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Microbial production of butyl butyrate, a flavor and fragrance compound

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Abstract

Butyl butyrate (BB) has been widely used as a flavor and fragrance compound in the beverage, food, perfume, and cosmetic industries. Currently, BB is produced through two-step processes; butanol and butyrate are first produced and are used as precursors for the esterification reactions to yield BB in the next step. Recently, an alternative process to the current process has been developed by using microorganisms for the one-pot BB production. In the one-pot BB process, alcohol acyl transferases (AATs) and lipases play roles in the esterification of butanol together with their co-substrates butyryl-CoA and butyrate, respectively. In this paper, we review the characteristics of two enzymes including AAT and lipase in the esterification reaction. Also, we review the one-pot processes for BB production by employing the wild-type and engineered Clostridium species and the engineered Escherichia coli strains, with the combination of AATs and lipases.

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References

  • Balbontin C, Gaete-Eastman C, Fuentes L, Figueroa CR, Herrera R, Manriquez D, Latche A, Pech J-C, Moya-León MaA (2010) VpAAT1, a gene encoding an alcohol acyltransferase, is involved in ester biosynthesis during ripening of mountain papaya fruit. J Agr Food Chem 58:5114–5121

  • Beekwilder J, Alvarez-Huerta M, Neef E, Verstappen FW, Bouwmeester HJ, Aharoni A (2004) Functional characterization of enzymes forming volatile esters from strawberry and banana. Plant Physiol 135:1865–1878

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chowdary GV, Prapulla SG (2002) The influence of water activity on the lipase catalyzed synthesis of butyl butyrate by transesterification. Process Biochem 38:393–397

    Article  CAS  Google Scholar 

  • Cumplido-Laso G, Medina-Puche L, Moyano E, Hoffmann T, Sinz Q, Ring L (2012) The fruit ripening-related gene FaAAT2 encodes an acyl transferase involved in strawberry aroma biogenesis. J Exp Bot 63:4275–4290

  • d’Espaux L, Mendez-Perez D, Li R, Keasling JD (2015) Synthetic biology for microbial production of lipid-based biofuels. Curr Opin Chem Biol 29:58–65

    Article  CAS  PubMed  Google Scholar 

  • Defilippi BG, Kader AA, Dandekar AM (2005) Apple aroma: alcohol acyltransferase, a rate limiting step for ester biosynthesis, is regulated by ethylene. Plant Sci 168:1199–1210

    Article  CAS  Google Scholar 

  • Desai RP, Harris LM, Welker NE, Papoutsakis ET (1999) Metabolic flux analysis elucidates the importance of the acid-formation pathways in regulating solvent production by Clostridium acetobutylicum. Metab Eng 1:206–213

    Article  CAS  PubMed  Google Scholar 

  • Dhake KP, Thakare DD, Bhanage BM (2012) Lipase: a potential biocatalyst for the synthesis of valuable flavor and fragrance ester compounds. Flavour Frag J 28:71–83

    Article  CAS  Google Scholar 

  • Emond S, Montanier C, Nicaud J-M, Marty A, Monsan P, André I, Remaud-Siméon M (2010) New efficient recombinant expression system to engineer Candida antarctica lipase B. Appl Environ Microbiol 76:2684–2687

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Fujii T, Yoshimoto H, Nagasawa N, Bogaki T, Tamai Y, Hamachi M (1996) Nucleotide sequences of alcohol acetyltransferase genes from lager brewing yeast, Saccharomyces carlsbergensis. Yeast 12:593–598

    Article  CAS  PubMed  Google Scholar 

  • Fujiwara D, Kobayashi O, Yoshimoto H, Harashima S, Tamai Y (1999) Molecular mechanism of the multiple regulation of the Saccharomyces cerevisiae ATF1 gene encoding alcohol acetyltransferase. Yeast 15:1183–1197

  • Gim GH, Kim SW (2018) Optimization of cell disruption and transesterification of lipids from Botryococcus braunii LB572. Biotechnol Bioprocess Eng 23:550–556

    Article  CAS  Google Scholar 

  • Günther CS, Chervin C, Marsh KB, Newcomb RD, Souleyre EJ (2011) Characterisation of two alcohol acyltransferases from kiwifruit (Actinidia spp.) reveals distinct substrate preferences. Phytochem 72:700–710

    Article  CAS  Google Scholar 

  • Gupta R, Gupta N, Rathi P (2004) Bacterial lipases: an overview of production, purification and biochemical properties. Appl Microbiol Biotechnol 64:763–781

