Abstract
In this study, hydrophobic macroporous resin NKA was employed as matrix for immobilization of free Rhizopus oryzae lipase (ROL). The performance of the immobilized ROL was significantly enhanced. The recovery activity was up to 1,293.78 % and the specific activity increased to 152,914 U/g-protein, which was 46-fold higher than that of the free lipase. Moreover, the immobilized lipase showed higher thermostability and better pH-resistance than its free counterpart. Additionally, three different nonaqueous modification strategies (including bioimprinting, lecithin coating, and lyophilization protection) were further utilized to improve the performance of the immobilized lipase. The corresponding enhancements were 33.68 %, 31.98 %, and 99.86 %. When these modifications were combined together, the activity improved 209.51 %. In order to confirm its practical application, the modified ROL was used to biorefine rapeseed oil deodorizer distillate (RODD) for biodiesel production. The highest conversion yield reached 98.23 %, much close to that (97.46 %) of Novozym 435. The results suggest that the prepared lipase in this study is a promising biocatalyst with high stability, efficiency and operational reusability.
Similar content being viewed by others
Abbreviations
- DD:
-
Deodorizer distillate
- RODD:
-
Rapeseed oil deodorizer distillate
- FFA:
-
Free fatty acid
- ROL:
-
Rhizopus oryzae lipase
- Mtpa:
-
Million tons per annum
- BSA:
-
Bovine serum albumin
- NOV 435:
-
Novozym 435
- FAEEs:
-
Fatty acid ethyl esters
References
Li J, Li L, Tong J, Wang Y, Chen S (2012) Research development on lipase-catalyzed biodiesel. Energy Procedia 16, Part B: 1014–1021
Tecelão C, Guillén M, Valero F, Ferreira-Dias S (2012) Immobilized heterologous Rhizopus oryzae lipase: A feasible biocatalyst for the production of human milk fat substitutes. Biochem Eng J 67:104–110
Nunes PA, Pires-Cabral P, Guillén M, Valero F, Ferreira-Dias S (2012) Batch operational stability of immobilized heterologous Rhizopus oryzae lipase during acidolysis of virgin olive oil with medium-chain fatty acids. Biochem Eng J 67:265–268
Hama S, Tamalampudi S, Fukumizu T, Miura K, Yamaji H, Kondo A, Fukuda H (2006) Lipase localization in Rhizopus oryzae cells immobilized within biomass support particles for use as whole-cell biocatalysts in biodiesel-fuel production. J Biosci Bioeng 101:328–333
Fukuda H, Hama S, Tamalampudi S, Noda H (2008) Whole-cell biocatalysts for biodiesel fuel production. Trends Biotechnol 26:668–673
Kaieda M, Samukawa T, Matsumoto T, Ban K, Kondo A, Shimada Y, Noda H, Nomoto F, Ohtsuka K, Izumoto E, Fukuda H (1999) Biodiesel fuel production from plant oil catalyzed by Rhizopus oryzae lipase in a water-containing system without an organic solvent. J Biosci Bioeng 88:627–631
Bajaj A, Lohan P, Jha PN, Mehrotra R (2010) Biodiesel production through lipase catalyzed transesterification: An overview. J Mol Catal B-Enzym 62:9–14
Fjerbaek L, Christensen KV, Norddahl B (2009) A review of the current state of biodiesel production using enzymatic transesterification. Biotechnol Bioeng 102:1298–1315
Tan T, Lu J, Nie K, Deng L, Wang F (2010) Biodiesel production with immobilized lipase: A review. Biotechnol Adv 28:628–634
Jegannathan KR, Abang S, Poncelet D, Chan ES, Ravindra P (2008) Production of Biodiesel Using Immobilized Lipase—A Critical Review. Crit Rev Biotechnol 28:253–264
Bornscheuer UT, Bessler C, Srinivas R, Hari Krishna S (2002) Optimizing lipases and related enzymes for efficient application. Trends Biotechnol 20:433–437
Fernandez-Lorente G, Cabrera Z, Godoy C, Fernandez-Lafuente R, Palomo JM, Guisan JM (2008) Interfacially activated lipases against hydrophobic supports: effect of the support nature on the biocatalytic properties. Process Biochem 43:1061–1067
Sharma R, Chisti Y, Banerjee UC (2001) Production, purification, characterization, and applications of lipases. Biotechnol Adv 19:627–662
Yan Y, Zhang X, Chen D (2013) Enhanced catalysis of Yarrowia lipolytica lipase LIP2 immobilized on macroporous resin and its application in enrichment of polyunsaturated fatty acids. Bioresource Technol 131:179–187
Liu Y, Liu T, Wang X, Xu L, Yan Y (2011) Biodiesel synthesis catalyzed by Burkholderia cenocepacia lipase supported on macroporous resin NKA in solvent-free and isooctane systems. Energy Fuel 25:1206–1212
Wu JC, Lee SS, Mahmood MMB, Chow Y, Talukder MMR, Choi WJ (2007) Enhanced activity and stability of immobilized lipases by treatment with polar solvents prior to lyophilization. J Mol Catal B-Enzym 45:108–112
Okahata Y, Mori T (1997) Lipid-coated enzymes as efficient catalysts in organic media. Trends Biotechnol 15:50–54
Foresti ML, Alimenti GA, Ferreira ML (2005) Interfacial activation and bioimprinting of Candida rugosa lipase immobilized on polypropylene: effect on the enzymatic activity in solvent-free ethyl oleate synthesis. Enzyme Microb Tech 36:338–349
Gupta MN, Roy I (2004) Enzymes in organic media. Eur J Biochem 271:2575–2583
