Biol Biochem1982;14:433-7

[54] Puetz H,Vranková K.Biocatalytic oxidation of alcohols.Catalysts2020;10:952

[55] Lalonde J.Highly engineered biocatalysts for efficient small molecule pharmaceutical synthesis.Curr Opin Biotechnol2016;42:152-8

[56] Paul CE,Gotor‐Fernández V,Gotor V.Escherichia coli /ADH-A: an all-inclusive catalyst for the selective biooxidation and deracemisation of secondary alcohols.ChemCatChem2013;5:3875-81

[57] Musa MM,Mutti FG.Synthesis of enantiomerically pure alcohols and amines via biocatalytic deracemisation methods.Catal Sci Technol2019;9:5487-503. PMCID:PMC7116805

[58] Mutti FG,Scrutton NS,Turner NJ.Conversion of alcohols to enantiopure amines through dual-enzyme hydrogen-borrowing cascades.Science2015;349:1525-9 PMCID:PMC4883652

[59] Chen FF,Zheng GW.Asymmetric amination of secondary alcohols by using a redox-neutral two-enzyme cascade.ChemCatChem2015;7:3838-41.