    Article  CAS  PubMed  Google Scholar 

  • Han X, Zhou L (2011) Optimization of process variables in the synthesis of butyl butyrate using acid ionic liquid as catalyst. Chem Eng 172:459–466

    Article  CAS  Google Scholar 

  • Han S-Y, Pan Z-Y, Huang D-F, Ueda M, Wang X-N, Lin Y (2009) Highly efficient synthesis of ethyl hexanoate catalyzed by CALB-displaying Saccharomyces cerevisiae whole-cells in non-aqueous phase. J Mol Catal B Enzym 59:168–172

    Article  CAS  Google Scholar 

  • Haque MA, Hong SY, Hwang CE, Kim SC, Cho KM (2018) Cloning of an organophosphorus hydrolase (opdD) gene of Lactobacillus sakei WCP904 isolated from chlorpyrifos-impregnated kimchi and hydrolysis activities of its gene product for organophosphorus pesticides. Appl Biol Chem 61:643–651

    Article  Google Scholar 

  • Hasan F, Shah AA, Hameed A (2006) Industrial applications of microbial lipases. Enzyme Microb Tech 39:235–251

    Article  CAS  Google Scholar 

  • Horton CE, Bennett GN (2006) Ester production in E. coli and C. acetobutylicum. Enzym Microb Technol 38:937–943

    Article  CAS  Google Scholar 

  • Horton CE, Huang K-X, Bennett GN, Rudolph FB (2003) Heterologous expression of the Saccharomyces cerevisiae alcohol acetyltransferase genes in Clostridium acetobutylicum and Escherichia coli for the production of isoamyl acetate. J Ind Microbiol Biotechnol 30:427–432

    Article  CAS  PubMed  Google Scholar 

  • Inui M, Suda M, Kimura S, Yasuda K, Suzuki H, Toda H, Yamamoto S, Okino S, Suzuki N, Yukawa H (2008) Expression of Clostridium acetobutylicum butanol synthetic genes in Escherichia coli. Appl Microbiol Biotechnol 77:1305–1316

    Article  CAS  PubMed  Google Scholar 

  • Jang Y-S, Lee JY, Lee J, Park JH, Im JA, Eom M-H, Lee J, Lee S-H, Song H, Cho J-H (2012) Enhanced butanol production obtained by reinforcing the direct butanol-forming route in Clostridium acetobutylicum. mBio 3:e00314–12

  • Jenkins RW, Munro M, Nash S, Chuck CJ (2013) Potential renewable oxygenated biofuels for the aviation and road transport sectors. Fuel 103:593–599

    Article  CAS  Google Scholar 

  • Jung S-M, Park Y-C, Park K (2010) Effects of environmental conditions and methanol feeding strategy on lipase-mediated biodiesel production using soybean oil. Biotechnol Bioprocess Eng 15:614–619

    Article  CAS  Google Scholar 

  • Kalscheuer R, Stöveken T, Luftmann H, Malkus U, Reichelt R, Steinbüchel A (2006) Neutral lipid biosynthesis in engineered Escherichia coli: jojoba oil-like wax esters and fatty acid butyl esters. Appl Environ Microbiol 72:1373–1379

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kim J (2017) Surface display of lipolytic enzyme, lipase A and lipase B of Bacillus subtilis on the Bacillus subtilis spore. Biotechnol Bioprocess Eng 22:462–468

  • Kim RJ, Suh MC (2016) The GxSxG motif of Arabidopsis monoacylglycerol lipase (MAGL6 and MAGL8) is essential for their enzyme activities. Appl Biol Chem 59:833–840

    Article  CAS  Google Scholar 

  • Kim D-H, Jeon Y-J, Chung M-J, Seo J-G, Ro Y-T (2017) Complete sequence and gene analysis of a cryptic plasmid pLU4 in Lactobacillus reuteri strain LU4 (KCTC 12397BP). Appl Biol Chem 60:145–153

    Article  CAS  Google Scholar 

  • Kirdi R, Ben Akacha N, Messaoudi Y, Gargouri M (2017) Enhanced synthesis of isoamyl acetate using liquid-gas biphasic system by the transesterification reaction of isoamyl alcohol obtained from fusel oil. Biotechnol Bioprocess Eng 22:413–422

