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Trends in Food Science & Technology 69 (2017) 95e105

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Trends in Food Science & Technology


journal homepage: http://www.journals.elsevier.com/trends-in-food-science-
and-technology

Review

A review on enzymatic synthesis of aromatic esters used as flavor


ingredients for food, cosmetics and pharmaceuticals industries
Amanda Gomes Almeida SA, Alessandra Cristina de Meneses,
bora de Oliveira*
Pedro Henrique Hermes de Araújo, De
polis, Santa Catarina, Brazil
Department of Chemical Engineering and Food Engineering, Federal University of Santa Catarina, Floriano

a r t i c l e i n f o a b s t r a c t

Article history: Background: Many sectors of industry, mainly food, cosmetics and pharmaceutics, have increased their
Received 14 July 2017 interest in esters due to their flavor property. Flavor esters that possess an aromatic ring in their mo-
Received in revised form lecular structure are also known as aromatic esters. These esters are widely found in nature (fruits and
12 September 2017
plants) and the synthetic (i.e. via chemical) and natural routes (i.e. via direct extraction from nature or via
Accepted 13 September 2017
biotechnology) are suitable for their biocatalysis.
Available online 20 September 2017
Scope and approach: In this context, from the industrial point of view, enzyme-catalyzed reactions are
the most economical approach to reach final green products with no toxicity and no harm to human
Keywords:
Flavor
health. The present article gives an overview of the aromatic esters synthesis, considering the main
Biocatalysis effects in the reaction media conditions and enzymes used. This review also describes applied trends in
Enzyme enzymatic-catalyzed reactions, pointing alternatives to production, like ultrasound-assisted reactions
Lipase and process optimization of aromatic esters. Furthermore, this work presents perspectives concerning
Esters the biological potential of these esters and recent advances in their encapsulation.
Encapsulation Key findings and conclusions: Lipases play an important role in the aromatic esters production, with
several advantages over synthetic route. Lipase-catalyzed reactions usually follows Ping-Pong Bi-Bi or
ternary complex (order Bi-Bi) mechanism. The study of the process parameters and their interaction are
very important to understand the system optimization and achieve the maximum reaction yield to scale
up. Aromatic esters can present some biological activities, in addition to their fragrances, which increases
the interest in the encapsulation of these compounds.
© 2017 Elsevier Ltd. All rights reserved.

1. Introduction Aromatic esters are flavor esters that have an aromatic ring in
their molecular structure. These esters are usually obtained via
Flavor esters are significant and versatile compounds, also direct extraction from plant or fruit source; however, disadvantages
known as important ingredients in food, beverages, cosmetics, make this technique inadequate for industrial applications
pharmaceuticals, chemicals and personal care products, like per- including the seasonal and climatic dependency of the source, be-
fumes, body lotions, face creams, shampoos, soaps, shower-shaving sides the low yield and high production costs needed for extraction
gels and other toiletries, due to their flavor and fragrance properties and purification (Badgujar, Sasaki, & Bhanage, 2015; Gao, Wu, Chen,
(Akacha & Gargouri, 2015; Badgujar, Pai, & Bhanage, 2016; Li, Sun, & Fan, 2016; Gumel & Annuar, 2016; Santos, Zabot, Meireles, &
Li, & Liu, 2014; Shintre, Ghadge, & Sawant, 2002; N. C. A.; Silva, Mazutti, 2016).
Miranda, Bolina, & Silva, 2014; Todero, Bassi, Lage, & Corradini, The chemical production of aromatic esters also possesses
2015). several drawbacks and environmental impacts in the production
and purification processes, such as the use of hazardous chemicals
and catalysts, toxic solvents and high temperature and pressure.
Moreover, chemical synthesis presents high costs due to the lack of
* Corresponding author. Universidade Federal de Santa Catarina (UFSC),
rio
Departamento Engenharia Química e Engenharia de Alimentos (EQA), Laborato substrate selectivity and by-products removal, long reaction times,
de Controle de Processos, Campus Trindade, Cx. Postal 476, 88010-970 excessive consumption of energy and possible corrosion of the
polis, SC, Brazil.
Floriano equipment. Furthermore, the final products cannot be legally
E-mail address: debora.oliveira@ufsc.br (D. Oliveira).

https://doi.org/10.1016/j.tifs.2017.09.004
0924-2244/© 2017 Elsevier Ltd. All rights reserved.
96  et al. / Trends in Food Science & Technology 69 (2017) 95e105
A.G.A. SA

