See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/339602667

ENZYMES IN THE PRODUCTION OF JUICES AND BEVERAGES

Article in WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES · March 2020

DOI: 10.20959/wjpps20203-15735

CITATIONS READS

0 1,133

1 author:

Michele Vitolo
University of São Paulo
170 PUBLICATIONS 2,797 CITATIONS

SEE PROFILE

Some of the authors of this publication are also working on these related projects:

Esterification of glycerol with short-chain fatty acids. View project

NAP - Núcleo de Apoio à Pesquisa em Fármacos e Medicamentos para Doenças Negligenciadas View project

All content following this page was uploaded by Michele Vitolo on 01 March 2020.

The user has requested enhancement of the downloaded file.

 WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES
Vitolo. World Journal of Pharmacy and Pharmaceutical Sciences
SJIF Impact Factor 7.632

Volume 9, Issue 3, 504-517 Review Article ISSN 2278 – 4357

ENZYMES IN THE PRODUCTION OF JUICES AND BEVERAGES

Michele Vitolo*

School of Pharmaceutical Sciences, University of São Paulo, Brazil.

Article Received on
ABSTRACT
10 Jan. 2020, Enzymes are biological catalysts with a large application in fruit juice
Revised on 31 Jan. 2020,
Accepted on 21 Feb. 2020 processing, winemaking, and brewing. Such processes cannot reach a
DOI: 10.20959/wjpps20203-15735 high-volume production, as demanded by the market, without using
enzymes (pectinases, cellulases, amylases, proteases etc.). Enzymes in

*Corresponding Author fruit juice processing have been used since the 1930s. They are pivotal
Prof. Dr. Michele Vitolo in the clarification of juices by eliminating mainly pectin and starch at
School of Pharmaceutical the fruit crushing step. In winemaking, enzymes play a role in reducing
Sciences, University of São
the splashing of grape juice during pressing and increasing the
Paulo, Brazil.
fermentation yield, whereas in brewing, they accelerate mashing,
improve fermentation yield, and avoid haze formation in beer during maturation (step before
packaging).

KEYWORDS: Enzymes, winemaking, brewing, fruit juice process.

INTRODUCTION
Enzymes are proteins that, in small amounts, can increase the rate of chemical reactions.
They are not consumed during the reaction, and their molecular structure remains unchanged
at the end of the reaction.[1] Specificity is a remarkable characteristic of enzymes, i.e., the
capacity of distinguishing and transforming one compound even when it is mixed with others
with a similar chemical structure. Enzymes can be used in soluble or insoluble forms.[2] In
fruit juice, winemaking and brewing processes (except for the beer maturation process, which
can be made with immobilized papain), enzymes are used in the soluble form.[3][4][5][6]

Enzymes are used in industry since the beginning of the twentieth century. In fruit juice
processing, the use of enzymes began in the 1930s, when pectinase and cellulase were
introduced in maceration of fruits and clarification of juices. Certainly, the development of
the juice industry - not only the huge volumes handled, but also the variety of fruit juices

www.wjpps.com Vol 9, Issue 3, 2020. 504

Vitolo. World Journal of Pharmacy and Pharmaceutical Sciences

found in the market (orange, apple, pear, strawberry, lemon etc.) – is due to the use of
enzymes. In winemaking and brewing, enzymes began to be used in the 1960s aiming to meet
the market demands for huge volumes of wine and beer. Winery requires enzymes to
accelerate the must fermentation and reduce the splashing, as well as increase the volume of
juice extracted during maceration. In brewing, the introduction of enzymes aims to improve
mashing (preparation of the wort) of malted barley (natural source of hydrolytic enzymes)
supplemented with cereals (soya, corn etc.). This mixture is needed to circumvent the lack of
malted barley to meet the huge volume of beer demanded by the market.

The aim of this work is to discuss briefly the use of hydrolytic enzymes in three important
large-scale food processes, i.e., brewing, winemaking, and fruit juice production.