    Article  CAS  Google Scholar 

  • Kruis AJ, Levisson M, Mars AE, van der Ploeg M, Daza FG, Ellena V, Kengen SW, van der Oost J, Weusthuis RA (2017) Ethyl acetate production by the elusive alcohol acetyltransferase from yeast. Metab Eng 41:92–101

    Article  CAS  PubMed  Google Scholar 

  • Kumar V, Kumar R, Rawat D, Nanda M (2018) Synergistic dynamics of light, photoperiod and chemical stimulants influences biomass and lipid productivity in Chlorella singularis (UUIND5) for biodiesel production. Appl Biol Chem 61:7–13

    Article  CAS  Google Scholar 

  • Langrand G, Rondot N, Triantaphylides C, Baratti J (1990) Short chain flavour esters synthesis by microbial lipases. Biotechnol Lett 12:581–586

    Article  CAS  Google Scholar 

  • Layton DS, Trinh CT (2014) Engineering modular ester fermentative pathways in Escherichia coli. Metab Eng 26:77–88

    Article  CAS  PubMed  Google Scholar 

  • Layton DS, Trinh CT (2016a) Expanding the modular ester fermentative pathways for combinatorial biosynthesis of esters from volatile organic acids. Biotechnol Bioeng 113:1764–1776

    Article  CAS  PubMed  Google Scholar 

  • Layton DS, Trinh CT (2016b) Microbial synthesis of a branched-chain ester platform from organic waste carboxylates. Metab Eng Commun 3:245–251

    Article  PubMed  PubMed Central  Google Scholar 

  • Lee J, Jang Y-S, Choi SJ, Im JA, Song H, Cho JH, Papoutsakis ET, Bennett GN, Lee SY (2012) Metabolic engineering of Clostridium acetobutylicum ATCC 824 for isopropanol-butanol-ethanol fermentation. Appl Environ Microbiol 78:1416–1423

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li F, Wu X, Lam P, Bird D, Zheng H, Samuels L, Jetter R, Kunst L (2008) Identification of the wax ester synthase/acyl-coenzyme A: diacylglycerol acyltransferase WSD1 required for stem wax ester biosynthesis in Arabidopsis. Plant Physiol 148:97–107

  • Li W, Cui D-Y, Wang J-H, Liu X-E, Xu J, Zhou Z, Zhang C-Y, Chen Y-F, Xiao D-G (2018) Overexpression of different alcohol acetyltransferase genes with BAT2 deletion in Saccharomyces cerevisiae affects acetate esters and higher alcohols. Eur Food Res Technol 244:555–564

  • Liu T, Zhang J (2018) High-level expression and characterization of Aspergillus niger ATCC 1015 xylanase B in Komagataella phaffii. Appl Biol Chem 61:373–381

  • Lozano P, Pérez-Marın A, De Diego T, Gomez D, Paolucci-Jeanjean D, Belleville M, Rios G, Iborra J (2002) Active membranes coated with immobilized Candida antarctica lipase B: preparation and application for continuous butyl butyrate synthesis in organic media. J Membr Biol 201:55–64

    Article  CAS  Google Scholar 

  • Mancheno JM, Pernas MA, Martınez MJ, Ochoa B, Rúa ML, Hermoso JA (2003) Structural insights into the lipase/esterase behavior in the Candida rugosa lipases family: crystal structure of the lipase 2 isoenzyme at 1.97 Å resolution. J Mol Biol 332:1059–1069

    Article  CAS  PubMed  Google Scholar 

  • Martins AB, Friedrich JL, Cavalheiro JC, Garcia-Galan C, Barbosa O, Ayub MA, Fernandez-Lafuente R, Rodrigues RC (2013) Improved production of butyl butyrate with lipase from Thermomyces lanuginosus immobilized on styrene–divinylbenzene beads. Bioresour Technol 134:417–422

    Article  CAS  PubMed  Google Scholar 

  • Matte CR, Bordinhão C, Poppe JK, Rodrigues RC, Hertz PF, Ayub MA (2016) Synthesis of butyl butyrate in batch and continuous enzymatic reactors using Thermomyces lanuginosus lipase immobilized in Immobead 150. J Mol Catal B Enzym 127:67–75

    Article  CAS  Google Scholar 

  • Menendez-Bravo S, Comba S, Gramajo H, Arabolaza A (2017) Metabolic engineering of microorganisms for the production of structurally diverse esters. Appl Microbiol Biotechnol 101:3043–3053

