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To cite this article: Horacio Guzmán‐Maldonado, Octavio Paredes‐López & Dr. Costas G. Biliaderis (1995): Amylolytic enzymes
and products derived from starch: A review, Critical Reviews in Food Science and Nutrition, 35:5, 373-403
This article may be used for research, teaching, and private study purposes. Any substantial or systematic
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Critical Reviews in Food Science and Nutrition, 35(5):373^M)3 (1995)
Referee: Dr. Costas G. Biliaderis, Dept. of Food Science and Technology, School of Agriculture, Aristotle University, Thessaloniki,
Greece
ABSTRACT: This review provides current information on starch and its molecular composition, common and
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potential sources, and manufacturing processes. It also deals with the five groups of enzymes involved in the
hydrolysis of starch: the endo- and exoamylases, which act primarily on the α-1,4 linkages; the debranching
enzymes, which act on the α-1,6 linkages; the isomerases, which convert glucose to fructose; and the cyclodextrin
glycosyltransferases, which degrade starch by catalyzing cyclization and disproportionation reactions.
This work mainly discusses the enzymatic processes for the manufacture of maltodextrins and corn syrup
solids, including the production, both batch and continuous, of glucose syrup, and the processes to obtain
sweeteners, such as maltose and 42, 55, and 90% high-fructose corn syrups. It highlights the novel production of
Schardinger's dextrins: the α-, β-, and γ-cyclodextrins, consisting of six, seven, and eight glucose monomers,
respectively.
New products are emerging on the market that can serve as fat and oil substitutes, moisture-retention
compounds, crystal-formation controllers, stabilizers for volatile materials like flavors and spices, or products for
the pharmaceutical industry. As a result, particular attention is given to functional properties and applications of
the above-cited compounds.
1040-8398/95/$.50
© 1995 by CRC Press, Inc.
373
commonly used sweetener has been sucrose, but hydroxymethyl position of certain glucose moi-
in the 1970s, a highly significant change took eties via OC-D-1,6 linkages.19-64 Although it is now
place as a starch-derived sweetener, fructose syrup, accepted that amylose is not completely linear,74
came into common use. Methods for chemical its solution properties are typical of those of a
separation of glucose and fructose have been de- linear polymer.19
veloped and enrichment of the enzymatically pro- More than half of all starch production goes
duced syrup to 90% fructose is practiced by sev- to the manufacture of food and feed products,
eral producers today. either as such or in pregelatinized and chemically
This article reviews the amylolytic enzymes, modified form.66-119 However, modified starches
their types and action patterns, and the production are used in the paper, textile, soap, laundry, cos-
of novel derivatives of starch that are influencing metic, and pharmaceutical industries; in fireproof-
both the public and the food industry. In particu- ing preparations; as explosive- and fuel-binding
lar, emphasis is placed on the functional proper- agents; and as flocculating agents for water treat-
ties and food applications of these products. ments and in the building industry.21-66-91 These
industries, especially the paper and board indus-
try, consume a remarkable amount of starch,
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A. Introduction
B. Common Sources
The cold water-insoluble starch granule forms
the major constituent of all cereal grains. It is a 1. Maize
reserve substance composed entirely of oc-D-glu-
cose for most higher plants and constitutes an Of the common starches, regular corn, waxy
energy source essential for many organisms, es- corn, and high amylose corn are by far the most
pecially humans.64 From a nutrition standpoint, important sources. Nevertheless, only about 5%
starch is the major component of the diet in all of the annual world maize crop is used for the
human populations. However, in our culture, the manufacture of maize starch.20 About 70% of the
main purposes of starch utilization remain aes- maize starch produced is converted into corn syrup,
thetic rather than nutritional. This biopolymer high fructose corn syrup, and dextrose.20
constitutes an excellent raw material for modify- Corn starch granules are medium sized and
ing food structure and consistency.19 round or polygonal in shape. Starch content of
The commercial and technological uses of corn ranges from 68 to 74%. The amylose content
