Abstract
Many flavoring agents on the market are extracted from natural sources or synthesized chemically. Due to the disadvantages of both methods, biotechnology is becoming a promising alternative. In this study, short chain ethyl esters with fruity notes were biosynthesized in UHT whole milk via coupling ethanolic fermentation with lipase (Palatase®) transesterification. Kluyveromyces marxianus, Lactobacillus fermentum and Lb. paracasei were used for fermentation. Milk fat was esterified with in situ produced ethanol by adding lipase at 0, 8 and 24 h of fermentation. Viable cell counts and pH were monitored during 48 h fermentation period. Flavor active ethyl esters, ethanol and free fatty acids were analyzed using headspace SPME-GC. Free fatty acid levels were lower in K. marxianus samples than lactobacilli. K. marxianus produced higher amounts of ethanol and esters than lactic acid bacteria. Viable cell counts decreased after lipase application at 0 and 8 h, possibly due to fatty acid production. Addition of lipase at 24 h resulted in improved cell counts as well as ethanol and ester production in the case of K. marxianus. This study demonstrated that fermenting milk with alcohol producing cultures in conjunction with lipase application can be an alternative to artificial flavorings in fermented milks.
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References
Alzeer J, Abou Hadeed K (2016) Ethanol and its Halal status in food industries. Trends Food Sci Technol 58:14–20. https://doi.org/10.1016/j.tifs.2016.10.018
Annan NT, Poll L, Sefa-Dedeh S, Plahar WA, Jakobsen M (2003) Volatile compounds produced by Lactobacillus fermentum, Saccharomyces cerevisiae and Candida krusei in single starter culture fermentations of Ghanaian maize dough. J Appl Microbiol 94(3):462–474. https://doi.org/10.1046/j.1365-2672.2003.01852.x
Basso LC, Basso TO, Rocha SN (2011) Ethanol production in Brazil: the industrial process and its impact on yeast fermentation. In: Bernardes MAS (ed) Biofuel production-recent developments and prospects. IntechOpen, London, pp 85–100. https://doi.org/10.5772/17047
Condurso C, Verzera A, Romeo V, Ziino M, Conte F (2008) Solid-phase microextraction and gas chromatography mass spectrometry analysis of dairy product volatiles for the determination of shelf-life. Int Dairy J 18(8):819–825. https://doi.org/10.1016/j.idairyj.2007.12.005
Danielsen M, Wind A (2003) Susceptibility of Lactobacillus spp. to antimicrobial agents. Int J Food Microbiol 82(1):1–11. https://doi.org/10.1016/S0168-1605(02)00254-4
Delgado P, Sanz MT, Beltrán S, Núñez LA (2010) Ethyl lactate production via esterification of lactic acid with ethanol combined with pervaporation. Chem Eng J 165(2):693–700. https://doi.org/10.1016/j.cej.2010.10.009
Hamme V, Sannier F, Piot JM, Didelot S, Bordenave-Juchereau S (2009) Crude goat whey fermentation by Kluyveromyces marxianus and Lactobacillus rhamnosus: contribution to proteolysis and ACE inhibitory activity. J Dairy Res 76(2):152–157. https://doi.org/10.1017/S0022029908003877
Jensen RG, Galluzzo DR, Bush VJ (1990) Selectivity is an important characteristic of lipases (acylglycerol hydrolases). Biocatalysis 3(4):307–316. https://doi.org/10.3109/10242429008992074
Khurana HK, Kanawjia SK (2007) Recent trends in development of fermented milks. Curr Nutr Food Sci 3(1):91–108. https://doi.org/10.2174/1573401310703010091
Kurtovic I, Marshall SN, Miller MR, Zhao X (2011) Flavour development in dairy cream using fish digestive lipases from Chinook salmon (Oncorhynchus tshawytscha) and New Zealand hoki (Macruronus novaezealandiae). Food Chem 127(4):1562–1568. https://doi.org/10.1016/j.foodchem.2011.02.018
Leclercq-Perlat MN, Corrieu G, Spinnler HE (2007) Controlled production of Camembert-type cheeses: part III role of the ripening microflora on free fatty acid concentrations. J Dairy Res 74(2):218–225. https://doi.org/10.1017/S0022029906002329
Liu SQ, Holland R, Crow VL (2003) Synthesis of ethyl butanoate by a commercial lipase in aqueous media under conditions relevant to cheese ripening. J Dairy Res 70:359–363. https://doi.org/10.1017/S0022029903006290
Liu SQ, Holland R, Crow VL (2004) Esters and their biosynthesis in fermented dairy products: a review. Int Dairy J 14(11):923–945. https://doi.org/10.1016/j.idairyj.2004.02.010
Liu SQ, Crow VL, Holland R (2009) Production of natural fruity flavour in dairy foods. Nutr Food Sci 39:483–489. https://doi.org/10.1108/00346650910992132
