Optimization of process variables by central composite design for the immobilization of urease enzyme on functionalized gold nanoparticles for various applications
Tóm tắt
In the present study, enzyme urease has been immobilized on amine-functionalized gold nanoparticles (AuNPs). AuNPs were synthesized using natural precursor, i.e., clove extract and amine functionalized through 0.004 M l-cysteine. Enzyme (urease) was extracted and purified from the vegetable waste, i.e., seeds of pumpkin to apparent homogeneity (sp. activity 353 U/mg protein). FTIR spectroscopy and transmission electron microscopy was used to characterize the immobilized enzyme. The immobilized enzyme exhibited enhanced activity as compared with the enzyme in the solution, especially, at lower enzyme concentration. Based on the evaluation of activity assay of the immobilized enzyme, it was found that the immobilized enzyme was quite stable for about a month and could successfully be used even after eight cycles having enzyme activity of about 47%. In addition to this central composite design (CCD) with the help of MINITAB® version 15 Software was utilized to optimize the process variables viz., pH and temperature affecting the enzyme activity upon immobilization on AuNPs. The results predicted by the design were found in good agreement (R
2 = 96.38%) with the experimental results indicating the applicability of proposed model. The multiple regression analysis and ANOVA showed the individual and cumulative effect of pH and temperature on enzyme activity indicating that the activity increased with the increase of pH up to 7.5 and temperature 75 °C. The effects of each variables represented by main effect plot, 3D surface plot, isoresponse contour plot and optimized plot were helpful in predicting results by performing a limited set of experiments.
Tài liệu tham khảo
Daniel M, Astruc D (2004) Gold nanoparticles: assembly, supramolecular chemistry, quantum-size-related properties, and applications toward biology, catalysis, and nanotechnology. Chem Rev 104:293–346
Park SJ, Taton TA, Mirkin CA (2002) Scanometric DNA array detection with nanoparticle probe. Science 295:1757–1760
Huang GS, Chen YS, Yeh HW (2006) Measuring the flexibility of immunoglobulin by gold nanoparticles. Nano Lett 6:2467–2471
Gao XH, Cui YY, Levenson RM, Chung LWK, Nie SM (2004) In vivo cancer targeting and imaging with semiconductor quantum dots. Nat Biotechnol 2:969–976
Rosi NL, Giljohann DA, Thaxton CS, Lytton-Jean AKR, Han MS, Mirkin CA (2006) Oligonucliotide-modified gold nanoparticles for intracellular gene regulation. Science 312:1027–1030
Baron R, Willner B, Willner I (2007) Biomolecule-nanoparticle hybrids as functional units for nanobiotechnology. Chem Commun 17:323–332
Niemeyer CM (2001) Nanoparticles, proteins, and nucleic acids: biotechnology meets materials science. Angew Chem Int Ed 40:4128–4158
Selvakannan PR, Mandal S, Phadtare S, Gole A, Pasricha R, Sastry M (2004) Water-dispersible tryptophan-protected gold nanoparticles prepared by the spontaneous reduction of aqueous chloroaurate ions by the amino acid. J Colloid Interface Sci 269:97–102
Polavarapu L, Xu Q (2008) A single-step synthesis of gold nanochains using an amino acid as a capping agent and characterization of their optical properties. Nanotechnology 19:075601–075608
Zhou X, Yu T, Zhang Y, Kong J, Tang Y, Marty JL, Liu B (2007) Nanozeolite-assembled interface towards sensitive biosensing. Electrochem Commun 9:525–1529
Ciurli S, Marzadori C, Benini S, Deiana S, Gessa C (1996) Urease from the soil bacterium bacillus Pasteur II: immobilization on Ca-polygalacturonate. Soil Biol Biochem 28:811–817
Gay Bayrami L, Emine Y, Yakup Arica M (2005) Immobilization of urease via adsorption onto l-histidine–Ni(II) complexed poly(HEMA-MAH) microspheres: preparation and characterization. Process Biochem 23:3505–3513