starch are extremely numerous; this arises from ranges from about 80% in high amylose to about
its unique character as it can be used directly as 1% in waxy starch.20-159
intact granules, in the dispersed form, as a film Maize starch has a wide variety of industrial
dried from a dispersion, or as an extrudate powder applications, with uses ranging from thickening
after conversion to a mixture of oligosaccharides and gelling agents in puddings and fillings, to
or via hydrolysis and isomerization.66-91-177 As a molding for confections. Chemical modification
result, the academic aspects of the subject have of maize starches can be used to extend the func-
received much attention. The most important ad- tional characteristics to satisfy a wide range of
vance in the study of starch was the realization industrial requirements.21-159-168
that the starch granule was not chemically homo-
geneous,41-66-189 thus separation could be made into
the simpler component amylose, an essentially 2. Potato
linear polymer of glucopyranose units linked
through OC-D-1,4 linkages, and amylopectin, a About 3% of the world crop of potatoes is
branched polymer containing chains with a short used for the production of potato starch. Potato
degree of polymerization (DP = 20 to 25 starch granules are oval in shape, with pronounced
glucopyranose residues) linked to the C-6 oyster shell-like striations around an acentrically
374
placed hilum.20 Potato starch has the largest gran- from fiber, grits, pulp, or gluten. Basically, it is a
ules of any commercial starch, and its products process that separates germ, oil, protein, and starch
are used in the food, paper, textile, and adhesive using mainly screening, centrifugation, and plenty
industries.96-183 of water without any concomitant degradation of
the starch. The starch suspension free of protein
and fine fiber material is dewatered, then dried,
3. Wheat and finally sifted.5-21-51
and the larger, lenticular granules.20 starch content, with 27% amylose.73'190 Triticale
Wheat starch is used in the baking industry has 54% starch, with an amylose content of ap-
and in the production of adhesives as well as proximately 23%.18'73 Wild rice has 60 to 65%
having applications in the confection and canning starch, with 16.8 to 23.9% amylose.82 Amaranth
industries.20 has 48 to 69% starch, with an amylose content of
0 to 21.8%.134138'140'157'171'172 Some efforts have
been made to study the potential use of amy-
4. Rice lolytic hydrolysis for these materials, especially
with amaranth14'61-69-70-139 and cassava16-43-44'61
Most of the material for rice starch manufac- starches.
turing consists of rice grains that are damaged or
broken in mills that produce rice for food.20 Rice
starch has the smallest granules of all commercial III. AMYLOLYTIC ENZYMES
starches; the granules tend to aggregate within the
amyloplast in spherical clusters and each amylo- A. Introduction
plast contains 20 to 60 small polyhedral gran-
ules.83 Rice starch is used as laundry starch, in The term enzyme, which means "in yeast,"
cosmetic dusting powders, and as a thickener in gives only a few clues as to what the sources of
puddings and ice cream.81 enzymes are.40 Primitive man relied on plants and
certain organs of animals for their enzymatic roles
in brewing, cheesemaking, and other food pro-
C. Manufacturing Processes cesses. Malting was done by the ancient Chinese
1000 years before Christ; it was used to manufac-
Some variations in procedure may be required ture "Hung Fan," meaning "sweet from cereal."30
in isolating starches from different sources (Fig- It is interesting to note that the first enzymes to be
ure 1). It is necessary that starch be softened by recognized as specific biocatalysts were those
some type of process: cleaning by steeping with capable of hydrolyzing starch.51
S02 in maize,5-21'51 pulping in potato tubers,20 and The development of the starch/wet-milling
dough mixing in wheat.20 Also, it is necessary to industry can be traced back to the French general
carry out the extraction process in the presence of Napoleon in the eighteenth century. Because of
mercuric chloride (0.01 M) in order to inhibit Napoleon's continental blockade, Europe found
endogenous starch-degrading enzymes.20'21 itself without its traditional sources of sugar.
Different steps of Figure 1 are repeated until Therefore, rewards were offered for methods that
the material that passes through the sieves is free could result in a suitable substitute.51 While work-
375
pre-«illin$ fine entrifuoinj
cleaning STARCH
•—• separation — —» nillUg — cyclone — —t.