labeled as natural and the synthesized esters contain traces of toxic flavoring agents. European Commission, Food and Drug Adminis-
impurities, which may result in humans’ health complications. tration (FDA) and International Joint FAO/WHO Expert Committee
Therefore, all these factors make the process industrially disad- on Food Additives (JECFA) approve these esters for food additives
vantageous (Khan & Rathod, 2015; Manan, Rahman, Marzuki, & (COF, 2000; FDA, 2013; JECFA, 2001).
Mahat, 2016; Mohamad, Buang, Mahat, & Lok, 2015; M. J. A.; The global market for flavors and fragrances was valued at $ 26.0
Silva, Loss, Laroque, & Lerin, 2015; Wang, Zhang, Chen, & Zhi, 2015). billion in 2015. This market should increase from $ 27.1 billion in
Biotechnology is a technique that uses living organisms (or parts 2016 to $ 37.0 billion in 2021 at a compound annual growth rate
of them) to make or modify products, to improve plants and ani- (CAGR) of 6.4% and grow to $ 33.5 billion by 2019 (BBC, 2016).
mals or to develop microorganisms for specific uses (Bicas, Silva,
Dionísio, & Pastore, 2010). Biotechnological transformations (i.e.
3. Enzymes
microbial and enzymatic routes) are employed in ester production
and among various biotechnological processes, the lipase-catalyzed
Enzymes are non-toxic biocatalysts that accelerate the rate of
reaction is economically viable for flavor and aromatic esters syn-
reactions and are highly versatile in the catalysis of various types of
thesis (Garlapati, Kumari, Mahapatra, & Banerjee, 2013; Stencel &
reactions using mild conditions. Enzymes have attracted significant
Leadbeater, 2014). Biocatalysis applied in ester production is a
attention due to their high specificity, high chemo-, regio-, and
useful and promising alternative green tool, which offers several
stereo-selectivity, ease of processing, broad substrate array and
advantages, such as high specificity and chemo-, regio- and stereo-
ability to succeed organic transformations in various reaction me-
selectivity, high yields in mild reaction conditions (low tempera-
dia (Badgujar & Bhanage, 2015; Gryglewicz, Jadownicka, &
ture and pressure), reduction of by-products formation, biocatalyst
Czerniak, 2000; S. Sharma & Kanwar, 2014). Among the enzymes,
reusability, low energy consumption and reduction of the overall
lipases are important because of the large number of reactions
production costs (Ferraz, Possebom, Alvez, & Cansian, 2015; Kuo,
which they can catalyze in organic systems, high stability, versa-
Chen, Chen, & Liu, 2014). In addition, aromatic esters produced
tility and low commercial cost (Giunta, Sechi, & Solinas, 2015;
by microbial or enzymatic methods are labeled as natural in
Jakovetic, Lukovic, Boskovi
c-Vragolovic, & Bezbradica, 2013;
accordance with the United States and European Legislations,
Paroul, Grzegozeski, Chiaradia, & Treichel, 2011).
thereby satisfying the consumer trend towards natural products in
The global market for industrial enzymes reached nearly $ 4.9
various industries (Leszczak & Tran-Minh, 1998; Tomke & Rathod,
billion in 2015. This market should increase from $ 5.0 billion in
2015; Vanin, Orlando, Piazza, & Puton, 2014).
2016 to $ 6.3 billion in 2021 at a compound annual growth rate
This article presents an overview of the current state of the art of
(CAGR) of 4.7% (BBC, 2017).
aromatic esters production by enzymatic route, providing knowl-
edge about the latest advances and most relevant process param-
eters related to their synthesis. This work also reviews the recent 3.1. Lipases
techniques used, optimization and kinetics aiming to improve the
reaction yield and scale up. Furthermore, this review presents Lipases (triacylglycerol hydrolases, EC 3.1.1.3) play an important
perspectives concerning biological potential in ester production, role in organic synthesis and flavor biotechnology (Dubal, Tilkari,
modern progress in aromatic esters encapsulation and the future Momin, & Borkar, 2008; Wu, Qi, Wang, & Su, 2014). These en-
trends, challenges and prospects in this field. zymes are responsible for the hydrolysis of lipids to fatty acids and
glycerol, and possess the ability to catalyze several reactions, such
2. Bioflavors as esterification (alcohol and carboxylic acid), transesterification
(ester and alcohol), interesterification (ester and acid) and transfer
Flavors are composed of different organic chemicals, such as of acyl groups from esters to other nucleophiles (e.g. amines and
hydrocarbons, alcohols, aldehydes, ketones, acids, esters or lac- thiols) (Hoang & Matsuda, 2016; Horchani, Salem, Zarai, & Sayari,
tones. The low volatility and low molecular weight, usually lower 2010; Mendes, Castro, & Giordano, 2014; Narwal, Saun, Dogra, &
than 400 Da, are responsible for a range of sensorial sensations Gupta, 2016; Paludo, Alves, Altmann, & Ayub, 2015; Tomke &
attributed to the flavors (Longo & Sanroma n, 2006). Rathod, 2015).
Flavor and aromatic esters are widely found in nature and confer Lipases are abundant in nature and found in multiple organisms,
pleasant organoleptic impact attributes, including fruity, floral, but yeast and fungi are the main sources of lipases for industrial
spicy, creamy or nutty aromas (Berger, 2009; Gao et al., 2016; Yadav applications. Most commercially important lipase-producing yeasts
& Dhoot, 2009). These properties make possible a great variety of belong to the class of ascomycetes like Candida sp. Novozymes®
applications in the food sector in many beverages, candies, jellies, (Denmark), DuPont® (United States) and Roche® (Switzerland) are
jams, wines and dairy products (Akacha & Gargouri, 2015; Białecka- the main companies for the production and commercialization of
Florjan czyk, Krzyczkowska, Stolarzewicz, & Kapturowska, 2012) lipases (Gupta, Kumari, Syal, & Singh, 2015).
and also in the cosmetic industry, as fragrances in perfumes, de- The practical use of homogeneous biocatalysis (free enzyme)
odorants, creams and soaps and flavors in lip cosmetics. However, has some disadvantages on industrial process economics because
besides the fragrance they can also have others properties, such as of low solvent, thermal, mechanical and operational stabilities and
emollient, surfactant and antioxidant, which make possible their no recyclability, which leads to high production costs. To overcome
application in many formulations of creams, shampoos and anti- these limitations, efforts have been taken by researchers to
aging creams (Becker, Bergfeld, Belsito, & Hill, 2012; Khan & develop several advance skillful immobilization techniques
Rathod, 2015). (Badgujar & Bhanage, 2015; Ferraz et al., 2015; S. Sharma &
The rapid absorption, metabolic excretion, low level of use and Kanwar, 2014), which offer many process advantages: lower pro-
the lack of significant genotoxic and methanogenic potential make duction cost, increased activity, specificity and selectivity,
the aromatic esters Generally Recognized as Safe (GRAS) for use as improved structural stability, reduction of inhibition, ease of sep-
flavor ingredient since 1965 by the Flavor and Extract Manufac- aration, recovery and further reuse of the biocatalyst due to the
turers Association (FEMA) Expert Panel (Adams, Cohen, Doull, & heterogeneous characteristic (Bansode & Rathod, 2014; Dhake,
Feron, 2005a). Furthermore, some international regulations deter- Tambade, Qureshi, & Singhal, 2011; Kuo et al., 2014; Narwal
mine that aromatic esters have no safety concerns when used as et al., 2016).
 et al. / Trends in Food Science & Technology 69 (2017) 95e105
A.G.A. SA 97

4. Aromatic esters Bhatia, & Bruze, 2015, 2016; Adams, Cohen, Doull, & Feron, 2004;
Adams, Cohen, Doull, & Feron, 2005b; Adams et al., 2005a;
Flavor esters that possess an aromatic ring in the molecular Belsito, Bickers, Bruze, & Calow, 2012; McGginty, Letizia, & Api,
structure are also known as aromatic esters. These esters are 2012; McGinty, Letizia, & Api, 2012).
broadly present in nature and confer pleasant organoleptic attri- The biotechnological production of these aromatic esters is an
butes, such as fruity and floral smell and taste, which make possible alternative to natural source and has the possibility of scale up to
a great variety of applications in the food, pharmaceuticals and industrial applications. Some studies have been made in this area,
cosmetic industries (Berger, 2009; Dhake, Deshmukh, Wagh, & with different immobilized lipases as the heterogeneous bio-
Singhal, 2012; Gao et al., 2016; Yadav & Dhoot, 2009). The main catalysts in the synthesis of benzyl acetate (Garlapati et al., 2013),
aromatic esters are benzyl, cresyl, anisyl, eugenyl, cinnamyl, phe- cinnamyl acetate (Badgujar et al., 2015) and 2-methyl benzyl ace-
nethyl, benzoate and cinnamate esters. The applications and nat- tate (Dhake et al., 2012). Tables 2e6 present the main reaction
ural occurrences of these esters are shown in Table 1 (Api, Belsito, parameters for the synthesis of benzyl, cresyl, anisyl, eugenyl,

Table 1
Applications and natural occurrences of aromatic esters.