Yilmaz E (2002) Bio-imprinting of microbial lipase at air–water interface. World J Microb Biot 18:141–145
Pagani MA, Baltanás MA (2010) Production of natural antioxidants from vegetable oil deodorizer distillates: Effect of catalytic hydrogenation. Bioresource Technol 101:1369–1376
Kasim NS, Gunawan S, Ju Y (2009) Isolation and identification of steroidal hydrocarbons in soybean oil deodorizer distillate. Food Chem 117:15–19
Shao P, Jiang ST, Ying YJ (2007) Optimization of molecular distillation for recovery of tocopherol from rapeseed oil deodorizer distillate using response surface and artificial neural network models. Food Bioprod Process 85:85–92
Jiang ST, Shao P, Pan LJ, Zhao YY (2006) Molecular distillation for recovering tocopherol and fatty acid methyl esters from rapeseed oil deodoriser distillate. Biosyst Eng 93:383–391
Wang L, Du W, Liu D, Li L, Dai N (2006) Lipase-catalyzed biodiesel production from soybean oil deodorizer distillate with absorbent present in tert-butanol system. J Mol Catal B-Enzym 43:29–32
Facioli NL, Barrera-Arellano D (2001) Optimisation of enzymatic esterification of soybean oil deodoriser distillate. J Sci Food Agr 81:1193–1198
Ramamurthi S, McCurdy A (1993) Enzymatic pretreatment of deodorizer distillate for concentration of sterols and tocopherols. J Am Oil Chem Soc 70:287–295
Petkar M, Lali A, Caimi P, Daminati M (2006) Immobilization of lipases for non-aqueous synthesis. J Mol Catal B-Enzym 39:83–90
Liu T, Liu Y, Wang X, Li Q, Wang J, Yan Y (2011) Improving catalytic performance of Burkholderia cepacia lipase immobilized on macroporous resin NKA. J Mol Catal B-Enzym 71:45–50
Salis A, Svensson I, Monduzzi M, Solinas V, Adlercreutz P (2003) The atypical lipase B from Candida antarctica is better adapted for organic media than the typical lipase from Thermomyces lanuginosa. Bba-Proteins Proteom 1646:145–151
Gomes FM, Pereira EB, de Castro HF (2003) Immobilization of lipase on chitin and its use in nonconventional biocatalysis. Biomacromolecules 5:17–23
Cheirsilp B, H-Kittikun A, Limkatanyu S (2008) Impact of transesterification mechanisms on the kinetic modeling of biodiesel production by immobilized lipase. Biochem Eng J 42:261–269
Yu H, Wu J, Ching C (2004) Enhanced activity and enantioselectivity of Candida rugosa lipase immobilized on macroporous adsorptive resins for ibuprofen resolution. Biotechnol Lett 26:629–633
Wang H, Wu H, Ho C, Weng X (2006) Cocoa butter equivalent from enzymatic interesterification of tea seed oil and fatty acid methyl esters. Food Chem 97:661–665
Bastida A, Sabuquillo P, Armisen P, Fernández-Lafuente R, Huguet J, Guisán JM (1998) A single step purification, immobilization, and hyperactivation of lipases via interfacial adsorption on strongly hydrophobic supports. Biotechnol Bioeng 58:486–493
Serdakowski AL, Dordick JS (2008) Enzyme activation for organic solvents made easy. Trends Biotechnol 26:48–54
Burke PA, Griffin RG, Klibanov AM (1992) Solid-state NMR assessment of enzyme active center structure under non-aqueous conditions. J Biol Chem 267:20057–20064
Triantafyllou AÖ, Wehtje E, Adlercreutz P, Mattiasson B (1995) Effects of sorbitol addition on the action of free and immobilized hydrolytic enzymes in organic media. Biotechnol Bioeng 45:406–414
Khmelnitsky YL, Welch SH, Clark DS, Dordick JS (1994) Salts dramatically enhance activity of enzymes suspended in organic solvents. J Am Chem Soc 116:2647–2648
Bedell BA, Mozhaev VV, Clark DS, Dordick JS (1998) Testing for diffusion limitations in salt-activated enzyme catalysts operating in organic solvents. Biotechnol Bioeng 58:654–657
Fishman A, Cogan U (2003) Bio-imprinting of lipases with fatty acids. J Mol Catal B-Enzym 22:193–202
Yu D, Tian L, Wu H, Wang S, Wang Y, Ma DFang X (2010) Ultrasonic irradiation with vibration for biodiesel production from soybean oil by Novozym 435. Process Biochem 45:519–525
Modi MK, Reddy JRC, Rao BVSKPrasad RBN (2007) Lipase-mediated conversion of vegetable oils into biodiesel using ethyl acetate as acyl acceptor. Bioresource Technol 98:1260–1264
Acknowledgments
The authors acknowledge the financial support of the National Natural Science Foundation of P. R. China (NSFC; Nos. 31070089, 31170078, and J1103514), the National High Technology Research and Development Program of P. R. China (863 Program; Nos. 2011AA02A204, 2013AA065805), and the Innovation Foundation of Shenzhen Government (JCYJ20120831111657864). Many thanks are indebted to Ms Chen Hong, from the Centre of Analysis and Test, Huazhong University of Science and Technology for biodiesel analysis.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Su, F., Li, G., Zhang, H. et al. Enhanced Performance of Rhizopus oryzae Lipase Immobilized on Hydrophobic Carriers and Its Application in Biorefinery of Rapeseed Oil Deodorizer Distillate. Bioenerg. Res. 7, 935–945 (2014). https://doi.org/10.1007/s12155-014-9415-y
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12155-014-9415-y