Enzymes In Fruit Juices and Wine

The main problem of the fruit juice industry is to cope with the variability of fruits to be
processed. The optimization of the production implies processing the fruit at the lowest cost
during a short time while maintaining or improving the stability and the organoleptic quality
of the final product. Moreover, the broadening of the plant capacity is another aim.
Commonly, juices are commercialized as concentrates to decrease the volume, which
facilitates the storage and the transportation of the product, and to increase resistance against
microbial spoilage. These aims have been reached since the 1930s by using enzymes in the
juice processing.[7]

The fruit juice producer disposes, at any time and volume, of a wide range of specific and
partially purified enzymes, as well as blended enzyme preparations.

The main obstacle to extract the juice from a fruit is to destroy the cell wall, a formidable
resistant structure that surrounds plant cells. Its composition depends on the fruit variety,
agronomic and climatic conditions, and the method and duration of storage.

The plant cell wall is composed by middle lamella [outer layer of the cell wall composed of
soluble pectin and hemicellulose fractions (xylans, xyloglucan and arabinans); it is
responsible for the adhesion of adjacent cells binding to one another], primary wall [a layer
located below the middle lamella forming a network composed mainly of pectin (30%),
crystalline cellulose (40%), hemicellulose (20%), and glycoproteins (10%)], and the

www.wjpps.com Vol 9, Issue 3, 2020. 505

Vitolo. World Journal of Pharmacy and Pharmaceutical Sciences

secondary wall [promotes an extra reinforcement of cell wall; it is composed of cellulose,

hemicellulose, and lignin].[8][9]

The cell wall matrix is an intricate web resulting of the interaction of several polymers
(pectin, xylan, cellulose etc.).[7] In web, pectin is linked through proteins to xyloglucan and to
cellulose by hydrogen or covalent bonds. Xyloglucan is the main interlocking polymer,
though other non-cellulosic polysaccharides, such as galactoglucomannans and (1,3)--D-
glucans and glucuronoarabinoxylans, also appear as interlockers, but in lower amounts.
Moreover, other polysaccharides (arabinans, galactans, and branched arabinogalactans) of
various configurations and sizes are attached to the pectic backbone.[7]

Pectic substances are polysaccharides constituted by monomers of galacturonic acid and

rhamnose. The rhamnogalacturonan backbone consists of -D-galacturonan residues attached
by -1,4 glycosidic linkage, which are interrupted at 25-30 galacturonan units by the
insertion of -1,2 and -1,4-linked -L-rhamnosyl residues and alternating branches of
rhamnogalacturonan chains. Moreover, sugars such as L-arabinose, D-galactose, D-xylose
and L-fucose are attached to the side rhamnogalacturonan chains by glycosidic linkage to
carbon atoms 3 and 4 of the L-rhamnose units and carbon atoms 2 and 3 of the D-
[10][11]
galacturonic units. In despite of the great diversity in MW and the structure of pectic
substances, they can be divided into pectic acid (low degree of esterification) and pectinic
acid (high degree of esterification).

The main enzymes used in fruit juice industry are microbial pectinases, cellulases,
hemicellulases, and amylases.

ENZYMES
Pectinases
Pectinases are divided into esterases (PE) - hydrolyses the ester bonds of both pectic and
pectinic acids freeing alcohol (methanol, more commonly) - and depolymerases, which
disrupts the glycosidic bonds of the pectic polymer backbone (Figure 1). The depolymerases
that disrupt pectic acid and pectinic acid are called, respectively, pectic-acid-depolymerizing
(PAD) and pectin-depolymerizing (PD) pectinases. The cleavage of glycosidic bonds can be
by hydrolysis (proceeds using water) or trans-elimination (two hydrogen atoms belonging to
adjacent carbon atoms are eliminated without water) (Figure 2). Thereby, the pectinases
acting on pectinic acid/pectic acid by either hydrolysis or trans-elimination are called,

www.wjpps.com Vol 9, Issue 3, 2020. 506

Vitolo. World Journal of Pharmacy and Pharmaceutical Sciences

respectively, polymethylgalacturonases (PD-PMG)/polygalacturonases (PAD-PG) and

pectin lyases (PD-PL)/pectate lyases (PAD-PAL). If the glycosidic bond disruption takes
place as a random action pattern, then enzymes are called endopectinases (endo-PD-PMG,
endo-PD-PL, endo-PAD-PG and endo-PAD-PAL), but if the reaction goes on in a stepwise
release pattern beginning from one terminus of the polysaccharide chain, they are called
exopectinases (exo-PD-PMG, exo-PD-PL, exo-PAD-PG and exo-PAD-PAL).