Prakash O, Talat M, Hasan SH, Pandey RK (2008) Enzymatic detection of mercuric ions in ground-water from vegetable wastes by immobilizing pumpkin (Cucumis melo) urease in calcium alginate beads. Bioresour Technol 99:4524–4528
Prakash O, Talat M, Hasan SH (2008) Response surface design for the optimization of enzymatic detection of mercury ions in aqueous solution using immobilized Urease from vegetable waste. J Mol Catal B Enzym 56:265–271
Ranjan D, Srivastava P, Talat M, Hasan SH (2009) Removal of Cr(VI) from water using biomass of aeromonas hydrophila: central composite design for optimization of process variables. Appl Biochem Biotechnol 158:524–539
Ranjan D, Srivastava P, Talat M, Hasan SH (2009) Biosorption of Arsenic from aqueous solution using agricultural residue “Rice Polish”. J Hazard Mater 166:1050–1059
Talat M, Prakash O, Hasan SH (2009) Enzymatic detection of As(III) in aqueous solution using alginate immobilized pumpkin urease: optimization of process variables by response surface methodology. Bioresour Technol 100:4462–4467
Singh AK, Talat M, Singh DP, Srivastava ON (2010) Biosynthesis of silver and gold nanoparticles by novel natural precursor Clove. J Nanopart Res 12:1667–1675
Prakash O, Upadhyay LSB (2005) Physico-chemical characterization of watermelon urease upon immobilization in alginate beads. J Plant Biochem Biotechnol 14:209–213
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275
Sheng J, Chi Z, Yan K, Wang X, Gong F, Li J (2009) Use of response surface methodology for optimizing process parameters for high inulinase production by the marine yeast Cryptococcus aureus G7a in solid-state fermentation and hydrolysis of inulin. Bioprocess Biosyst Eng 32:333–339
Zhu Y, Ni N, Huang W (2010) Process optimization for the production of diosgenin with Trichoderma reesei. Bioprocess Biosyst Eng 33:647–655
Du D, Liu S, Chen Ji, Ju H, Lian H, Li J (2005) Colloidal gold nanoparticle modified carbon paste interface for studies of tumor cell adhesion and viability. Biomaterials 26:6487–6495
Sastry M, Ahmad A, Khan MI, Kumar R (2003) Biosynthesis of metal nanoparticles using fungi and actinomycete. Curr Sci 85:162–170
Gole A, Dash C, Ramachandran V, Mandale AB, Sainkar SR, Rao M, Sastry M (2001) Pepsin–gold colloid conjugates: preparation, characterization and enzyme activity. Langmuir 17:1674–1679
Vigneshwaran N, Kathe AA, Varadarajan PV, Nachane RP, Balasubramanya RH (2007) Silver-protein (core-shell) nanoparticle production using spent mushroom substrate. Langmuir 23:7113–7117
Shankar SS, Ahmad A, Sastry M (2003) Geranium leaf assisted biosynthesis of silver nanoparticles. Biotechnol Prog 19:1627–1631
JingJing Z, JunJie Z (2009) A novel amperometric biosensor based on gold nanoparticles–mesoporous silica composite for biosensing glucose. Sci China Ser B-Chem 52:815–820
Huang SH, Liao MH, Chen DH (2003) Direct binding and characterization of lipase onto magnetic nanoparticles. Biotechnol Prog 3:1095–1100
Guo Z, Bai S, Sun Y (2003) Preparation and characterization of immobilized lipase on magnetic hydrophobic microspheres. Enzyme Microb Technol 32:776–782
Leaka J, Wlodarczyk J, Zaborska W (1999) Polyaniline as a support for urease immobilization. J Mol Catal B Enzym 16:549–556
Zulkali MMD, Ahmad AL, Norulakmal NH (2006) Oryza sativa L. husk as heavy metal adsorbent: optimization with lead as model solution. Bioresour Technol 97:21–25
Jinhui S, Xie H, Wu W, Joseph VR, Jeff CF, Wang ZL (2010) Robust optimization of the output voltage of nanogenerators by statistical design of experiments. Nano Res 3(9):613–619
Ravikumar K, Krishnan S, Ramalingam S, Balu K (2007) Optimization of process variables by the application of response surface methodology for dye removal using a novel adsorbent. Dyes Pigm 72(1):66–70