Aromatic Ester Application in Food, Flavour-Fragance Natural occurrence in plants and fruits

Benzyl acetate Jasmine, green apple, strawberry, banana, orchid Acacia farnesiana, Hyacinthus sp., Jasminum

Benzyl propionate Almond, apple, banana, coconut, grape, cherry, Michelia champaca, Prunus species, Tanacetum
strawberry, plum parthenium and melon

Benzyl butyrate Cherry, apple, berry, grape, plum, pear, peach, papaya, Spondias mombins, Osmanthus fragrans, Vasconcellea
pineapple, raspberry, orchid pubescens, Grape cherimoya, Michelia champaca

Benzyl benzoate Apple, apricot, banana, blueberry, cherry, cranberry, Michelia champaca, Jasminum
grape, jasmin, lily, melon, pineapple, raspberry, violet

Methyl benzoate Herb, lettuce, prune, violet, cherry, clove, cranberry, Michelia champaca, Jasminum sambac, Cananga odorata,
jasmin, mint, pineapple, raspberry, strawberry, vanilla Ribes nigrum

Butyl benzoate Banana, cherry, geranium, orchid, papaya, pineapple, Michelia champaca, Jasminum and strawberry
rose, strawberry

Anisyl acetate Juniper berry, raspberry, vanilla, fig, coconut, cherry, Cananga odorata, Clavija
balsam, plum, euerganea, Jacquinia keyensis, Jacquinia sprucei

Anisyl propionate Sweet, fruity, floral, vanilla Pimpinella anisum L.

2-phenethyl acetate Apricot, peach vanilla, tutti-frutti Freesia magnolia, Hyacinth reseda

2-phenethyl hexanoate Pineapple, rose, apple, banana, fruity Michelia champaca

1-phenethyl acetate Rose, honey, fruity, floral Michelia champaca, Eschweilera coriacea

p-cresyl acetate Apple blossom, cherry blossom Prangos uechtritzii, Cananga odorata

Cinnamyl acetate Cinnamon, oriental, rose, apricot, guava Psidium guajava, Laurus nobilis, Cinnamomum verum

Cinnamyl propionate Apricot, almond, apple, banana, coconut, grape, cherry, Cinnamon, Michelia sp., Prunus sp., Tanacetum sp. and
blackberry, vanilla, strawberry, plum melon

Eugenyl acetate Clove, cinnamom Syzygium aromaticum, Cinnamomum verum


98  et al. / Trends in Food Science & Technology 69 (2017) 95e105
A.G.A. SA

Table 2
Synthesis of benzyl esters with benzyl alcohol as acyl acceptor.

Aromatic Ester Acyl Donor Molar Ratio Enzyme Solvent Temperature Time Conversion References
Acyl Acceptor:
Acyl Donor

Benzyl acetate Vinyl acetate 2: 3.5 Immobilized Pseudomonas Heptane 50  C 4h 99% (Badgujar et al., 2015)
fluorescens lipase, 36 mg

Benzyl acetate Vinyl acetate 1: 1 Pseudomonas aeruginosa Heptane 50 C 3h 88.8 ± 0.2% (Singh et al., 2008)
lipase, 50 mg
Benzyl propionate Vinyl propionate 2: 5 Immobilized Pseudomonas Isooctane 50  C 2.5 h 99% (Badgujar & Bhanage, 2014b)
cepacia lipase, 54 mg
Benzyl butyrate Vinyl butyrate 1: 2.7 Immobilized Pseudomonas Isooctane 52  C 3h 99% with (Badgujar & Bhanage, 2015)
cepacia lipase, 300 mg ultrasound
technique
Benzyl butyrate Butiric acid 1: 1 Novozym 435®, 200 mg Methyl tert-butyl 52  C 24 h 82% (Jeromin & Zoor, 2008)
ether

Benzyl laurate Lauric acid 5: 1 Aspergillus oryzae lipase Solvent-free 30 C 6h 80% (Shintre et al., 2002)
Benzyl oleate Oleic acid 1: 1 Novozym 435®, 12.5 mg Solvent-free 80 
C 1h 94% (Vosmann et al., 2008)
Benzyl cinnamate Cinnamic acid 3: 1 Lipozyme TL-IM®, 30 mg Isooctane 40 
C 24 h 97.3% (Wang et al., 2015)
Benzyl cinnamate Cinnamic acid 2.6: 1 Lipozyme TL-IM®, 31 mg.mL1 Isooctane 40 
C 27 h 97.7% (Zhang et al., 2016)

Table 3
Synthesis of cresyl and anisyl esters with cresol and anisyl alcohol as acyl acceptor, respectively.

Aromatic Ester Acyl Donor Molar Ratio Enzyme Solvent Temperature Time Conversion References
Acyl Acceptor:
Acyl Donor

p-cresyl acetate Vinyl acetate 2: 3.5 Immobilized Pseudomonas Heptane 50  C 12 h 69% (Badgujar et al., 2015)
fluorescens lipase, 36 mg
p-cresyl acetate Vinyl acetate 1: 5 Steapsin lipase, 70 mg Hexane 55  C 60 h 81.0 ± 1.3% (Dhake et al., 2012)
p-cresyl propionate Vinyl propionate 1: 2 Immobilized Pseudomonas Heptane 40  C 1.5 h 99% (Badgujar et al., 2016)
cepacia lipase, 20 mg
o-cresyl butyrate Vinyl butyrate 1: 2.7 Immobilized Pseudomonas Isooctane 52  C 3.5 h 99% with ultrasound (Badgujar & Bhanage, 2015)
cepacia lipase, 300 mg technique

Anisyl acetate Vinyl acetate 2: 3.5 Immobilized Pseudomonas Heptane 50 C 4.5 h 99% (Badgujar et al., 2015)
fluorescens lipase, 36 mg
Anisyl acetate Vinyl acetate 1: 5 Steapsin lipase, 70 mg Hexane 55  C 48 h 99.0 ± 0.3% (Dhake et al., 2012)
Anisyl acetate Vinyl acetate 1: 5 Immobilized Rhizopus oryzae Hexane 45  C 48 h 99% (Dhake et al., 2011)
lipase, 50 mg
Anisyl butyrate Vinyl butyrate 1: 2.7 Immobilized Pseudomonas Isooctane 52  C 3h 99% with ultrasound (Badgujar & Bhanage, 2015)
cepacia lipase, 300 mg technique

