Expression and characterization of a CALB-type lipase from Sporisorium reilianum SRZ2 and its potential in short-chain flavor ester synthesis
Jiang-Wei Shen, Xue Cai, Bao-Juan Dou, Feng-Yu Qi, Xiao-Jian Zhang, Zhi-Qiang Liu, Yu-Guo Zheng
Expression and characterization of a CALB-type lipase from Sporisorium reilianum SRZ2 and its potential in short-chain flavor ester synthesis
A lipase from Sporisorium reilianum SRZ2 (SRL) with 73% amino acid sequence identity to Candida antarctica lipase B (CALB) was cloned and overexpressed in Pichia pastoris. The recombinant SRL showed a preference for short-chain p-nitrophenyl esters. It achieved maximum activity at pH 8.0 and 65°C for p-nitrophenyl hexanoate (C6) with Km and kcat/Km values of 0.14 mmol∙L−1 and 1712 min−1∙mmol∙L−1 at 30°C, respectively. SRL displayed excellent thermostability and pH stability, retaining more than 79% of its initial activity after incubation at 60°C for 72 h and 75% at pH 3 to 11 for 72 h. It also maintained most of its activity in the presence of inhibitors and detergents except sodium dodecyl sulfate, and it tolerated organic solvents. SRL was covalently immobilized and successfully used for ethyl hexanoate synthesis in cyclohexane or in a solvent-free system with a high conversion yield (>95%). Furthermore, high conversion yield was also achieved for the synthesis of various short-chain flavor esters when high substrate concentrations of 2 mol∙L−1 were applied. This study indicated that a CALB-type lipase from S. reilianum SRZ2 showed great potential in organic ester synthesis.
lipase / Sporisorium reilianum / biochemical characterization / short-chain flavor ester / solvent-free system
[1] |
Adlercreutz P. Immobilisation and application of lipases in organic media. Chemical Society Reviews, 2013, 42(15): 6406–6436
CrossRef
Google scholar
|
[2] |
Stergiou P Y, Foukis A, Filippou M, Koukouritaki M, Parapouli M, Theodorou L G, Hatziloukas E, Afendra A, Pandey A, Papamichael E M. Advances in lipase-catalyzed esterification reactions. Biotechnology Advances, 2013, 31(8): 1846–1859
CrossRef
Google scholar
|
[3] |
Tan T, Lu J, Nie K, Deng L, Wang F. Biodiesel production with immobilized lipase: A review. Biotechnology Advances, 2010, 28(5): 628–634
CrossRef
Google scholar
|
[4] |
Fernández-Lafuente R. Lipase from Thermomyces lanuginosus: Uses and prospects as an industrial biocatalyst. Journal of Molecular Catalysis. B, Enzymatic, 2010, 62(3-4): 197–212
CrossRef
Google scholar
|
[5] |
Javed S, Azeem F, Hussain S, Rasul I, Siddique M H, Riaz M, Afzal M, Kouser A, Nadeem H. Bacterial lipases: A review on purification and characterization. Progress in Biophysics and Molecular Biology, 2018, 132: 23–34
CrossRef
Google scholar
|
[6] |
Anderson E M, Larsson K M, Kirk O. One biocatalyst—many applications: The use of Candida antarctica B-lipase in organic synthesis. Biocatalysis and Biotransformation, 1998, 16(3): 181–204
CrossRef
Google scholar
|
[7] |
Buerth C, Kovacic F, Stock J, Terfruchte M, Wilhelm S, Jaeger K E, Feldbrugge M, Schipper K, Ernst J F, Tielker D. Uml2 is a novel CalB-type lipase of Ustilago maydis with phospholipase A activity. Applied Microbiology and Biotechnology, 2014, 98(11): 4963–4973
CrossRef
Google scholar
|
[8] |
