用于新兴污染物降解的酶纳米杂化物.pdf

1、Enzyme Nanohybrids for Emerging Contaminant DegradationYifan Gao,Meng Wang,Kshitija Shah,Shaily MahendraUCLA Civil and Environmental EngineeringAcknowledgmentsUCLA Faculty:Leonard Rome,Bruce Dunn,Eric Hoek,Patrick Allard,Chong Liu,JunyoungPark,Valerie KickhoeferUCLA Students and postdocs:Benjamin Cr

2、oze,Yichang Chen,Esther Lan,David Zgonc,Minhao Xiao,Haiyuan Zou,Glenn Nurwono,Yu Chen,Randy Chen,Jingyu Wang,Yongchao XieUSEPA:Quinn Birch,Mallikarjuna NadagoudaUniversity of Georgia:Qingguo HuangMicrobial Enzymes Biotransform Contaminants3Created by BioRNarayanan et al.,JEM,2023Enzyme Immobilizatio

3、n4Surface BindingCrosslinkingEntrapment+-GlassPolymersActivated CarbonNanoparticlesImmobilizationEnzymes immobilized into/ontonanomaterials for stability and longevityYifan Gao et al.,Biotech.Adv.,2022Nanoscale:Size Matters Nanoscale=1-100 nm in at least 1 dimension Increased surface area=increase r

4、eactivity Wave/Particle duality=unique nanoscale propertiesKlaine et al.2011The ratio of a nanomaterial to a tennis ball is about the same as the ratio of tennis ball to the moon!Human hair grows6 inches/year=5 nm/sec!Vault NanoparticlesFrascotti et al.,Cancers,2021;Kickhoefer et al.,PNAS,20056vPARP

5、INT serves as a“Zip code”to help deliver proteinsinto vaultsC-terminalN-terminalMajor vaultprotein(MVP)INTProtein of interestEngineered emptyvaults7Cooper sitesLaccaseCiprofloxacinProteins of Interest:Fungal Laccase and MnPLigninMnPMot and Silaghi-Dumitrescu,2012;Gecco BiotechLets make recombinant p

6、roteinsYifan Gao et al.,Bioresour.Technol.,20228Transfection of Insect CellsLaccaseLigationLaccase-INT-pFastBacPlasmidBacmidE.coliTransformationIsolationRecombinant Laccase-INT-BacmidLaccase-INTBaculovirusLaccaseINTRecombinantLaccase-INTInfected Sf9 CellsProduce recombinant laccase-INT in Sf9 cellsS