Anisyl propionate Vinyl propionate 1: 2 Immobilized Pseudomonas Heptane 40 C 1.5 h 99% (Badgujar et al., 2016)
cepacia lipase, 20 mg

cinnamyl, phenethyl, benzoate and cinnamate esters, respectively. in 24 h (Jeromin & Zoor, 2008).
Aromatic esters production via enzymatic route is a green pro- Transesterification is an alternative to avoid the drawbacks
cess alternative and the process variables are important for the final found in the esterification and improve the ester conversion and
conversion and reaction yield. The molar ratio of substrates, type yield. The transesterification occurs between alcohols (acyl
and amount of enzyme, nature of substrates, temperature, reaction acceptor) and esters (acyl donor). Vinyl esters are usually employed
time, agitation speed and use of organic solvents have a great in the transesterification and thanks to the isomerization of the
impact on the process production. The optimization of the process vinyl alcohol, that prevents the opposite reaction, it is able to
by the study of the individual effects and interactions is useful to promote higher conversions in the synthesis process. However, the
achieve high product yield and conversion (Geng, Wang, Qi, & Su, co-product acetaldehyde may have unfavorable deactivation effect
2012; Paroul, Grzegozeski, Chiaradia, & Treichel, 2012; Stencel & on some enzymes, so another alkyl esters can be used in order to
Leadbeater, 2014). overcome this drawback, such as ethyl acetate and ethyl butyrate
(Badgujar & Bhanage, 2015; Kuo, Chiang, Ju, & Chen, 2012; Kuo
4.1. Effect of acyl donors et al., 2014).
Some researchers have shown this behavior related for esteri-
The main reaction for the esters production is the esterification, fication and transesterification in the aromatic ester production,
which occurs between the alcohols (acyl acceptor) and acids (acyl like the synthesis of benzyl propionate with propionic acid as acyl
donor). A disadvantage of the enzymatic esterification is the pos- donor. The results showed lower initial reaction rate and conver-
sibility of low yield production, because of a severe enzyme activity sion (8% in 2.5 h) for the enzymatic catalysis, probably due to the
deactivation due to the high acid concentration in the reaction acting of acid as a potent inhibitor of enzyme activity. On the other
medium (Badgujar & Bhanage, 2014b). Therefore, the choice of the hand, tests made with methyl propionate and vinyl propionate as
substrates can affect directly the conversion and yield results. acyl donor indicated that the reaction with vinyl propionate had the
Meanwhile, researchers have presented that it is possible to reach conversion increased substantially (99%) than that with methyl
good results using acid and alcohol for ester synthesis, as shown in propionate (17%) in the same conditions. The low conversion
a study of esterification between benzyl alcohol and butyric acid reached when methyl propionate was used as acyl donor was
(molar ratio 1:1) to produce benzyl butyrate, reaching 82% of yield probably due to the presence of methanol that is a side product of
 et al. / Trends in Food Science & Technology 69 (2017) 95e105
A.G.A. SA 99

Table 4
Synthesis of eugenyl and cinnamyl esters with eugenol and cinnamyl alcohol as acyl acceptor, respectively.

Aromatic Ester Acyl Donor Molar Ratio Enzyme Solvent Temperature Time Conversion References
Acyl Acceptor:
Acyl Donor

Eugenyl acetate Acetic anhydride 1: 5 Novozym 435®, 1 wt % Supercritical CO2, 40  C 1h 33.2 ± 0.3% (Santos et al., 2016)
10 MPa
®
Eugenyl acetate Acetic anhydride 1: 5 Lipozyme 435 , 10 wt % Supercritical CO2, 40  C 1h 20.3 ± 1.0% (Santos et al., 2016)
10 MPa
®
Eugenyl acetate Acetic anhydride 1: 5 Lipozyme TL-IM , Solvent-free 70  C 2h 94.3% (M. J. A. Silva et al., 2015)
5 wt %
Eugenyl acetate Acetic anhydride 1: 5 Novozym 435®, 10 wt % Solvent-free 60  C 6h 99.9% (Vanin et al., 2014)
Eugenyl acetate Acetic anhydride 1: 3 Novozym 435®, 5.5 wt % Solvent-free 50  C 6h 99.86% (Chiaradia et al., 2012)
Eugenyl benzoate Benzoic acid 4: 1 Immobilized Rhizomucor Chloroform 60  C 6h 56.1% (Manan et al., 2016)
miehei lipase, 15 mg
Eugenyl benzoate Benzoic acid 1: 1.22 Immobilized Staphylococcus Chloroform 41  C 6h 75% (Horchani et al., 2010)
aureus lipase, 240 UI
Eugenyl caprylate Caprylic acid 1.13: 1 Novozym 435® and Hexane 56.8  C 4h > 80% (Radzi et al., 2016)
Lipozyme TL-IM®
Eugenyl caprylate Caprylic acid 2: 1 Lipozyme TL-IM®, Solvent-free 65  C 4.3 h 72% (Chaibakhsh et al., 2012)
100 mg
Cinnamyl acetate Vinyl acetate 1: 2 Esterase, 10 g.L1 Hexane 
40 C 12 h 99% (Gao et al., 2016)
Cinnamyl acetate Vinyl acetate 1: 2 Novozym 435®, 0.2% Solvent-free 40  C 20 min 99.99% (Tomke & Rathod, 2015)
Cinnamyl acetate Vinyl acetate 1: 5 Immobilized Rhizopus oryzae Hexane 45  C 24 h 99% (Dhake et al., 2011)
lipase, 50 mg
Cinnamyl acetate Ethyl acetate 1: 15 Novozym 435®, 2.67 g.L1 Solvent-free 40 C 3h 90% (Geng et al., 2012)
Cinnamyl acetate Acetic acid 1: 2 Immobilized procine Hexane 35  C 10 h 62.6% (Wu et al., 2014)
pancreatic lipase, 3 g
Cinnamyl acetate Vinyl acetate 1: 2 Novozym 435®, 10 mg Toluene 40  C 1h 96% (Yadav & Devendran, 2012)
Cinnamyl propionate Vinyl propionate 1: 2 Immobilized Pseudomonas Heptane 40  C 1.5 h 99% (Badgujar et al., 2016)
cepacia lipase, 20 mg
Cinnamyl laurate Lauric acid 1: 1 Novozym 435®, 50 mg Toluene 30  C 2h 60% (Yadav & Dhoot, 2009)

Table 5
Synthesis of phenethyl esters with phenethyl alcohol as acyl acceptor.

Aromatic Ester Acyl Donor Molar Ratio Enzyme Solvent Temperature Time Conversion References
Acyl Acceptor:
Acyl Donor

1-phenethyl acetate Vinyl acetate 0.4: 1.1 Novozym 435®, 10 mg Liquid CO2, 20  C 1h 32% (Hoang & Matsuda, 2016)
7 MPa
2-phenethyl acetate Vinyl acetate 2: 3.5 Immobilized Pseudomonas Heptane 50  C 4h 99% (Badgujar et al., 2015)
fluorescens lipase, 36 mg
2-phenethyl acetate Vinyl acetate 1: 3.65 Immobilized Candida Hexane 35.85  C 38.8 h 95.3 ± 2.6% (Kuo et al., 2014)
rugosa lipase
2-phenethyl acetate Vinyl acetate 1: 1 Novozym 435®, 122.5 mg Hexane 57.8  C 79 min 85.4 ± 0.4% (Kuo et al., 2012)
1-phenethyl acetate Vinyl acetate 0.5: 2 Steapsin lipase, 70 mg Hexane 55  C 24 h 48.0 ± 0.5% (Dhake et al., 2012)
2-phenethyl propionate Vinyl propionate 1: 2 Immobilized Pseudomonas Heptane 40  C 1.5 h 98% (Badgujar et al., 2016)
cepacia lipase, 20 mg
2-phenethyl butyrate Butter oila 3: 1 Lipozyme TL-IM®, 20 wt% e 40  C 8h 50.8 ± 1.9% (Li et al., 2014)
2-phenethyl hexanoate Butter oila 3: 1 Lipozyme TL-IM®, 20 wt% e 40  C 8h 66.6 ± 2.7% (Li et al., 2014)
2-phenethyl caffeate Caffeic acid 1:71 Novozym 435®, 2938 PLU Isooctane 70  C 9.6 h 93.1 ± 0.4% (Chen et al., 2011)
with ultrassound
technique
a
Fatty acids from butter oil: butyric 10%; caproic 5%, caprylic 2.6%; capric 5%; lauric 5%; myristic 12%; palmitic 27%; stearic 10%; and oleic 23%.