Vaquero M E, de Eugenio L I, Martinez M J, Barriuso J. A novel calb-type lipase discovered by fungal genomes mining. PLoS One, 2015, 10(4): e0124882
CrossRef
Google scholar
|
[9] |
Park S. Exploration and functional expression of homologous lipases of Candida antarctica lipase B. Korean Journal of Microbiology, 2015, 51(3): 187–193
CrossRef
Google scholar
|
[10] |
Dhake K P, Thakare D D, Bhanage B M. Lipase: A potential biocatalyst for the synthesis of valuable flavour and fragrance ester compounds. Flavour and Fragrance Journal, 2013, 28(2): 71–83
CrossRef
Google scholar
|
[11] |
Sá A G A, Meneses A C, Araújo P H H, Oliveira D. A review on enzymatic synthesis of aromatic esters used as flavor ingredients for food, cosmetics and pharmaceuticals industries. Trends in Food Science & Technology, 2017, 69: 95–105
CrossRef
Google scholar
|
[12] |
Serra S, Fuganti C, Brenna E. Biocatalytic preparation of natural flavours and fragrances. Trends in Biotechnology, 2005, 23(4): 193–198
CrossRef
Google scholar
|
[13] |
Gao W, Wu K, Chen L, Fan H, Zhao Z, Gao B, Wang H, Wei D. A novel esterase from a marine mud metagenomic library for biocatalytic synthesis of short-chain flavor esters. Microbial Cell Factories, 2016, 15(1): 41
CrossRef
Google scholar
|
[14] |
Yan H D, Zhang Q, Wang Z. Biocatalytic synthesis of short-chain flavor esters with high substrate loading by a whole-cell lipase from Aspergillus oryzae. Catalysis Communications, 2014, 45: 59–62
CrossRef
Google scholar
|
[15] |
Lozano P, Bernal J M, Navarro A. A clean enzymatic process for producing flavour esters by direct esterification in switchable ionic liquid/solid phases. Green Chemistry, 2012, 14(11): 3026–3033
CrossRef
Google scholar
|
[16] |
Martins A B, Friedrich J L, Cavalheiro J C, Garcia-Galan C, Barbosa O, Ayub M A, Fernández-Lafuente R, Rodrigues R C. Improved production of butyl butyrate with lipase from Thermomyces lanuginosus immobilized on styrene-divinylbenzene beads. Bioresource Technology, 2013, 134: 417–422
CrossRef
Google scholar
|
[17] |
Zheng Y G, Yin H H, Yu D F, Chen X, Tang X L, Zhang X J, Xue Y P, Wang Y J, Liu Z Q. Recent advances in biotechnological applications of alcohol dehydrogenases. Applied Microbiology and Biotechnology, 2017, 101(3): 987–1001
CrossRef
Google scholar
|
[18] |
Liu Z Q, Wu L, Zheng L, Wang W Z, Zhang X J, Jin L Q, Zheng Y G. Biosynthesis of tert-butyl (3R,5S)-6-chloro-3,5-dihydroxyhexanoate by carbonyl reductase from Rhodosporidium toruloides in mono and biphasic media. Bioresource Technology, 2018, 249: 161–167
CrossRef
Google scholar
|
[19] |
Liu Z Q, Lu M M, Zhang X H, Cheng F, Xu J M, Xue Y P, Jin L Q, Wang Y S, Zheng Y G. Significant improvement of the nitrilase activity by semi-rational protein engineering and its application in the production of iminodiacetic acid. International Journal of Biological Macromolecules, 2018, 116: 563–571
CrossRef
Google scholar
|
[20] |
Dheeman D S, Henehan G T, Frias J M. Purification and properties of Amycolatopsis mediterranei DSM 43304 lipase and its potential in flavour ester synthesis. Bioresource Technology, 2011, 102(3): 3373–3379
CrossRef
Google scholar
|
[21] |
Wang Y, Zhang D H, Chen N, Zhi G Y. Synthesis of benzyl cinnamate by enzymatic esterification of cinnamic acid. Bioresource Technology, 2015, 198: 256–261
CrossRef
Google scholar
|
[22] |