    Article  CAS  PubMed  Google Scholar 

  • Nancolas B, Bull ID, Stenner R, Dufour V, Curnow P (2017) Saccharomyces cerevisiae Atf1p is an alcohol acetyltransferase and a thioesterase in vitro. Yeast 34:239–251

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Noh HJ, Woo JE, Lee SY, Jang Y-S (2018) Metabolic engineering of Clostridium acetobutylicum for the production of butyl butyrate. Appl Microbiol Biotechnol 102:8319–8327

    Article  CAS  PubMed  Google Scholar 

  • Olias JM, Sanz C, Rios J, Perez AG (1995) Substrate specificity of alcohol acyltransferase from strawberry and banana fruits. ACS Symp Ser (USA) 596:134–141

    Article  CAS  Google Scholar 

  • Park S-C, Chang W-J, Lee S-M, Kim Y-J, Koo Y-M (2005) Lipase-catalyzed transesterification in several reaction systems: an application of room temperature lonic liquids for bi-phasic production of n-butyl acetate. Biotechnol Bioprocess Eng 10:99–102

    Article  CAS  Google Scholar 

  • Park S-J, Kim D-H, Yoo J, Hwang EY, Shin M-S, Lee N-T, Cho I-R, Kang H-G, Kim Y-J, Park S, Kim Y-W (2017a) Detection of organophosphate bound butyrylcholinesterase using a monoclonal antibody. Appl Biol Chem 60:233–240

    Article  CAS  Google Scholar 

  • Park S, Kim K, Han S-I, Kim EJ, Choi Y-E (2017b) Organic solvent-free lipid extraction from wet Aurantiochytrium sp. biomass for co-production of biodiesel and value-added products. Appl Biol Chem 60:101–108

    Article  CAS  Google Scholar 

  • Rodriguez GM, Tashiro Y, Atsumi S (2014) Expanding ester biosynthesis in Escherichia coli. Nat Chem Biol 10:259–265

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Saerens SM, Delvaux FR, Verstrepen KJ, Thevelein JM (2010) Production and biological function of volatile esters in Saccharomyces cerevisiae. Microb Biotechnol 3:165–177

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Salas JJ (2004) Characterization of alcohol acyltransferase from olive fruit. J Agr Food Chem 52:3155–3158

    Article  CAS  Google Scholar 

  • Santos JC, Nunes GF, Moreira AB, Perez VH, de Castro HF (2007) Characterization of Candida rugosa lipase immobilized on poly (N-methylolacrylamide) and its application in butyl butyrate synthesis. Chem Eng Technol 30:1255–1261

    Article  CAS  Google Scholar 

  • Schneiderbanger H, Koob J, Poltinger S, Jacob F, Hutzler M (2016) Gene expression in wheat beer yeast strains and the synthesis of acetate esters. J I Brewing 122:403–411

    Article  CAS  Google Scholar 

  • Selvam K, Vishnupriya B, Maanvizhi M (2013) Enzymatic synthesis of fragrance ester by lipase from marine Actinomycetes for textile industry. Int J Eng Adv Technol 3:91–96

  • Seo S-O, Wang Y, Lu T, Jin Y-S, Blaschek HP (2017) Characterization of a Clostridium beijerinckii spo0A mutant and its application for butyl butyrate production. Biotechnol Bioeng 114:106–112

  • Sudheer PDVN, Yun J, Chauhan S, Kang TJ, Choi K-Y (2017) Screening, expression, and characterization of Baeyer-Villiger monooxygenases for the production of 9-(nonanoyloxy)nonanoic acid from oleic acid. Biotechnol Bioprocess Eng 22:717–724

    Article  CAS  Google Scholar 

  • Tamalampudi S, Talukder MMR, Hama S, Tanino T, Suzuki Y, Kondo A, Fukuda H (2007) Development of recombinant Aspergillus oryzae whole-cell biocatalyst expressing lipase-encoding gene from Candida antarctica. Appl Microbiol Biotechnol 75:387–395

    Article  CAS  PubMed  Google Scholar 

  • van den Berg C, Heeres AS, van der Wielen LAM, Straathof AJJ (2013) Simultaneous clostridial fermentation, lipase-catalyzed esterification, and ester extraction to enrich diesel with butyl butyrate. Biotechnol Bioeng 110:137–142