Table 6
Synthesis of benzoate and cinnamate esters with benzoic acid and cinnamic acid as acyl donor, respectively.

Aromatic Ester Acyl Acceptor Molar Ratio Enzyme Solvent Temperature Time Conversion References
Acyl Acceptor:
Acyl Donor

Methyl benzoate Methanol 12: 7 Lecitase Ultra™, Dichloromethane 40  C Continuous flow 41% (Gumel & Annuar, 2016)
123 U.g1 rate 0.1 mL/min
Methyl benzoate Methanol 6.2: 5 Candida rugosa lipase, Hexane and 37  C 75 h 100% (Leszczak & Tran-Minh, 1998)
10 mg.mL1 Toluene
Hexyl benzoate Hexanol 19: 1 Immobilized Bacillus Solvent-free 40  C 2.7 d 24% (Krause, Hilterhaus, Fieg, &
subtilis esterase, 6 wt % Liese, 2009)
Heptyl benzoate N-heptanol 1: 1 Novozym 435®, 50 mg Cyclohexane 60 
C 24 h 100% (Giunta et al., 2015)
Benzyl benzoate Benzyl alcohol 2: 1 Novozym 435®, 300 mg Solvent-free 55 
C 100 h 90% (Gryglewicz et al., 2000)
Ethyl cinnamate Ethanol 3: 1 Lipozyme TL-IM®, 30 mg Isooctane 50 
C 24 h 99% (Wang et al., 2016)
Butyl cinnamate Butanol 15: 1 Novozym 435®, Isooctane 55 
C 72 h 60.7% (Jakoveti
c, Jugovi
c, et al., 2013)
3 wt%
Oleyl cinnamate Oleyl alcohol 6: 1 Novozym 435®, 20 mg Isooctane and 55  C 12 d 100% (Lue et al., 2005)
2-butanone
® 
Geranyl cinnamate Geraniol 2: 1 Novozym 435 , 60 mg Hexane 65 C 15 min 88 ± 2% (Shinde & Yadav, 2015)
100  et al. / Trends in Food Science & Technology 69 (2017) 95e105
A.G.A. SA

the transesterification reaction and was able to compete with complex formation, which resulted in a higher conversion, but a
benzyl alcohol for nucleophilic attack on carbonyl ester, inhibiting very high quantity of the enzyme leads to their agglomeration,
the reaction rate and reducing the conversion (Badgujar & Bhanage, which can block the substrate sites for enzyme attack (Badgujar
2014b). Other study presented the benzyl butyrate synthesis using et al., 2016; Khan & Rathod, 2015; Wang et al., 2015). Thereby,
butyric acid, ethyl butyrate and vinyl butyrate for esterification and the excess of the enzyme amount has no contribution to the re-
transesterification, respectively. The results showed that the reac- action rate and must be controlled in order to guarantee high
tion with vinyl butyrate had a better conversion (~99%) than that conversions and low cost.
with butyric acid (~40%) and ethyl butyrate (~50%) in similar re-
action parameters (Badgujar & Bhanage, 2015). 4.3. Effect of molar ratio

4.2. Effect of enzyme The molar ratio is the ratio of acyl donor (i.e. acids, esters or
anhydrides) and acyl acceptor (i.e. alcohols) used in the ester
The utilization of immobilized enzymes in the synthesis of biosynthesis. The quantity of the substrates is an important factor
many products aims high process specificity, selectivity, pro- to determine their influence on enzyme activity and subsequent
ductivity and easy recuperation. Several aspects might influence reaction rate (Badgujar et al., 2015). Thereby, the study of the best
the activities of these biocatalysts, such as the enzyme source and molar ratio is a very important parameter for any catalysis system
the nature of immobilization support. As the main source, mi- that aims to reach great conversions, no loss of the enzyme activity
croorganisms can produce many enzymes and naturally present and no waste of reagents.
strong influence in the final structural characteristics, which The acid, at high concentrations, may present an inhibitory ef-
modify the biocatalysts properties and activities even in similar fect and cause a reduction in the enzyme catalytic activity
reaction parameters (Martins, Da Silva, Schein, & Garcia-Galan, (Badgujar & Bhanage, 2014b, 2015; Badgujar et al., 2016). In order to
2014). counterbalance the inhibitory influence of the acid in the enzyme
Moreover, the nature of the immobilization support can alter activity on the esterification, some researchers recommend the use
the lipases properties making difficult the access of the substrate to of a higher concentration of alcohol. However, a large increase of
the enzyme active site. The impediment to access the active site alcohol amount may also decrease the conversion due to the polar
may occur due to the hydrophilic or hydrophobic characteristic of character of this reactant, which shows hydrophilic interaction
the support material, and can cause partition of the substrates and with the water layer present on the enzyme surface and causes
products or blocking of the active site during the immobilization changes in the protein structure of the enzyme with consequent
process. The immobilization protocol is also an important aspect inhibition and reduction of the activity (Badgujar & Bhanage, 2015;
because it changes the enzyme activities even when it is immobi- Gumel & Annuar, 2016; Shinde & Yadav, 2014; Wang et al., 2015).
lized on the same support (Martins et al., 2014). The high quantity of alcohol in the formulation also leads to
Many commercial lipases are available for purchased and widely higher costs, from the economic point of view, due to the need of
used in a range of catalysis. The commercial immobilized lipases purification. Therefore, researchers have been studying different
most used are Novozym 435®, Lipozyme TL-IM® and Lipozyme RM- substrates molar ratio to avoid unnecessary amounts of alcohol. The
IM®, from Novozymes®, and each one has a different lipase source, production of benzyl acetate (Majumder & Gupta, 2010) and ethyl
support material and immobilization method. Novozym 435® is the cinnamate (C. K. Sharma, Chauhan, & Kanwar, 2011) with 1:1 M
Candida antarctica B lipase, immobilized in Lewatit VP OC 1600, a ratio (alcohol/acid) reaches 82% and 54.1% of conversion, respec-
macro-porous resin with a hydrophobic surface. Lipozyme TL-IM® tively, showing the possibility to reach good conversion results
is a Thermomyces laguginosus lipase, immobilized in a hydrophilic with no need of a higher concentration of alcohol.
gel silicate and Lipozyme RM-IM® is a Rhizomucor miehei lipase, Another study showed the effect of the substrates molar ratio on
immobilized in a support of Duolite ES562, a weak anion-exchange the enzyme activity in the biocatalysis of benzyl propionate. Results
resin based on phenol-formaldehyde copolymers. Regarding the indicated a conversion decrease with the increase of benzyl alcohol
immobilization protocols, Novozym 435® is prepared via interfacial quantity. This behavior present the inhibitory effect of the benzyl
activation on the hydrophobic surface of the support, while Lip- alcohol, which may form a dead-end inhibition complex with the
ozyme RM-IM® and Lipozyme TL-IM® are immobilized via anionic biocatalyst at higher concentrations, reducing the enzymes binding
exchange (Martins et al., 2014). All these differences of the bio- sites and consequently decreases the reaction rate (Badgujar &
catalysts make the comprehension of the substrates interactions on Bhanage, 2014a).
the enzymes activities in different systems more complex. Some researchers investigated the effect of the molar ratio in
In laboratory scale, studies easily show how these influences (i.e. the conversion of the subtrates for the eugenyl acetate and phe-
lipase source, immobilization method and type of immobilization nylethyl acetate synthesis. For eugenyl acetate production, using
support) affect the reaction conversion results. Researchers evalu- various molar ratios (1:1, 1:3, 1:5 eugenol/acetic anhydride), the
ated the efficacy of Novozym 435®, Lipozyme RM-IM® and Lip- maximum yield was 99.86% after 6 h of reaction at 50  C with
ozyme TL-IM® to synthesize cinnamyl laurate in similar conditions. 1:3 M ratio (Chiaradia, Paroul, Cansian, & Júnior, 2012). The 2-
The final conversion of 60, 16.5 and 9% in 2 h, respectively, showing phenylethyl acetate synthesis, with two different molar ratios of
the Novozym 435® is the most adequate immobilized enzyme to substrates (2:1 and 4:1 vinyl acetate/2-phenylethyl alcohol), pre-
this reaction system. These results indicate that the different lipase sented the best conversion of 94.81% at 36 h with 4:1 M ratio (Kuo
sources and immobilization material can really affect the conver- et al., 2014).
sion results (Yadav & Dhoot, 2009).
In addition, another important factor is the enzyme amount, 4.4. Effect of reaction media
which regards some attention for economic and industrial
viability. In industrial applications, there is a great interest in the Esterification is generally a water-limited reaction because the
study of the process parameters to achieve lower production costs equilibrium by hydrolytic enzymes is in favor of the hydrolysis
and to obtain the maximum productivity. Enzyme amount affects (inverse reaction). The esterification resulted in the formation of
significantly the economic feasibility of the entire process. The water as a reaction by-product and its removal using molecular
higher biocatalyst amount assisted rapid enzyme-substrate sieves might enhance the synthesis of the ester by pushing the
 et al. / Trends in Food Science & Technology 69 (2017) 95e105
A.G.A. SA 101