steeping
uasMrig separation
Fulp Effluent
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"ft" STARCH
— —i
screening
dough gluten gluten centrifuging TSIATCH
FIOOR nixing dewelopnentl washing cuclone
separation
Solubles
ing on this problem, the German chemist Kirchhoff A. oryzae produced a mixture of amylases. The
discovered that starch boiled with acid and then identification and use of these enzymes enabled
neutralized could be converted into sugar.109-161 production of syrups not possible with acid.51
However, the nonspecific catalytic action of the The real breakthrough in industrial terms came
acid resulted in the formation of undesirable re- in the mid-1960s, with the development of an
version products and color. Furthermore, the syrup amyloglucosidase, which made it possible for
had a relatively high salt content.142-166 Those starch to be completely hydrolyzed to glucose.108
undesirable characteristics of the syrups were A full enzymatic starch hydrolyzate was not
caused by acid hydrolysis and enzymes are now achieved until the high-temperature stable bacte-
supplementing or replacing that process.51 rial amylase from Bacillus licheniformis was in-
In 1833, a Frenchman, Payen, succeeded in troduced in 1973. The use of this enzyme enabled
isolating the active component in malt, which he production of syrups containing up to 98% glu-
called diastase. Takamine, in 1894, developed the cose, which is an ideal substrate for glucose
idea of using an enzyme mixture from Aspergil- isomerase to convert glucose to fructose.24-54-104-105
lus oryzae for consumption as a digestive aid for The worldwide market for industrial enzymes
Asian populations who had difficulties in digest- in 1990 has an estimated value of $625 million,
ing rice (starch).51-127 It was discovered that with the U.S. as the largest sector. About 62% of
376
the enzymes produced find their way into food which are known as "filtration enzymes" that
applications. In the food industry, the largest use positively affect the filterability of the syrups.93-95
of enzymes is in starch processing followed by The other group, named proteinases, is contained
cheese production, fruit and vegetable juice pro- in industrial preparations of starch-degrading en-
cessing, baking, and brewing.9-40 zymes. These release starch from protein-starch
The screening of microorganisms and plants complexes.55-60
has become standard procedure for yielding im-
proved forms of known enzymes. Recent advances
and creative approaches are demonstrating that 2. Endo-enzymes
enzymes can catalyze reactions under conditions
previously thought to be too extreme. Rapid These enzymes cleave the a-1,4 glycosidic
progress is being made toward fine-tuning many bonds in amylose, amylopectin, and related
of the diverse techniques that are required for polysaccharides but not the a-1,6 bonds in amy-
successful gene cloning and protein engineering lopectin.130 The products of hydrolysis are oli-
of amylolytic enzymes.25-76'102-141-178'179-180 gosaccharides of varying chain lengths and have
the cc-configuration on the Cl of the reducing
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377
« TABLE 1
3 Classification of Enzymes Involved in the Commercial Production of Starch Syrups, Maltodextrins and
Cyclodextrins
^•» *v w • • • VB p • •
Common Substrate
Type name Source specificity PH Temp. (°C)
B tD D 0 1
J 0
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G 0 G A \
1
0 • \
o-o-o-o-o-o^ 0 \
B D
0
\ •
starch molecules begin to reassociate into an or- M27, shows two peaks for pH optima at 6.5-7.0
dered structure)12'118 is greatly reduced. Another and 8.5-9.0, a temperature optimum at 85 to 90°C,
fact involves the requirements for Ca2+ ions for and an activation energy 20.4 times lower than
full stabilization of the enzymes. B. licheniformis that reported for the enzyme from another strain
needs only 5 ppm of Ca2+, whereas B. subtilis of B. licheniformis.136-141 A higher reaction rate
needs 150 ppm for saturation level.166 A reduction due to lower activation energy is thus offered by
in the Ca2+ concentration required for enzyme this enzyme.
stabilization and its removal later in refining of- The action pattern of B. licheniformis en-
fered economic advantages.165 zymes generates G5 (chain length expressed as
Three other thermostable a-amylases have number of glucose units) and larger products ini-
been at least partially purified. One from tially, followed by later production of G3, G2, G^
B. licheniformis MV10 is inhibited by calcium and G4 in descending order. The hydrolysis rate
ions and has an optimum temperature >80°C.97 decreases sharply with decreasing chain length;
Another from a mutant of B. licheniformis is ca- G2 and G3 are not attacked at all.148'165 The Michae-
pable of low pH (5.5) liquefaction.7 If a lower lis-Menten model was found to be sufficient for
operating pH can be achieved, then chemical ad- describing the kinetics at low starch concentra-
dition and costs for ion exchange are significantly tions; however, a modified first-order model was
reduced. The third enzyme, from B. licheniformis required at high concentrations.92 There is no
379
evidence of substrate inhibition at high starch in starch hydrolysis: the glucogenic and maltogenic
concentration.166 exoamylases.51