Pectinases can be found in plants and microorganisms: PE (apple, pea, potato, Aspergillus
niger), endo-PAD-PG (avocado, papaya, Rhizopus sp., Kluyveromyces fragilis), exo-PAD-
PG (grapefruit, carrots, A. niger), endo-PAD-PAL (Bacillus polymixa), exo-PAD-PAL
(Erwinia sp., Clostridium sp.), endo-PD-PL (A. niger, Erwinia sp.), and endo-PD-PMG (A.
niger).[11]

Methods for assaying pectinases activities vary according to the type of enzyme.
Pectinesterase activity – one unit of PE is defined as the amount of enzyme that liberates one
micromole of free carboxyl groups or methanol per minute under specified assay conditions –
is determined by titrating the free carboxyl groups and/or the free methanol by gas liquid
chromatography.[11][2] Pectin lyase (PD-PL) and pectate lyase (PAD-PAL) activities are
measured by monitoring the increase in light absorbance ( = 232 nm) caused by the
formation of the double bound at the non-reducing ends of the molecules. One unit of lyase
liberates one micromole of unsaturated products per minute under specified assay
conditions.[11][13][14] Polygalacturonase activities (PG and PMG) are determined by
measuring the increase in reducing groups by using the Somogyi-Nelson’s reagent. One unit
of PG/PMG is defined as the amount of enzyme that liberates one micromole of reducing
groups per minute under specified assay conditions.[11][15]

www.wjpps.com Vol 9, Issue 3, 2020. 507

Vitolo. World Journal of Pharmacy and Pharmaceutical Sciences

PECTINASES ESTERASES (PE)

DEPOLYMERIZING

PD PAD

EXO-
PD-PL

PD-PMG PD-PL PAD-PG PAD-PAL

ENDO- EXO-
PD-PL PAD-PG

ENDO EXO- ENDO- EXO- ENDO-

-PD-PMG PD-PMG PAD-PG PAD-PAL PAD-PAL

Figure 1: Classification of pectinases. PD = pectin-depolymerizing pectinase; PAD =

pectic-acid-depolymerizing pectinase; PD-PMG = polymethylgalacturonases; PD-PL =
pectin lyases; PAD-PG = polygalacturonases; PAD-PAL = pectate lyases.

Figure 2: Splitting of glycosidic bonds into pectic substances by hydrolysis (A) and by
trans-elimination (B).

OTHER ENZYMES
Rhamnogalacturonase– The optimal pH is between 3.0 and 4.0. – It hydrolyses the pectic
substances between rhamnose and galacturonic residues (pectic elbow) in branched regions
of the pectic substance after its deacetylation. The main source is Aspergillus aculeatus.

The arabinose polymers (arabinnans, arabinoxylans, arabinogalactans) are hydrolyzed by

arabinofuranosidases (endo-arabinase and exo-arabinase). The degradation of these
polymers avoids turbidity in the juice produced (mainly apple and pear juices).

www.wjpps.com Vol 9, Issue 3, 2020. 508

Vitolo. World Journal of Pharmacy and Pharmaceutical Sciences

Hemicellulases represent a group of enzymes which hydrolyses the hemicellulose in the cell
wall. Galactanases, xylanases, xyloglucanases and mannanases are the most important
enzymes of this group.

The so called cellulases constitute a group of enzymes which act synergistically. The
cellulolitic complex is formed by endo-glucanase (cleaves randomly -1,4 glycosidic
bonds), exo-glucanase (hydrolyses -1,4 glycosidic bonds from the reducing extremity of the
polysaccharide chain), cellobiohydrolase (acts after endo-glucanase, freeing cellobiose by
cleaving the glycosidic bonds from the non-reducing terminus of the oligosaccharide chain),
and cellobiase (also called -glucosidase, it splits cellobiose into two molecules of glucose).

Amylases are used to avoid the cloudiness of the final product due to the precipitation of the
starch naturally present in the unripe fruit. The most used are glucoamylase (pHoptimal = 4.3;
temperature between 55oC and 60oC) and microbial α-amylase (pHoptimal between 3.0 and
4.0; temperature between 70oC and 75oC).