reaction equilibrium in the forward direction (C. K. Sharma et al., 4.5. Effect of temperature
2011).
The use of organic solvent media for enzymatic aromatic ester Temperature is a crucial key parameter for biocatalysis, which
synthesis presents some advantages, such as increased substrates facilitates reactants solubility in reaction media, reduces mixture
and products solubility and the shifting of the thermodynamic re- viscosity, boosts the molecular collision interface, reduces mass
action equilibrium to favor esterification over hydrolysis (Akacha & transfer limitations and assists the interactions between enzyme
Gargouri, 2015; Patel, Gajera, Gupta, & Manocha, 2015). The nature particles and substrates. Temperature increases the molecular
of the solvent influences the activity, selectivity and stability of the collision interaction and causes a decrease in the energy barrier
enzymes. In general, lipases are more stable when suspended in between reacting molecules and enzyme-substrate complex for-
non-polar solvents that have low solubility in water. The choice of mation, which causes improvement in initial reaction rate and
the organic solvent for the enzymatic reactions is essential to offer conversion. Enzymes are biocatalysts, stables at an optimal range of
good solubility of the substrate in reaction media, without affecting temperature, however, some modification on the catalytic activity
the catalytic power of the enzyme (Badgujar et al., 2016; Shinde & and stability can occur when the temperature increases beyond the
Yadav, 2014). optimum temperature, leading to enzyme thermal deactivation
The log P concept (the partition coefficients between water and (Badgujar et al., 2015; Ceni, Lerin, Conto, & Brancher, 2010; Gumel
solvent, which correlated with solvent hydrophobicity) determines & Annuar, 2016; Manan et al., 2016).
the effect of the organic solvents on the reaction rate and the yield. In aromatic ester production, many researchers showed that the
Solvents with low log P values are more hydrophilic and tend to optimum temperature of an enzyme might change depending on
strip away the water present on the surface of the enzyme. Solvents the system used. The utilization of Novozym 435® in the synthesis
with intermediate log P (>4) are more suitable for the aromatic of 2-phenethyl caffeate and cinnamyl acetate with different tem-
esters synthesis (Singh, Singh, Singh, & Chisti, 2008; Wang, Zhang, peratures (70 and 40  C, respectively) showed that the large range
Zhang, & Chen, 2016; Wang et al., 2015; Yadav & Devendran, 2012). of temperature did not present great effect in the final conversion
Some studies have shown good results by the use of isooctane for for both esters, which remained 93.1 and 96% for 2-phenethyl
benzyl propionate, benzyl butyrate, benzyl cinnamate, anisyl caffeate and cinnamyl acetate, respectively (Chen, Chen, Chang, &
butyrate and oleyl cinnamate (Badgujar & Bhanage, 2014b, 2015; Shieh, 2011; Yadav & Devendran, 2012).
Lue, Karboune, Yeboah, & Kermasha, 2005; Wang et al., 2015;
Zhang, Zhang, Che, & Wang, 2016), heptane for 3-phenylpropyl 5. Ultrasound-assisted synthesis technology
acetate, 2-methylbenzyl acetate, phenyl acetate and hexyl benzo-
ate (Badgujar et al., 2015; Shinde & Yadav, 2014), hexane for benzyl Ultrasound is an emergent energy-efficient technique and
acetate, 2-methylbenzyl acetate, 2-phenoxybenzyl acetate and p- recently has been used for flavor and aromatic ester synthesis. The
cresyl acetate (Dhake et al., 2012, 2011). technique is able to increase the initial rate and enhance mass
On the other hand, the use of solvents may have some draw- transfer of the reactions. Furthermore, ultrasound method is a
backs associated with the separation costs and presence of harmful green technology with high efficiency, economic performance and
residual substances at the final product, which could be prejudicial low instrumental requirement (Khan & Rathod, 2015; Paludo et al.,
to human health. In this scenario, lipase-mediated synthesis of 2015; Tomke & Rathod, 2015; Zheng, Wang, Huang, & Guo, 2013).
aromatic esters under solvent-free systems has significant impor- Ultrasound is the sound energy at frequencies above the range
tance due to the absence of toxic solvents, which eliminates the that is audible to human beings (>16 kHz). The cycles of
need of recuperation, downstream and purification processes and compression and rarefaction of the sound waves can generate a
reduces the environment hazards (Garlapati & Banerjee, 2013; phenomenon known as cavitation, which comprises the formation,
Geng et al., 2012; Paroul et al., 2012; M. J. A.; Silva et al., 2015). enlargement and collapse of bubbles, increasing the rate of enzy-
The most important advantage is the possibility of high reaction matic reactions. When cavitation bubbles collapse near the phase
conversions, as shown in benzyl acetate synthesis study under a boundary of two immiscible liquids, it can provide a very efficient
solvent-free system with final conversion of 100% (Majumder, stirring. The collapse of the bubbles produces localized supercritical
Singh, Dutta, & Sadhukhan, 2006). conditions (high temperature and high pressure). However, ultra-
Recently, there is an increase of interest in the use of green sound reactions at very high intensity can lead to the disruption of
solvents for enzymatic ester production, such as dense gases like the enzyme structure and low intensity of ultrasound will not result
supercritical carbon dioxide, due to the low cost, non-toxicity, non- in the desirable cavitation effect (Bansode & Rathod, 2014; Ceni,
flammability, inertness, recyclable, environmentally friendly and Silva, Lerin, & Oliveira, 2011; Chen et al., 2011). Researchers pro-
moderate critical properties (Pc ¼ 7.38 MPa and Tc ¼ 304.2 K). duced benzyl butyrate, anisyl butyrate and o-cresyl butyrate with
Supercritical fluids are defined as fluids above their critical tem- ultrasound technique and reached 99% of yield, in 3 h at 52  C, for
perature and pressure, having liquid-like densities and gas-like these aromatic esters. On the other hand, the reactions performed
diffusivities. Therefore, they appear as suitable solvents for enzy- without the use of ultrasound at the same conditions presented low
matic reactions. The low viscosity and high diffusivity of super- conversions of 57%, 55% and 43% for benzyl butyrate, anisyl buty-
critical CO2 also serve to provide favorable mass transfer properties rate and o-cresyl butyrate, respectively (Badgujar & Bhanage, 2015).
(Ceni, Silva, Lerin, & Charin, 2010; Longo & Sanrom an, 2006; Santos These results show the efficiency of the utilization the ultrasound
et al., 2016). There are a few studies using supercritical fluids (CO2, technique in aromatic esters production, being a promising tech-
10 MPa) for aromatic esters synthesis, such as benzyl acetate nology to increase the initial rate of reaction and yields.
(Tewari, Ihara, Phinney, & Mayhew, 2004) and eugenyl acetate
(Santos et al., 2016). The experimental apparatus used for the 6. System optimization
synthesis of eugenyl acetate is a high-pressure stirred-bath reactor
unit, using Novozym 435® as the biocatalyst and eugenol and acetic Optimization plays a significant role in the commercial success
anhydride as substrates. After 1 h of reaction under CO2 pressurized of the biotechnological industry based on quality, cost and the
the samples reached 33.2% of conversion (Santos et al., 2016). process performance. The conventional method of optimization
However, there is a lack of investigation around the use of super- requires screening of a large number of variables, many experi-
critical fluids for aromatic ester synthesis. ments, with plenty time and resources. Experimental design
102  et al. / Trends in Food Science & Technology 69 (2017) 95e105
A.G.A. SA