The other group of endoamylases, thermo- The glucogenic exoamylase, amyloglucosidase
labile a-amylases, is derived from fungal sources, or glucoamylase (EC 3.2.1.3) is fungal in origin
usually A. oryzae (Table 1). In the starch industry, (Table 1). This enzyme attacks a-1,4 linkages from
their use is mostly confined to the saccharifica- the nonreducing end, releasing D-glucose molecules
tion reaction. Products of this reaction are mainly in the P-configuration.109 Amyloglucosidase re-
maltose and maltotriose, and thermolabile a-amy- moves D-glucose molecules up to the point of
lases are hence referred to as maltogenic in action branching in the amylopectin molecule; hydrolysis
(Figure 2).62'108'148 Units of activity are defined as may also proceed on the a-1,6 bond, but much
for thermostable a-amylases, but under different more slowly than on a-1,4 bonds.108 Activity is
conditions, especially pH because, unlike bacte- usually stated in units per liter and is based on the
rial enzymes, fungal enzymes are usually more production of glucose (or reducing sugars) from a
stable and active under acidic conditions.51 defined substrate.51 This enzyme is one of the most
Properties of immobilized a-amylases can be important industrial enzymes and has worldwide
strongly influenced by diffusion limitations, which applications.2-108'109 The most important use of
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are aggravated by the high molecular weight of glucoamylase is in the production of high-glucose
the substrate and high solution viscosity. If an syrups (96 to 98% glucose)2-16 and high-fructose
immobilized a-amylase were packed in a col- syrups (55% fructose).2'104
umn, usually the most efficient procedure, high At high glucose concentrations, amylo-
pressure drops at any reasonable flow rate would glucosidase repolymerizes glucose molecules in a
also be expected.148 reaction called reversion, forming maltose and
Genetic-engineering techniques have opened isomaltose.151 This phenomenon is more pro-
new avenues for production of valuable enzymes. nounced at high substrate and high enzyme con-
One microorganism of interest, B. subtilis centrations.104 The polymerization reaction of glu-
TN106(pAT5), is a fast-growing bacterium, indi- cose can also be catalyzed by another enzyme,
cating very efficient glucose utilization. Further- transglucosidase (EC 2.4.1.24), which is often
more, batch culture experiments revealed that present in crude amyloglucosidase preparations.109
a-amylase synthesis and secretion were initiated Amyloglucosidase acts synergistically with
in the early part of the growth phase.180 a-amylase in raw starch hydrolysis.4849 The syn-
The a-amylase gene (amy) from Streptomy- ergistic action is retarded not only by a decrease
ces griseus IMRU 3570 was subcloned into in the molecular weight of the substrate but also
Brevibacterium lactofermentum. The amy gene by an increase in the concentration of the sub-
was expressed efficiently when the promoter of strate.48
the kanamycin resistance gene was inserted up- Several studies have been done on the immo-
stream of the promoterless amylase gene.25 Liu bilization of amyloglucosidase on a variety of
et al.102 reported an overproduction of a-amylase supports by a variety of methods.29-47-144'145 As a
in recombinant Escherichia coli. rule, immobilization brings about some changes
in enzyme properties such as kinetic, thermal sta-
bility, and optimum working conditions. More-
3. Exoamylases over, for immobilized enzymes, the restraint in
the diffusion of substrate molecules to its site of
These enzymes act on the same substrates as reaction becomes an important factor. When
endoamylases but in a different way. Some of amyloglucosidase was immobilized on DEAE-
them are able to cleave a-1,6 linkages (Figure 2), cellulose, there was an increase in K,,, and a de-
but the reaction rate is slow compared with their crease of Vmax for the hydrolysis of starch, which
hydrolysis of a-1,4 bonds. Exoamylases act ex- could be partially explained by diffusion limita-
ternally on substrate bonds from the nonreducing tions.144 On the other hand, thermostability of
end and produce low molecular weight products. amyloglucosidase increases with an increase in
Currently two types of exoenzymes are important the amount of enzyme adsorbed.145
380
Amyloglucosidase immobilized on celite is Saha and Zeikus156 have discovered a highly
as efficient as the soluble form in catalyzing hy- thermoactive (75 to 85°C) Clostridium thermo-
drolysis of maltose. However, it is less efficient sulfurogenes p-amylase. This enzyme makes it
than the soluble enzyme in hydrolyzing 30% (w/w) possible to saccharify liquefied starch at higher
maltodextrin, giving a maximum dextrose equiva- temperatures than have been used in the past.
lent (DE) value of 96.0% instead of 98.2%.29 Immobilization of P-amylase on alginate,
Such a low DE may result from the fact that the agarose, or ion-exchange carriers has been re-
accessibility of large oligosaccharide molecules ported.35 It was observed that the type of carrier
to the active site of immobilized enzyme is re- and the source of the P-amylase are important
tarded by the limited space inside the pore, and factors for the required capacity of the continuous
the capability to form immobilized enzyme-sub- process.