Kim Y H, Park S. Surveying enantioselectivity of two Candida antarctica-lipase-B homologs towards chiral sec-alcohols. Bulletin of the Korean Chemical Society, 2017, 38(11): 1358–1361
CrossRef
Google scholar
|
[23] |
Liu Z Q, Zheng X B, Zhang S P, Zheng Y G. Cloning, expression and characterization of a lipase gene from the Candida antarctica ZJB09193 and its application in biosynthesis of vitamin A esters. Microbiological Research, 2012, 167(8): 452–460
CrossRef
Google scholar
|
[24] |
Shen J W, Qi J M, Zhang X J, Liu Z Q, Zheng Y G. Significantly increased catalytic activity of Candida antarctica lipase B for the resolution of cis-(±)-dimethyl 1-acetylpiperidine-2,3-dicarboxylate. Catalysis Science & Technology, 2018, 8(18): 4718–4725
CrossRef
Google scholar
|
[25] |
Winkler U K. Glycogen, hyaluronate, and some other polysaccharides greatly enhance the formation of exolipase by Serratia marcescens. Journal of Bacteriology, 1979, 138: 663–670
|
[26] |
Martins A B, da Silva A M, Schein M F, Garcia-Galan C, Záchia Ayub M A, Fernández-Lafuente R, Rodrigues R C. Comparison of the performance of commercial immobilized lipases in the synthesis of different flavor esters. Journal of Molecular Catalysis. B, Enzymatic, 2014, 105: 18–25
CrossRef
Google scholar
|
[27] |
Juturu V, Wu J C. Heterologous protein expression in Pichia pastoris: Latest research progress and applications. ChemBioChem, 2018, 19(1): 7–21
CrossRef
Google scholar
|
[28] |
Eom G T, Lee S H, Song B K, Chung K W, Kim Y W, Song J K. High-level extracellular production and characterization of Candida antarctica lipase B in Pichia pastoris. Journal of Bioscience and Bioengineering, 2013, 116(2): 165–170
CrossRef
Google scholar
|
[29] |
Romdhane I B B, Fendri A, Gargouri Y, Gargouri A, Belghith H. A novel thermoactive and alkaline lipase from Talaromyces thermophilus fungus for use in laundry detergents. Biochemical Engineering Journal, 2010, 53(1): 112–120
CrossRef
Google scholar
|
[30] |
Zheng Y Y, Guo X H, Song N N, Li D C. Thermophilic lipase from Thermomyces lanuginosus: Gene cloning, expression and characterization. Journal of Molecular Catalysis. B, Enzymatic, 2011, 69(3-4): 127–132
CrossRef
Google scholar
|
[31] |
Zhang X F, Yang G Y, Zhang Y, Xie Y, Withers S G, Feng Y. A general and efficient strategy for generating the stable enzymes. Scientific Reports, 2016, 6(1): 33797
CrossRef
Google scholar
|
[32] |
Zhang X F, Ai Y H, Xu Y, Yu X W. High-level expression of Aspergillus niger lipase in Pichia pastoris: Characterization and gastric digestion in vitro. Food Chemistry, 2019, 274: 305–313
CrossRef
Google scholar
|
[33] |
Xie W, Huang M. Immobilization of Candida rugosa lipase onto graphene oxide Fe3O4 nanocomposite: Characterization and application for biodiesel production. Energy Conversion and Management, 2018, 159: 42–53
CrossRef
Google scholar
|
[34] |
Hiol A, Jonzo M D, Rugani N, Druet D, Sarda L, Comeau L C. Purification and characterization of an extracellular lipase from a thermophilic Rhizopus oryzae strain isolated from palm fruit. Enzyme and Microbial Technology, 2000, 26(5-6): 421–430
CrossRef
Google scholar
|
[35] |
Kohno M, Kugimiya W, Hashimoto Y, Morita Y. Purification, characterization, and crystallization of two types of lipase from Rhizopus niveus. Bioscience, Biotechnology, and Biochemistry, 1994, 58(6): 1007–1012
CrossRef