Liu SQ, Lee HY, Yu B, Curran P, Sun J (2013) Bioproduction of natural isoamyl esters from coconut cream as catalysed by lipases. J Food Res 2(2):157–166. https://doi.org/10.5539/jfr.v2n2p157
Longo MA, Sanromán MA (2006) Production of food aroma compounds: microbial and enzymatic methodologies. Food Technol Biotechnol 44(3):335–353
Löser C, Urit T, Stukert A, Bley T (2013) Formation of ethyl acetate from whey by Kluyveromyces marxianus on a pilot scale. J Biotechnol 163(1):17–23. https://doi.org/10.1016/j.jbiotec.2012.10.009
Lu Y, Putra SD, Liu SQ (2018) A novel non-dairy beverage from durian pulp fermented with selected probiotics and yeast. Int J Food Microbiol 265:1–8. https://doi.org/10.1016/j.ijfoodmicro.2017.10.030
Mital BK, Garg SK (1992) Acidophilus milk products: manufacture and therapeutics. Food Rev Int 8(3):347–389. https://doi.org/10.1080/87559129209540946
Naaber P, Smidt I, Štšepetova J, Brilene T, Annuk H, Mikelsaar M (2004) Inhibition of Clostridium difficile strains by intestinal Lactobacillus species. J Med Microbiol 53(6):551–554. https://doi.org/10.1099/jmm.0.45595-0
Østlie HM, Helland MH, Narvhus JA (2003) Growth and metabolism of selected strains of probiotic bacteria in milk. Int J Food Microbiol 87(1–2):17–27. https://doi.org/10.1016/s0168-1605(03)00044-8
Quigley L, O'Sullivan O, Stanton C, Beresford TP, Ross RP, Fitzgerald GF, Cotter PD (2013) The complex microbiota of raw milk. FEMS Microbiol Rev 37(5):664–698. https://doi.org/10.1111/1574-6976.12030
Rampelotto PH (2010) Resistance of microorganisms to extreme environmental conditions and its contribution to astrobiology. Sustainability 2(6):1602–1623. https://doi.org/10.3390/su2061602
Ricke SC (2003) Perspectives on the use of organic acids and short chain fatty acids as antimicrobials. J Poult Sci 82(4):632–639. https://doi.org/10.1093/ps/82.4.632
Sun J, Liu SQ (2015) Ester synthesis in aqueous media by lipase: alcoholysis, esterification and substrate hydrophobicity. J Food Biochem 39(1):11–18. https://doi.org/10.1111/jfbc.12104
Sun J, Jiang Y, Zhou L, Gao J (2010) Optimization and kinetic study of immobilized lipase-catalyzed synthesis of ethyl lactate. Biocatal Biotransform 28(4):279–287. https://doi.org/10.3109/10242422.2010.501893
Sun J, Yu B, Curran P, Liu SQ (2012) Optimisation of flavour ester biosynthesis in an aqueous system of coconut cream and fusel oil catalysed by lipase. Food Chem 135(4):2714–2720. https://doi.org/10.1016/j.foodchem.2012.06.119
Sun J, Lim Y, Liu SQ (2013) Biosynthesis of flavor esters in coconut cream through coupling fermentation and lipase-catalyzed biocatalysis. Eur J Lipid Sci Technol 115(10):1107–1114. https://doi.org/10.1002/ejlt.201300144
Toh M, Liu SQ (2017) Impact of coculturing Bifidobacterium animalis subsp. lactis HN 019 with yeasts on microbial viability and metabolite formation. J Appl Microbiol 123(4):956–968. https://doi.org/10.1111/jam.13571
Ugur Nigiz F, Durmaz Hilmioglu N (2016) Green solvent synthesis from biomass based source by biocatalytic membrane reactor. Int J Energy Res 40(1):71–80. https://doi.org/10.1002/er.3319
Wu WH, Hung WC, Lo KY, Chen YH, Wan HP, Cheng KC (2016) Bioethanol production from taro waste using thermo-tolerant yeast Kluyveromyces marxianus K21. Bioresour Technol 201:27–32. https://doi.org/10.1016/j.biortech.2015.11.015
Zhang XM, Ai NS, Wang J, Tong LJ, Zheng FP, Sun BG (2016) Lipase-catalyzed modification of the flavor profiles in recombined skim milk products by enriching the volatile components. J Dairy Sci 99(11):8665–8679. https://doi.org/10.3168/jds.2015-10773
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It is a great pleasure for us to thank National University of Singapore (NUS) particularly members of the Chemistry Department for providing laboratory and supporting this research. The support from Shiraz University is also sincerely appreciated.
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MTG and MHE suggested the subject of study and supervised it; SQL designed and led the study; MSZ performed the experiments, analyzed the data and wrote the manuscript; MT provided the materials and directed the experiments implementation and data analysis; all authors revised and commented on the manuscript.
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Shojaei Zinjanab, M., Golmakani, M.T., Eskandari, M.H. et al. Natural flavor biosynthesis by lipase in fermented milk using in situ produced ethanol. J Food Sci Technol 58, 1858–1868 (2021). https://doi.org/10.1007/s13197-020-04697-8
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DOI: https://doi.org/10.1007/s13197-020-04697-8