strate complex is much lower than that of the
soluble enzyme when large oligosaccharide mol- 4. a-1,6 Enzymes
ecules are used as substrate.29-155
Interest in the cloning and expression of The presence of a-1,6 branches makes the
amyloglucosidase has increased in recent years. production of amylolytic derivatives of starch more
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Saha and Zeikus155 reported that amyloglucosidase difficult than it would be in their absence. As
I from Aspergillus awamori was expressed in mentioned previously, amyloglucosidase can
Saccharomyces cerevisiae by means of the pro- cleave a-1,6 bonds, but rather slowly.51-108 On the
moter and termination region from the yeast eno- other hand, P- and oc-amylase have no activity on
lase gene efficiently secreting amyloglucosidase these bonds.130 Hence, the use of enzymes able to
to the medium. hydrolyze these links quickly is of increasing in-
The amyloglucosidase genes from S. diastat- terest to the modern starch industry.
icus155 and Rhizopus sp.155 have been cloned and Debranching enzymes, such as pullulanase
their nucleotide sequences have been determined. (EC 3.2.1.41) and isoamylase (EC 3.2.1.68), have
The use of the potent inducer cyclopentanone been known for more than a decade,129 but their
resulted in maximum amyloglucosidase concen- use has not gained widespread acceptance. In re-
tration in a recombinant A. nidulans.103 cent years, the use of these enzymes has attracted
The maltogenic exoamylase P-amylase (EC more attention as the efficiency of the hydrolysis
3.2.1.2), of plant origin has been known for reaction could obviously be improved by incor-
many years. 30 Plant sources such as barley, porating a specific debranching enzyme in the
wheat, sweet potatoes, and soybeans have been system.
the most common sources of the enzyme.51-66152 Pullulanase was first discovered in 1961154
However, its existence as an extracellular en- and attracted interest because of its specific
zyme in Bacillus sp. and Pseudomonas sp. has action on a-1,6 linkages in pullulan, a linear
been established.66 D-glucose polymer consisting essentially of
P-Amylase acts in an exo-fashion, from the maltotriosyl units joined by a-1,6 bonds (Fig-
nonreducing ends of the outer chains of starch or ure 3),130 starch, amylopectin, and related oligosac-
related polysaccharides, and proceeds by gradual charides.154 Pullulanase alone liberates products
removal of maltose units in the |3-anomeric form of average DP, varying from 115 to 2300 after
(Figure 2). Amy lose is converted almost com- hydrolysis of starch.169 This enzyme is generally
pletely to maltose, whereas amylopectin and other used in combination with amyloglucosidase, <x-
branched polymers are hydrolyzed to various or p-amylase.51-129-130-154-169
extents depending on the degree of branching;148 Pullulanase is produced by mesophilic organ-
this hydrolysis results in about 50 to 60% conver- isms like Klebsiella aerogenes (Table I). 154 The
sion to maltose. This is due to the inability of optimum temperature is around 60°C; this is very
P-amylase to hydrolyze a-1,6 linkages. The high- important because in the glucose syrups industry,
est activity ocurrs at pH 5.0. The temperature of saccharification is normally carried out at this
optimum activity ranges between 55 and 60°C temperature. 51 The use of pullulanase with
(Table 1). amyloglucosidase simultaneously during saccha-
381
is the conversion of glucose to fructose by the
enzyme glucose isomerase (EC 5.3.1.5) (Fig-
0-0-1u ure 4).2654,ii5 The first of a series of commercial
L 0
glucose isomerases marketed was a soluble prod-
uct introduced in late 1973.26 It had a high pro-
L 0
duction cost and was used commercially for only
a short time. In 1974, an immobilized form of the
enzyme was introduced.4 Rapid development took
L 0
place and different immobilized isomerases have
been introduced since then.28-42'132-143-188
Cobalt is a well-known activator of glucose
0-0-0-1u isomerase enzymes; on the other hand, basic in-
vestigations have found that glucose isomerases
L 0
from some sources do not require cobalt ions.71-188
Magnesium is another activator of these enzymes
and there is a pH dependence on the required
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382
CH OH
Glucose
i s oMe r a s e
0
HO 1 0H OH
OH
Glucose Fruofcose
ft 1 pha-cuc lodext.r-in
,0-Ov
O
\0
0-0-0-0-0-0-0 n
f
/
°
0 I t Beta-nyc 1 ortex +.r i n
0
0
Starch Molecule .O'°-O
\
X
— cycl o d e x t r i n
sist of D-glucose units linked primarily by a-1,4 stance) can reduce the same amount of Cu2+ in
bonds having a DE <20.137 The term 20 DE Fehling solution to Cu+ as 20 parts of pure
signifies that 100 parts of the syrup (dry sub- D-glucose.23-36