approach provides an easy and efficient evaluation of the main the lipase to combine with ethyl acetate (Geng et al., 2012).
reaction variables, such as temperature, time reaction, enzyme The Lineweaver-Burk graph can determine the inhibition from
amount and the molar ratio of substrates, which improves the substrates, by plotting initial rates at different concentration of
biocatalyst activity and the conversion in the esters synthesis substrates (Badgujar et al., 2016). In the study of the benzyl pro-
(Chaibakhsh, Basri, Anuar, & Rahman, 2012; Narwal et al., 2016; pionate synthesis using vinyl propionate as acyl donor, Lineweaver-
Shinde & Yadav, 2014, 2015). Burk plot showed a decrease in the initial rate with a high con-
The response surface methodology (RSM) technique is an centration of benzyl alcohol, which reveals that the alcohol acts as
innovative powerful tool for the aromatic esters synthesis, able to an inhibitor of the enzyme for the benzyl propionate synthesis
determine the optimum reaction conditions necessary to scale up (Badgujar & Bhanage, 2014b).
the process and to reduce the number and cost of experimental In the ternary complex mechanism (order Bi-Bi), the lipase (E)
tests need to provide statistically acceptable results (Radzi, Hanif, & binds with the acyl donor (Ad) to form enzyme-acyl donor complex
Syamsul, 2016; Zhang et al., 2016). The statistical method of RSM (E-Ad). Then, the acyl acceptor (Ac) combines with E-Ad to form a
was employed to optimized the synthesis of eugenyl benzoate, by ternary complex (E-Ad-Aa). This ternary complex isomerizes to
5-level-4-factor central composite design (CCD) with the parame- another ternary complex (E-Q-W), which releases the desired
ters of incubation time (2e18 h), temperature (30e70  C), molar product (Q) and water (W) and frees the enzyme (Badgujar et al.,
ratio of acid to alcohol (1:1e1:5) and enzyme loading (5e45 mg). 2016; Yadav & Dhoot, 2009). Some studies describes the ternary
The highest yield of eugenol benzoate was 56.1%, at 60  C, 6 h of complex mechanism for the enzymatic synthesis of aromatic esters,
reaction, 15 mg of enzyme loading and molar ratio 1:4 (Manan such as cinnamyl laurate using cinnamyl alcohol (Ac) and lauric
et al., 2016). The same method was used for the synthesis of acid (Ad) (Yadav & Dhoot, 2009). Other research showed the kinetic
benzyl cinnamate; the reaction variables were time (18e30 h), parameters calculated for the benzyl propionate production, sug-
temperature (30e50  C), molar ratio of acid to alcohol (1:1e1:5) gesting that the ternary complex mechanism fits the data (Badgujar
and enzyme loading (20e40 mg). Under optimized conditions & Bhanage, 2014b). The same mechanism explains the cinnamyl
(40  C, 31 mg of enzyme loading, 1:2.6 M ratio and 27 h) the yield propionate synthesis with cinnamyl alcohol and vinyl propionate;
reaches 97.7% (Zhang et al., 2016). however, with a high concentration of cinnamyl alcohol, there is
the formation of a dead-end binary inhibition complex between the
enzyme and the cinnamyl alcohol, instead of the enzyme and vinyl
7. Enzymatic kinetics propionate, reducing the reaction rate and conversion (Badgujar
et al., 2016).
Kinetic modeling and mechanistic study of a reaction are
important aspects for reactor designing and scale up. Several 8. Biological activities of flavor esters
mechanisms are available to explain lipase-catalyzed reactions and
usually follows Ping-Pong Bi-Bi or ternary complex (order Bi-Bi) Emerging studies demonstrate that aromatic esters may present
mechanism (Kuo et al., 2014; Yadav & Dhoot, 2009). biological activities, such as antioxidant, antimicrobial and larvi-
In the Ping-Pong Bi-Bi mechanism, the reaction occurs between cidal properties (Berger, 2009). Studies have shown that benzyl and
two substrates and a product is released. For the aromatic ester methyl benzoate, benzyl and cinnamyl acetate have larvicidal
synthesis, firstly the lipase (E) combines with the acyl donor (Ad) properties against larvae of Aedes aegypti mosquitoes (Pavela,
(i.e. acid, ester or anhydride) and forms an enzyme-acyl donor 2015). Eugenyl acetate, an ester derived from eugenol, the main
complex (E-Ad). E-Ad is transformed into an intermediate complex compound of clove essential oil, also had antimicrobial and larvi-
(E-Ac) by molecular isomerization. Then, the acyl acceptor (Aa) (i.e. cidal properties (Chiaradia et al., 2012; M. J. A.; Silva et al., 2015).
alcohol) combines with E-Ac to form an enzyme-acyl-alcohol The phenolic compounds (e.g. flavonoids) present advantageous
complex (E-Ac-Aa), which isomerizes into another enzyme- biological and physiological properties, such as antimicrobial, pest
product ester complex (E-Q) and releases the desired product (Q) repellents, anti-allergenic, anti-inflammatory, anticarcinogenic and
and frees the enzyme. Equation (1) defines the reaction rate (V) of antimutagenic properties. The modification of these compounds,
the Ping-Pong Bi-Bi mechanism (Geng et al., 2012; Jakoveti c, via esterification to produce useful derivatives with a great com-
Jugovi c, Gvozdenovic, & Bezbradica, 2013). mercial importance, has been the subject of increased interest
aiming to improve the solubility and stability of these compounds
Vmax :½Ad0 :½Aa0 (Chen et al., 2011; Lue et al., 2005; Vosmann, Wiege, Weitkamp, &
V ¼ (1)
KAa :½Ad0 þ KAd :½Aa0 þ ½Ad0 :½Aa0 Weber, 2008; Yadav & Dhoot, 2009; Zheng et al., 2013).
Benzyl benzoate is one of the oldest drugs used for the treat-
Sometimes the acyl acceptor (Aa) acted as a competitive in-
hibitor of lipase, forming a dead-end complex (E-Aa), which ment of scabies, a highly contagious skin infection. It is common
worldwide, but is more conspicuous in the areas with poor sani-
impeded the lipase to combine with the acyl donor (Ad). An addi-
tional parameter is necessary to describe the reaction rate with tation and overcrowding. The mite Sarcoptes scabiei burrows into
the skin and consumes the epidermis, resulting in inflammation,
inhibition and Equation (2) describes adequately the kinetic
expression (Geng et al., 2012; Jakovetic, Jugovi
c, et al., 2013). allergy reactions and pruritic lesions. There are adversities associ-
ated with the use of benzyl benzoate, like severe burning sensation,
Vmax :½Ad0 :½Aa0 itching and scaling of the skin, after repeated use. Therefore, the
V ¼   (2) topical use of the benzyl benzoate is not recommended and a
½Aa
KAa :½Ad0 þ KAd :½Aa0 1 þ KI;Aa0 þ ½Ad0 :½Aa0 suitable carrier system can be a good alternative to protect the skin
against the adverse effects. Thereby, the encapsulation technique
The Ping-Pong Bi-Bi mechanism describes the enzymatic syn- can revitalize the usage of this compound (G. Sharma, Dhankar,
thesis of some aromatic esters, such as cinnamyl acetate with ethyl Thakur, & Raza, 2015).
acetate (Ad) and cinnamyl alcohol (Ac). The increase of cinnamyl
alcohol concentration decelerated the initial rate of the reaction 9. Encapsulation of flavors
and shows that cinnamyl alcohol acted as a competitive inhibitor of
the lipase due to the formation of a dead-end complex, which stops Flavor encapsulation can be a useful tool to promote product
 et al. / Trends in Food Science & Technology 69 (2017) 95e105
A.G.A. SA 103