Figure 3 shows a typical fruit juice processing plant. The addition of enzymes in the holding
tank aims to eliminate the insoluble pectin, meanwhile in the clarification tank, the addition
of enzyme aims to reduce juice viscosity to remove the residual starch (avoiding juice
cloudiness) and to facilitate the flocculation of insoluble substances.

CRUSHING MILL HEAT EXCHANGER

SEPARATOR PRESS HOLDING TANK

CLARIFICATION TANK SEPARATOR

CONCENTRATOR FILTER PRESS

STORAGE TANK PACKAGING SECTOR

Figure 3: Steps of a juice processing plant. Enzymes are added in the holding tank (3-20
g of pectinase and 0.2-2 g of cellulase per 100 Kg of fruit) and in the clarification tank
(1.5-3.0 g of pectinase and 0.5-2.0 g of amylase per 100 L of juice).[7]

www.wjpps.com Vol 9, Issue 3, 2020. 509

Vitolo. World Journal of Pharmacy and Pharmaceutical Sciences

The clarification process is biphasic. The pectinase hydrolyses pectic substances (1st phase)
followed by the electrostatic agglutination of disarranged micelles in suspension in the juice
(2nd phase).

The expectation of fruit juice industry regarding enzymes relies on increasing the availability
of pectinases resistant to the acid medium, so that they can be used in citric fruit processing.
The availability of naringinase, a debittering enzyme for citric juice fruit production, is
another good perspective for enzymes in this industry.

WINE
Though wine is a fermented alcoholic beverage, its production, before fermentation, involves
operations similar as those of fruit juice processing. Winemaking is a biotechnological
process operated by humans at least from 3,000 years BC. Figure 4 shows the main steps of
the production of wine.

The main objective for the winemaker is to obtain a product with a high quality and fitness by
keeping and enhancing the natural aromas of the grape berry, and to generate new aromas
through controlled fermentation of the grape juice and aging the wine at controlled
temperature, humidity, and storage in proper barrels.

Although winemaking still involves some degree of empiricism, the high market demand
requires a huge production of stable wines of high quality produced in a short period. The
long aging periods are reserved only for special non-popular wines, which, of course, are
expensive goods.

The winemaker, as the juice producer, must deal with the cell walls that surround grape cells
rich in juice (the substrate to be fermented). For instance, one kilogram of Chardonnay grape
cell wall contains about 4 g of rhamnose, arabinose, galactose, mannose, uronic acid,
methanol, acetic acid, proteins, and xylose.[16] The grape pectin is a highly branched polymer,
presenting a methylation level of about 70%.

During grape maturation, the pectin is hydrolyzed by endogenous pectinases, whose activities
are too low to have a technological effect. Thereby, the winemaker, to meet the high demand
for unsophisticated wines, is obliged to use exogenous pectinases in order to obtain a better
initial extraction of the various must components (juice, color, flavor etc.), thus improving
yield. When added to grape must, enzymes hydrolyze the soluble high molecular weight

www.wjpps.com Vol 9, Issue 3, 2020. 510

Vitolo. World Journal of Pharmacy and Pharmaceutical Sciences

substances, such as pectin and arabinogalactans, improving clarification and filtration. Pectin
– whose amount in grapes depends on the grape variety, maturity and the technology used
after harvest -, in presence of water and sugars at an acid pH, forms a viscous gel that settles
particles in suspension and makes clarification almost impossible.

Pectin is the main substance to be removed from the must before the fermentation process
(Figure 4). This is accomplished by using pectinases before pressing crushed grapes, so that
the pressing yield is improved as well as the clarification and the filtration of the latter.
Commercial pectinases are from Aspergillus niger with secondary activities such as
glycosidases (mainly, α-L- rhamnosidase, α-L-arabinosidase, -D-apiosidase and -D-
glucosidase; all of them related to the release of the aroma from the grape) and proteases
(important for preventing the haze formation; they can provide amino acids that serve as
nutrients for yeasts and malo-lactic bacteria, which can lead to the improvement of the
organoleptic quality of the wine).

GRAPES MACERATION AND

PRESSING FERMENTATION

CRUSHING
PRESSING
MUST CLARIFICATION

WINE CLARIFICATION
FERMENTATION

AGING
WINE

FILTRATION AGING FILTRATION

WHITE WINE FILTRATION RED WINE

Figure 4: Diagram of the steps for obtaining red and white wine in presence of pectinase
(●), -glucanase (●) and/or proteinase (●).