383
Maltodextrins are prepared as a white powder
STARCH
or concentrated solutions by partial hydrolysis of
SUSPENSION
starch with safe and suitable enzymes. Malto-
(30-4B V. « / u ; PH 6.5;
dextrins are generally recognized as safe (GRAS)
; alpha-attylase)
as a direct human food ingredient at levels consis-
tent with current good manufacturing practices. 63
On the other hand, corn syrup solids are defined \
by the FDA as dried glucose syrups in which the GELATINIZftTION
STEAM
reducing sugar content is 20 DE or higher. 137 ( 1 1 8 - 1 4 8 °C; 5 - 1 9 win)
Traditionally, maize starch has been used for
the manufacturing of these products. The work on
maize-based products has been continued in order
LIQUEFACTION
to extend the knowledge as to how a good starch
( A l p h a - a n y l a s e ; 98-95 °C;
product can be made from this important raw
90-120 Min)
material,11-21 but some attention has also gradu-
ally been given to the production of maltodextrins
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B. Manufacturing Processes
FILTRATION AND
CARBON PURIFICATION
Maltodextrins and corn syrup solids are com-
monly prepared from regular and waxy corn starch8
by controlled enzymatic hydrolysis called lique-
faction (Figure 6).51-113-150-165 A starch slurry is
adjusted to the desired dry solid concentration (30
to 40% dry solid basis),7-165 pH adjusted at 6.5
with acid and then the enzyme (B. licheniformis
cc-amylase) and calcium ion are added. The starch
slurry is pumped through a jet-cooker where the
temperature is increased to 140°C and held for 5
to 10 min.148-165 Exiting the jet-cooker, the gelati- HALTODEXTRINS
nous starch is discharged into a liquefaction reac- OR
tor. At this point, the dispersed starch polymer COB! S¥KU? SOLIDS
material becomes more accessible to the en-
zymes.109 Then the gelatinized starch is vented to
the atmosphere and cooled to the liquefaction
temperature (90 to 95°C).148 A further addition of FIGURE 6. Liquefaction of starch to produce malto-
dextrins, and corn syrup solids. (Adapted from Refer-
bacterial oc-amylase takes place at this stage be- ences 51, 62, 104, and 107.)
cause the initial quantity of enzyme added has
been destroyed by high-temperture treatment.113
The liquefaction is allowed to continue until solids.51-69 Then, the pH value is decreased to 3.0
the required DE value is reached. Usually after to 5.0 and the slurry is boiled for about 5 min to
about 1 h, the DE is 10 to 12, and by 2 h it is 15 inactivate the enzyme.126 Color in the hydrolyzate
to 16.51113I5° By controlling the reaction time and/ is removed in granulated carbon columns. These
or by adjusting the enzyme dosage, it is possible columns are renewed by the removal of spent
to obtain a DE from 20 to 39 for corn syrup carbon and the addition of regenerated carbon on
384
a regular basis.63-106 The carbon pulsed out of the sold in concentated solution form; there are fac-
column is washed with water to remove the ad- tors affecting the storage stability of these prepa-
hering syrup, and the solution is returned to the rations, that is, the stability of 50% (w/w) aque-
feed-line of the carbon column. The washed car- ous solution of low DE maltodextrins is frequently
bon is revivified by heating it in a gas-fired fur- poor, with precipitation occurring at 25°C.85
nace at approximately 920°C in an atmosphere Some of the most useful functional properties
carefully controlled to maintain a low degree of are low hygroscopicity, bland flavor, extremely
oxidation.109 low sweetness, high viscosity, and the ability to
The resulting syrup is then evaporated, spray retard ice crystal growth in ice creams, ice milk,
dried, and packaged. and other frozen novelties.8'63'107 They contribute
Acid liquefaction requires the use of corro- chewiness, binding properties, and viscosity in
sion-resistant materials and gives rise to high-salt soft candy, and increase the shelf life and main-
ash contents.165 Originally, batch processing was tain moisture levels of hard candies.63 Malto-
used in which a starch slurry was adjusted to a pH dextrins also have application in pan coatings and
of 1.8 to 2.0 with hydrochloric acid. The acidified tableting,39 or can be used as partial replacements
starch was heated at about 90°C for 30 min and for albumen in marshmallow formulation based
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385
Starch Haltodextrins Corn syrup
solids
Dextrose equivalent 5 18 15 18 25 36
Binding
Bodying agent
Browning reaction
Cohesiveness
Flavor enhancement
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Hygroscopicity
Solubility
Sweetness
Viscosity
other firms, says the company, because it recog- without fat is difficult and many may have se-