Google scholar
|
[36] |
Florczak T, Daroch M, Wilkinson M C, Bialkowska A, Bates A D, Turkiewicz M, Iwanejko L A. Purification, characterisation and expression in Saccharomyces cerevisiae of LipG7 an enantioselective, cold-adapted lipase from the Antarctic filamentous fungus Geomyces sp. P7 with unusual thermostability characteristics. Enzyme and Microbial Technology, 2013, 53(1): 18–24
CrossRef
Google scholar
|
[37] |
Castro-Ochoa L D, Rodríguez-Gómez C, Valerio-Alfaro G, Oliart Ros R. Screening, purification and characterization of the thermoalkalophilic lipase produced by Bacillus thermoleovorans CCR11. Enzyme and Microbial Technology, 2005, 37(6): 648–654
CrossRef
Google scholar
|
[38] |
Sun Q, Wang H, Zhang H, Luo H, Shi P, Bai Y, Lu F, Yao B, Huang H. Heterologous production of an acidic thermostable lipase with broad-range pH activity from thermophilic fungus Neosartorya fischeri P1. Journal of Bioscience and Bioengineering, 2016, 122(5): 539–544
CrossRef
Google scholar
|
[39] |
Bakir Z B, Metin K. Purification and characterization of an alkali-thermostable lipase from thermophilic Anoxybacillus flavithermus HBB 134. Journal of Microbiology and Biotechnology, 2016, 26(6): 1087–1097
CrossRef
Google scholar
|
[40] |
Sanchez-Carbente M D R, Batista-Garcia R A, Sanchez-Reyes A, Escudero-Garcia A, Morales-Herrera C, Cuervo-Soto L I, French-Pacheco L, Fernández-Silva A, Amero C, Castillo E, Folch-Mallol J L. The first description of a hormone-sensitive lipase from a basidiomycete: Structural insights and biochemical characterization revealed Bjerkandera adusta BaEstB as a novel esterase. MicrobiologyOpen, 2017, 6(4): e00463
CrossRef
Google scholar
|
[41] |
Jallouli R, Parsiegla G, Carriere F, Gargouri Y, Bezzine S. Efficient heterologous expression of Fusarium solani lipase, FSL2, in Pichia pastoris, functional characterization of the recombinant enzyme and molecular modeling. International Journal of Biological Macromolecules, 2017, 94: 61–71
CrossRef
Google scholar
|
[42] |
Zheng X, Chu X, Zhang W, Wu N, Fan Y. A novel cold-adapted lipase from Acinetobacter sp. XMZ-26: Gene cloning and characterisation. Applied Microbiology and Biotechnology, 2011, 90(3): 971–980
CrossRef
Google scholar
|
[43] |
Gricajeva A, Bendikiene V, Kalediene L. Lipase of Bacillus stratosphericus L1: Cloning, expression and characterization. International Journal of Biological Macromolecules, 2016, 92(3): 96–104
CrossRef
Google scholar
|
[44] |
Yang W, He Y, Xu L, Zhang H, Yan Y. A new extracellular thermo-solvent-stable lipase from Burkholderia ubonensis SL-4: Identification, characterization and application for biodiesel production. Journal of Molecular Catalysis. B, Enzymatic, 2016, 126: 76–89
CrossRef
Google scholar
|
[45] |
Bancerz R, Ginalska G. Ginalska G. A novel thermostable lipase from basidiomycete Bjerkandera adusta R59: characterisation and esterification studies. Journal of Industrial Microbiology & Biotechnology, 2007, 34(8): 553–560
CrossRef
Google scholar
|
[46] |
Malekabadi S, Badoei-Dalfard A, Karami Z. Biochemical characterization of a novel cold-active, halophilic and organic solvent-tolerant lipase from B. licheniformis KM12 with potential application for biodiesel production. International Journal of Biological Macromolecules, 2018, 109: 389–398
CrossRef
Google scholar