Often grapes are contaminated with Botrytis cinerea (common fungus in grapevines), which
produces a glucan-like gum with -1,6-linked side chains attached to a -1,3 linked
backbone. This gum, once dissolved in the grape juice, causes problems in wine clarification
and filtration, which can be circumvented by adding glucanases.

www.wjpps.com Vol 9, Issue 3, 2020. 511

Vitolo. World Journal of Pharmacy and Pharmaceutical Sciences

Enzymes already available in the market are enough for the enologist to solve the main
problems affecting winemaking. However, genetically modified, DNA-edited or synthetic (in
the future) Saccharomyces cerevisiae, for having the capability to produce pectinases and
release specific components such as color or aromas, would allow coupling clarification with
fermentation, leading to a shorter winemaking process.[17]

BREWING
Brewing is a traditional process to produce beer that has been developed along the centuries.
Beer is one of the most ancient biotechnology products, whose initial processes were set
around 3,500 B.C.

The main raw materials used to brew beer are water, malted barley (supplemented or not with
non-malted cereal adjuncts), hops, and yeast.

The brewing process involves extracting and breaking down the carbohydrates of the malted
barley and/or non-malted cereals to make a sugar solution (also containing essential
nutrients), and using this as a source of nutrients for anaerobic yeast growth to break down
the simple sugars, releasing energy and producing ethanol as a metabolic byproduct.

The major biological changes in the brewing process result from the action of enzymes
present in barley and those produced by the yeast during fermentation. The rest of the
brewing process involves heat exchange, separation and clarification, which do not promote
significant changes in the beer organoleptic characteristic. Barley can produce all the
enzymes needed to degrade starch, -glucan, pentosans, lipids and proteins, which are the
major compounds of concern to the brewer. The main enzymes in the malt are -amylase, -
amylase, -glucanase and carboxypeptidase, which are stable up to 68oC, 64oC, 62oC and
58oC, respectively.[18]

The main steps involved in the brewing process are (Figure 5):[19]

A) Malting: malt barley - Low percentages of nitrogen and -glucan - is the preferred cereal
for producing the must, which will be fermented into beer. Malting involves Steeping (the
cereal is saturated with water up to a moisture of 45% (w/w)), Germination (a 3-5-day
process, during which all natural enzymes are synthesized and the main endosperm
constituents - starch, proteins, -glucan and pentosan gums - are decomposed; their

www.wjpps.com Vol 9, Issue 3, 2020. 512

Vitolo. World Journal of Pharmacy and Pharmaceutical Sciences

derivatives will become the nutrients of the fermentable must. The germination is carried out
at 15oC, relative humidity of 100%, and air is blown continuously along the process), Kilning
(a soft drying of germinated malt aiming to reduce the moisture to 3-6% (w/w) of the whole
malt. It takes usually between 24 h and 48 h, during which air smoothly heated is blown
through the germinated malt), and malt dressing (after kilning, the malt is left maturing for
at least 21 days, in which the residual moisture distributes uniformly throughout the material.
During malting, the barley natural enzymes are activated, leading to the softening of
endosperm cell wall – due to the hydrolysis of -glucans and the protein matrix surrounding
the starch granules – and the proper germination of the corn);
B) Mashing: it is the programmed cooking of the ground, germinated malt from which
results the wort (malt-derived sugar solution) rich in reducing sugars and nitrogenous
compounds. The cooking program consists on a gradual increase in the mash temperature to
promote the sequential action of malt enzymes (carboxypeptidase  -glucanase -
amylase  -amylase) on proteins and carbohydrates present in the cereal. The pattern of
temperature variation depends on the quality and composition of the raw material. The
temperature varies between 58oC (kept for 20 min to enhance the carboxypeptidase activity)
and 65oC (kept for 40 min to enhance α-amylase and -amylase activities; at this temperature,
the starch granules are swelled and gelatinized). As a rule, the higher the temperature
escalation, the lower the quality of the starting modified cereal;
C) Fermentation: at this phase, the wort is introduced into a fermenter and inoculated with
an adequate yeast strain so that after several hours of discontinuous fermentation, the sugar
present in the medium is converted under anaerobic conditions into ethanol. The yeast cells
are removed from the fermented broth, which is left to stabilize for a period depending on the
type of beer desired. Sometimes, there is interest to produce a beer with a special
characteristic, for example, a low calorie beer (or "light beer"). In a regular beer, about one-
third of the wort is constituted by soluble dextrins (starch -(16) branch linkages) derived
from incomplete starch hydrolysis. Dextrins are not fermentable carbohydrates therefore they
accumulate in the finished beer, increasing its caloric content. By introducing glucoamylase
into the wort, the dextrins are hydrolyzed, freeing more glucose to be fermented by the yeast.
At the end, the obtained beer has a higher alcoholic degree but a lower residual carbohydrate
concentration and consequently lesser calories. If the high-alcohol beer is diluted to meet the
legal alcohol concentration in regular beer, the final carbohydrate concentration decreases
even more. At this phase, if the wort presents low fermentable sugars concentration and/or