nizes the properties and roles of different fats in rious flavor defects. Some even suggest that
foods and then tailors the starch to have the orga- small amounts of fat, qualifying for low-fat
noleptic and functional properties needed for spe- rather than fat-free standards, may creep back
cific applications. into foods to compete with better tasting prod-
Other products contain proteins and surfac- UCtS.79'96-98'173
tants in addition to low-DE dextrins, which are
designed to replace up to 81% fat in cakes.96
Formulation of low-fat foods requires reformu- 2. Corn Syrup Solids
lation of traditional products, sometimes with
different ingredients. When fat replacers are Corn syrup solids (DE > 20),107 like maito-
used, the carbohydrate molecule is hydrated to dextrins, as individual products differ in then-
achieve fat-mimicking functionality. Addition- degree of hydrolysis, but in general present the
ally, sensory attributes such as the degree of following typical carbohydrate profile: 0.8 to
cohesiveness, firmness, dryness, and juiciness 16.9% glucose (DP = 1), 5.5 to 26.9% maltose
must also be considered.184 Making products (DP = 2), 9.1 to 16.4% maltotriose (DP = 3), 6.8
386
to 10.0% maltotetraose (DP = 4), and the rest is VI. SWEETENERS
higher saccharides (DP > 4) (40.4 to 77.8%).51-63
Corn syrup solids are granular, crystalline, A. Introduction
or powdered forms of corn syrup.63 They pro-
vide a protective coating and sheen in panned The development of corn sweeteners can be
goods, and plasticity and viscosity in chewy con- traced back to the early nineteenth century, when
fections.39 They can be used in low-moisture the German chemist Kirchhoff discovered that
products, provide the ability to increase solids starch yielded a sweet substance when heated
without cooking, and make blending with dry with acid.109-161 Nevertheless, the ancient Chi-
ingredients easier.8-107 Corn syrup solids are good nese 1000 years before Christ made "Hung Fan,"
materials for flavor encapsulation and oxygen meaning "sweet from cereal."30 Kirchhoff s dis-
uptake, that is, oxidation of orange oil decreases covery was not used industrially on a large
as DE increases.149 Because there is a strong scale until 1850, and since then, the manufac-
dependence on oxidation on DE of the hydro- ture of glucose and other syrups through acid
lyzed starch,38'120-149 it has been observed that a hydrolysis has expanded considerably.51 In
product with the highest DE (36.5) would be 1815, de Saussure identified acid hydrolysis as
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extremely stable and would have a shelf life of the underlying reaction of Kirchhoff s observa-
years, without using an antioxidant. tion. He found that the end product of the hy-
drolytic reaction was dextrose.106
The discovery, isolation, and application of
V. OLIGOSACCHARIDES various carbohydrase enzymes resulted in the
development of many new corn syrups.51-108 In
Starch oligosaccharides are composed of the early 1960s, the first commercially signifi-
cc-D-glucopyranosyl units linked by a-1,4 and/or cant quantities of crystalline dextrose were be-
a-1,6 bonds. The generic term "oligosaccharide" ing marketed. It was produced from corn starch
is customarily used for saccharides having a DP by an all-enzyme process using bacterial a-
of 2 to 10. The large-scale production of malto- amylase for starch liquefaction and fungal
oligosaccharides larger than maltotriose has not amyloglucosidase for saccharification of the
been fully developed because these compounds hydrolyzate to produce an approximately 95%
are rather difficult to manufacture. Oligosaccha- dextrose level.68-106
rides with a DP > 3 have been considered only as Hydrolysis with the P-amylase of barley malt
research chemicals and for medical use.133 was a process established many years ago.51-127 In
The discovery of the microbial enzymes that 1940, a new class of corn syrup sweeteners that
produce specific oligosaccharides, such as were clear, colorless, and exceptionally sweet
maltotriose, maltotetraose, maltopentaose, and appeared on the market. These new syrups were
maltohexaose, has made posible the manufacture made with the bacterial enzyme P-amylase and
of syrups with a high content of malto- amyloglucosidase, which cause the production of
oligosaccharide. These syrups are produced from D-glucose and the concomitant formation of mal-
liquefied starch (DE 3) using the specific enzyme tose from oligosaccharides.109 The syrups imme-
in combination with debranching enzymes, such diately enjoyed great demand because of their
as pullulanase or isoamylase.133 Such syrups are enhanced sweetness.