|
[47] |
Glogauer A, Martini V P, Faoro H, Couto G H, Muller-Santos M, Monteiro R A, Mitchell D A, de Souza E M, Pedrosa F O, Krieger N. Identification and characterization of a new true lipase isolated through metagenomic approach. Microbial Cell Factories, 2011, 10(1): 54
CrossRef
Google scholar
|
[48] |
Ramani K, Chockalingam E, Sekaran G. Production of a novel extracellular acidic lipase from Pseudomonas gessardii using slaughterhouse waste as a substrate. Journal of Industrial Microbiology & Biotechnology, 2010, 37(5): 531–535
CrossRef
Google scholar
|
[49] |
Salihu A, Alam M Z. Solvent tolerant lipases: A review. Process Biochemistry, 2015, 50(1): 86–96
CrossRef
Google scholar
|
[50] |
Dutta Banik S, Nordblad M, Woodley J M, Peters G H. A correlation between the activity of Candida antarctica lipase B and differences in binding free energies of organic solvent and substrate. ACS Catalysis, 2016, 6(10): 6350–6361
CrossRef
Google scholar
|
[51] |
Jin Z, Ntwali J, Han S Y, Zheng S P, Lin Y. Production of flavor esters catalyzed by CALB-displaying Pichia pastoris whole-cells in a batch reactor. Journal of Biotechnology, 2012, 159(1-2): 108–114
CrossRef
Google scholar
|
[52] |
Han S Y, Pan Z Y, Huang D F, Ueda M, Wang X N, Lin Y. Highly efficient synthesis of ethyl hexanoate catalyzed by CALB-displaying Saccharomyces cerevisiae whole-cells in non-aqueous phase. Journal of Molecular Catalysis. B, Enzymatic, 2009, 59(1-3): 168–172
CrossRef
Google scholar
|
[53] |
Shen J W, Qi J M, Zhang X J, Liu Z Q, Zheng Y G. Efficient resolution of cis-(±)-dimethyl 1-acetylpiperidine-2,3-dicarboxylate by covalently immobilized mutant Candida antarctica lipase B in batch and semi-continuous modes. Organic Process Research & Development, 2019, 23(5): 1017–1025
CrossRef
Google scholar
|
[54] |
Zhang X J, Shi P X, Deng H Z, Wang X X, Liu Z Q, Zheng Y G. Biosynthesis of chiral epichlorohydrin using an immobilized halohydrin dehalogenase in aqueous and non-aqueous phase. Bioresource Technology, 2018, 263: 483–490
CrossRef
Google scholar
|
[55] |
Onoja E, Chandren S, Razak F I A, Wahab R A. Enzymatic synthesis of butyl butyrate by Candida rugosa lipase supported on magnetized-nanosilica from oil palm leaves: Process optimization, kinetic and thermodynamic study. Journal of The Taiwan Institute of Chemical Engineers, 2018, 91: 105–118
CrossRef
Google scholar
|
[56] |
Duan X, Liu Y, You X, Jiang Z, Yang S, Yang S. High-level expression and characterization of a novel cutinase from Malbranchea cinnamomea suitable for butyl butyrate production. Biotechnology for Biofuels, 2017, 10(1): 223
CrossRef
Google scholar
|
[57] |
Chen B, Liu H, Guo Z, Huang J, Wang M, Xu X, Zheng L. Lipase-catalyzed esterification of ferulic acid with oleyl alcohol in ionic liquid/isooctane binary systems. Journal of Agricultural and Food Chemistry, 2011, 59(4): 1256–1263
CrossRef
Google scholar
|
[58] |
Chowdary G V, Prapulla S G. Enzymatic synthesis of ethyl hexanoate by transesterification. International Journal of Food Science & Technology, 2003, 38(2): 127–133
CrossRef
Google scholar
|
[59] |
Musa N, Latip W, Abd Rahman R, Salleh A, Mohamad Ali M. Immobilization of an Antarctic Pseudomonas AMS8 lipase for low temperature ethyl hexanoate synthesis. Catalysts, 2018, 8(6): 234
CrossRef
Google scholar
|
/
〈 | 〉 |