www.wjpps.com Vol 9, Issue 3, 2020. 513

Vitolo. World Journal of Pharmacy and Pharmaceutical Sciences

high viscosity, the brewer can add -amylase and/or -glucanase;

D) Beer Stabilization: at this phase, the haze-forming compounds (hydrophobic proteins,
flavonoids, polyphenols, ions, and carbohydrates) are removed. Oxidation plays an important
role in the polymerization of polyphenols to produce an irreversible haze. The amount of
haze varies depending on factors such as the type of beer and increases with age, exposure to
oxygen, and agitation during shipping. Eventually, the beer becomes hazy even at warm
temperatures and sediment may appear. The haze is not desirable for aesthetic reasons: it
resembles cloudiness due to microbial spoilage. To circumvent this problem, papain (a
mixture of cystein proteases from Carica papaya) is added (1-5 g/100 L of beer) to the
fermented broth either during transfer between the fermenter and the maturation tank or into
the maturation tank itself. Before packaging, the beer is pasteurized to increase the shelf-life
and inactivate papain.

MALT MILLING CASKS

CEREAL ADJUNCTS

CANS
WATER
KEGS

PACKAGING BOTTLES
MASHING

BYPRODUCTS
FILTRATION

Malt dust
LAUTERING

HOPS PASTEURIZATION Spent grains

BOILING Protein debris

MATURATION Hops debris

CLARIFICATION
YEAST CO2

Spent filter aid

COOLING FERMENTATION

Figure 5: Schematic of brewing process: milling (particle size reduction), mashing

(hydrolysis of starch and sugars), lautering (separation wort from solids), boiling (boiling
with hops), clarification (whirlpool), fermentation (sugar conversion in ethanol),
maturation (storage and stabilization). ENZYMES: papain (■), bacterial neutral protease
(■), fungal pentosanase (), fungal glucanase (), bacterial glucanase (), bacterial -
amylase (●), fungal -amylase (●), bacterial debranching enzyme (), and glucoamylase
(●).

www.wjpps.com Vol 9, Issue 3, 2020. 514

Vitolo. World Journal of Pharmacy and Pharmaceutical Sciences

Due to the huge amount of beer commercialized worldwide, there is often shortage of good
barley malt in the market. In this case, the brewer must use a blend of barley malt with other
cereals (soy, rice, cassava, bran etc.), which leads to a low-quality wort, characterized by a
low concentration of reducing sugars and nitrogenous compounds. Therefore, the brewer
adds to the mash exogenous enzymes, mainly carbohydrases (glucanase, α-amylase, -
amylase and cellulase) and proteases.

The use of non-malted adjuncts – which are poor in endogenous enzymes – in brewing
formulations is required when some particular flavor, aroma and/or processing advantages are
envisaged by the brewer. Then, the addition of exogenous enzymes is obligatory.

The future trends regarding the use of enzymes in brewing relies on the development of
immobilized papain (haze removing would occur during the transfer of the fermented broth
from the fermenter to the maturation tank, which, in turn, should become an reservoir tank
from which the beer would be bottled), engineered yeast (high fermentation capability
associated with glucoamylase synthesis for the production of “light beer”), and thermostable
proteases and glucanases.