functionality and stability. Encapsulation describes the use of This review paper presents the current use of biocatalysis for the
different processes where active ingredients are stored in a sec- aromatic esters production by esterification and transesterification
ondary material to include some level of protection against vola- studied in the academic area. There are many prospects for
tilization and degradation. The benefits of the flavor encapsulation expansion the biotechnological route to the industry in the future.
are the ease of handling, stability improvement, texture creation, However, more investigation would be helpful to understand better
adjustable properties (particle size) and the controlled release of the potential of ultrasound, the optimization process, reaction ki-
the aroma (Rodríguez, Martín, Ruiz, & Clares, 2016). Many materials netics and scale up for well assessment of economic and industrial
are suitable as matrix for ester compounds encapsulation and the viability in aromatic ester synthesis.
main requisite is the film forming property that is usually related to
materials derived from natural or synthetic sources like polymers 11. Conclusions
and lipids (Zhu, Xiao, Zhou, & Yi, 2012). The encapsulation effi-
ciency is a delicate balance between stability over the product The production of aromatic esters by the lipase-catalyzed re-
lifetime and the desired release. The final use of the product, action is showing to be a great alternative with several advantages
technology costs, scale up and legal aspects to choose the best over production via chemical route. The study of the process pa-
system are important aspects that must be evaluated (Rodríguez rameters (type of substrates, molar ratio, temperature, type and
et al., 2016). amount of enzyme and reaction media) and their interaction are
Many techniques are available to encapsulate diverse bioac- very important. The system optimization is a great tool to achieve
tive compounds, like drugs, phenols, flavor, colorants, lipids, the best reaction conditions with maximum yield. The knowledge
enzymes, vitamins and minerals. Spray drying is the most used of reaction kinetics and mechanism is extremely important for
technique for encapsulation of food ingredients in the industry lipase-catalyzed reactions to scale up and usually follows Ping-
due to the economical, flexible and continuous operation. Mi- Pong Bi-Bi or ternary complex (order Bi-Bi) mechanism. In addi-
croparticles formed by this technique are stables, good for food tion to its fragrance, aromatic esters can present some biological
additives and flavors applications (Fang & Bhandari, 2010). activities, further increasing the interest in the encapsulation of
Although the interest for flavor ester encapsulation is still these compounds.
increasing and researchers working in this field are scarce. A few
studies evaluated the microencapsulation of benzyl benzoate in Acknowledgements
microemulsions of phospholipids (G. Sharma et al., 2015) and
microcapsules of calcium carbonate (CaCO3) (Wei, He, Yu, & Zou, The authors thank the financial support from CNPq (Conselho
2016) for usage as a topical drug in scabies treatment and anti- Nacional de Desenvolvimento Científico e Tecnolo gico), CAPES
mite activity, respectively. (Coordenaç~
ao de Aperfeiçoamento de Pessoal de Nível Superior)
Besides the microencapsulation, there are the utilization of and Universidade Federal de Santa Catarina (UFSC).
several types of systems as potential nanocarriers, such as solid
lipid nanoparticles, nanoemulsions and polymer nanoparticles. The
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