    Article  CAS  PubMed  Google Scholar 

  • Van Laere SD, Saerens SM, Verstrepen KJ, Van Dijck P, Thevelein JM, Delvaux FR (2008) Flavour formation in fungi: characterisation of KlAtf, the Kluyveromyces lactis orthologue of the Saccharomyces cerevisiae alcohol acetyltransferases Atf1 and Atf2. Appl Microbiol Biotechnol 78:783–792

    Article  CAS  PubMed  Google Scholar 

  • Varma MN, Madras G (2008) Kinetics of synthesis of butyl butyrate by esterification and transesterification in supercritical carbon dioxide. J Chem Technol Biotechnol 83:1135–1144

    Article  CAS  Google Scholar 

  • Wiesenborn DP, Rudolph FB, Papoutsakis ET (1988) Thiolase from Clostridium acetobutylicum ATCC 824 and its role in the synthesis of acids and solvents. Appl Environ Microbiol 54:2717–2722

    CAS  PubMed  PubMed Central  Google Scholar 

  • Woo JE, Lee SY, Jang Y-S (2018) Effects of nutritional enrichment on acid production from degenerated (non-solventogenic) Clostridium acetobutylicum strain M5. Appl Biol Chem 61:469–472

    Article  CAS  Google Scholar 

  • Xin F, Basu A, Yang K-L, He J (2016) Strategies for production of butanol and butyl-butyrate through lipase-catalyzed esterification. Bioresour Technol 202:214–219

    Article  CAS  PubMed  Google Scholar 

  • Yeom SH, Go YW (2018) Optimization of a novel two-step process comprising re-esterification and transesterification in a single reactor for biodiesel production using waste cooking oil. Biotechnol Bioprocess Eng 23:432–441

    Article  CAS  Google Scholar 

  • Yeon YJ, Park H-Y, Park K, Park HJ, Yoo YJ (2016) Structural basis for the substrate specificity of 3-hydroxybutyrate dehydrogenase. Biotechnol Bioprocess Eng 21:364–372

    Article  CAS  Google Scholar 

  • Yu Y, Wu D, Liu C, Zhao Z, Yang Y, Li Q (2012) Lipase/esterase-catalyzed synthesis of aliphatic polyesters via polycondensation: a review. Process Biochem 47:1027–1036

    Article  CAS  Google Scholar 

  • Zabetakis I, Holden MA (1997) Strawberry flavour: analysis and biosynthesis. J Sci Food Agric 74:421–434

    Article  CAS  Google Scholar 

  • Zhang C-Y, Liu Y-L, Qi Y-N, Zhang J-W, Dai L-H, Lin X, Xiao D-G (2013) Increased esters and decreased higher alcohols production by engineered brewer’s yeast strains. Eur Food Res Technol 236:1009–1014

    Article  CAS  Google Scholar 

  • Zhang J, Zhang C, Qi Y, Dai L, Ma H, Guo X, Xiao D (2014) Acetate ester production by Chinese yellow rice wine yeast overexpressing the alcohol acetyltransferase-encoding gene ATF2. Genet Mol Res 13:9735–9746

  • Zhang ZT, Taylor S, Wang Y (2017) In situ esterification and extractive fermentation for butyl butyrate production with Clostridium tyrobutyricum. Biotechnol Bioeng 114:1428–1437

    Article  CAS  PubMed  Google Scholar 

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Funding

This work was supported by a grant from the Ministry of Science and ICT (MSIT) through the National Research Foundation (NRF) of Korea (NRF-2016R1D1A3B04933184). SYL was supported by the Technology Development Program to Solve Climate Changes on Systems Metabolic Engineering for Biorefineries from the MSIT through the NRF of Korea (NRF-2012M1A2A2026556 and NRF-2012M1A2A2026557).

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  1. Department of Agricultural Chemistry and Food Science Technology, Division of Applied Life Science (BK21 Plus Program), Institute of Agriculture & Life Science (IALS), Gyeongsang National University, Jinju, Republic of Korea

    Hyeon Ji Noh & Yu-Sin Jang

  2. Department of Chemical and Biomolecular Engineering (BK21 Plus Program), BioProcess Engineering Research Center, Institute for the BioCentury, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Republic of Korea

    Sang Yup Lee

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  1. Hyeon Ji Noh
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Noh, H.J., Lee, S.Y. & Jang, YS. Microbial production of butyl butyrate, a flavor and fragrance compound. Appl Microbiol Biotechnol 103, 2079–2086 (2019). https://doi.org/10.1007/s00253-018-09603-z

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