CONCLUSION
Fruit juice, wine and beer processing require mainly hydrolytic enzymes (pectinases,
proteases and carbohydrases) in order to remove undesirable polymers such as pectin,
cellulose, starch, hemicellulose, among others. The future trend for enzymes for beer
production relies on the availability of immobilized papain and thermostable proteases and
glucanases, whereas in fruit juice production, acid-resistant pectinase and naringinase are
required (specifically for citric juice production). Regarding winemaking, the enzymes
available are enough to meet all needs of the process. Moreover, the aim of winery and
brewing is to improve the fermentation using engineered Saccharomyces cerevisiae. In
winemaking, the use of fermentable yeast capable of producing pectinase could allow
coupling clarification with fermentation, leading to a shorter time of the process. In brewing,
yeast with high fermentation capability, associated with glucoamylase synthesis to produce
“light beer,” may be a great commercial interest.

Funding: This study was supported by the National Council for Scientific and Technological
Development – CNPq (grant no. 303082/2015-1).

www.wjpps.com Vol 9, Issue 3, 2020. 515

Vitolo. World Journal of Pharmacy and Pharmaceutical Sciences

REFERENCES
1. Vitolo M. Brief review on enzyme activity. World Journal of Pharmaceutical Research,
2020; 9(2): 60-76.
2. Vitolo M. Calcium-alginate beads as carriers for biocatalyst encapsulation. World Journal
of Pharmaceutical Research, 2019; 8(10): 1-17.
3. Gomaa AM. Application of enzymes in brewing. Journal Nutrition of Food Science
Forecast, 2018; 1(1): 1-5.
4. Mojsov K. Use of enzymes in winemaking: a review. International Journal of Marketing
and Technology, 2013; 3(9): 112-127.
5. Ramadan MF. Enzymes in fruit juice processing. In: Enzymes in Food Biotechnology.
Kuddus M (Ed). London, Academic Press, 2019; 45-59.
6. Vitolo M. Enzymes and applications. In: Pharmaceutical Biotechnology. São Paulo,
Edgard Blücher, 2015; 247-287.
7. Grassin C, Fauquembergue P. Fruit juices. In: Industrial Enzymology. Godfrey T, West S
(Eds). New York, MacMillan Press Ltd, 1996; 227-264.
8. Selvendran RR. The chemistry of plant cell walls. In: Dietary Fibre. London, Applied
Science Publishers, 1983; 95-147.
9. Strasser GR, Amadó R. Pectic substances from red beet. Carbohydrate Polymers, 2001;
44: 63-70.
10. Walsh G, Headon D. Protein Biotechnology. London, Willey Publishers, 1994.
11. Pilnik W, Voragen ACJ. Pectic enzymes in fruit and vegetable juice manufacture. In:
Enzymes in Food Processing. Nagodawithana T, Reed G (Eds). San Diego, Academic
Press Inc., 1996; 363-399.
12. McFeeters RF, Armstrong SA. Measurement of pectin methylation in plant cell walls.
Analytical Biochemistry, 1984; 139: 212-217.
13. Edstrom RD, Phaff HJ. Purification and certain properties of pectin-transeliminase.
Journal Biological Chemistry, 1964; 239: 2403-2407.
14. McMillan JD, Phaff HJ. Exopolygalacturonate lyase from Clostridium multifermentans.
Methods in Enzymology, 1966; 8: 632-635.
15. Spiro RG. Analysis of sugars found in glycoproteins. Methods in Enzymology, 1966; 8:
3-26.
16. Grassin C, Fauquembergue P. Wine. In: Industrial Enzymology. Godfrey T, West S
(Eds). New York, MacMillan Press Ltd, 1996; 375-383.

www.wjpps.com Vol 9, Issue 3, 2020. 516

 Vitolo. World Journal of Pharmacy and Pharmaceutical Sciences

17. Ostrov N. Design, synthesis, and testing toward a 57-codon genome. Science, 2017; 18:
749-760.
18. Power J. Enzymes in brewing. In: Enzymes in Food Processing. Nagodawithana T, Reed
G (Eds). San Diego, Academic Press Inc., 1996; 439-457.
19. Rourke TO. Brewing. In: Industrial Enzymology. Godfrey T, West S (Eds). New York,
MacMillan Press Ltd, 1996; 105-131.

www.wjpps.com Vol 9, Issue 3, 2020. 517

View publication stats