new products having low sweetness. They pro- The commercial development of glucose
vide resistance to retrogradation of starch gels isomerase, which converts glucose to its sweeter
and prevent crystallization of sucrose and are now isomer fructose,26 was a major milestone in the
being used as enhancers for various foods, pow- corn sweetener industry. The first high-fruc-
dering materials, saccharides for dry milk, and tose corn syrup, commonly referred to as HFCS,
liquid diets for patients. High purity malto- appeared in 1967 and contained 15% fruc-
oligosaccharides are also used as substrates for tose.68'104 Further process improvements resulted
measuring oc-amylase levels in human serum and in HFCS products containing 42 and 55% fruc-
urine in clinical tests.133 tose.24-68-106
387
B. Dextrose Syrups tion is the hydrolysis of oligosaccharides or dex-
trins to low-molecular-weight sugars such as glu-
1. Manufacturing Processes cose, maltose, or a mixture of these and their by-
products.109
The manufacturing of dextrose feedstocks The starch slurry, pH 6.5, is gelatinized by
from starch is a multistage process involving ther- cooking at high temperature with cc-amylase. The
mostable cc-amylase for liquefaction and gelatinized starch is then liquefied by thermo-
amyloglucosidase for saccharification in succes- stable a-amylase. The product of this reaction is
sive enzymatic steps (Figure 8).137 Saccharifica- a soluble dextrin hydrolyzate with a DE of 5 to
15 5i,i29 After liquefaction, the pH of the slurry is
adjusted to 4.5 and the temperature lowered to
STARCH 60°C. Amyloglucosidase is now added and the
SUSPENSION
saccharification reaction is allowed to continue
(38-48 •/. u/v; pH £.5;
for 48 to 96 h in a stirred tank until the maximum
&**; alpha-anylase)
level of glucose is reached.106154'165
388
and color content.137-165 Consequently, the extent must be operated at the highest possible substrate
and expense of refining are increased. concentration, the lowest possible enzyme con-
If pullulanaseor isoamylase are used simulta- centration, and the lowest possible residence
neously with amyloglucosidase to saccharify a time.88
partially hydrolyzed starch, a slight increase in
glucose yield can be obtained. Less saccharifica- 2. Functional Properties and
tion time and amyloglucosidase dosage are re- Applications
quired; therefore, less isomaltose formation due
to reversion reactions takes place and the maxi- The carbohydrate composition of typical glu-
mum glucose levels achieved are higher.129-130-154 cose syrups are as follows: 94 to 98% glucose, 1
The debranching enzyme takes care of the branch to 3% maltose, 0.3 to 0.5% maltotriose, and 1 to
points in the amylopectin so that the amylo- 2% higher saccharides.106-109 Glucose syrups are
glucosidase has to hydrolyze only the linear oli- used for nonyeast fermentation purposes (phar-
gosaccharides.13°-154 maceuticals, organic acids, or vitamins),51 for the
Replacing soluble amyloglucosidase with its manufacture of crystalline D-glucose,109 or in the
immobilized form allows for using the same en- production of fructose syrups by further continu-
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389
extremely high maltose syrups, and (3) high con- enzymes cannot act on a-1,6 linkages, the final
version syrups.153 The general DE value of these maltose content is limited to around 60%.148 As
syrups and their saccharide composition is given the reaction is self-terminating, no specific en-
in Table 2. Starch hydrolyzates of 35 to 60% zyme inactivation step is required.109
maltose content on a dry substance basis, after In the case of extremely high maltose syrup
refining and concentration, are the so-called high production, basically the same steps for the pro-
maltose syrups.126 Extremely high maltose syrups duction of high-maltose syrups are followed (Fig-
have a 70 to 85% maltose content.125 High con- ure 9). However, one of the most important
version syrups are characterized by their high diferences is that it is necessary to use a
glucose (35 to 43%) and maltose (30 to 47%) debranching enzyme, such as pullulanase or
contents; they have the highest DE (60 to 70).156 isoamylase, in addition to P-amylase or fungal oc-
These syrups have a high fermentable sugar con- amylase.125'126-129 Incorporating debranching en-
tent (>85%).152-153 zymes into the saccharification system produces
higher maltose levels (Table 2).
The high-conversion syrups are produced by
2. Manufacturing Process saccharifying a liquefied starch by using both
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TABLE 2
The Composition of Maltose Syrups and Their Dextrose Equivalent (DE)
390
STARCH
SUSPENSION
(38-40 X. w/v; pH £.5;
Ca++; alpha-aitylase)
J
GELATINIZATION
STEAM
(110-148 °C; 5-10 * i n )
LIQUEFACTION LIQUEFACTION
CE 5-18 1>E 38-48
(fllpha-amjlase; 98-95 °C; (flcid/alpha-aiwlase)
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90-128 Nin>
SACCHARIFICATIOH
SACCHARIFICATION SACCHARIFICATION BE 68-70
BE 38-35 BE 45-68 (Fungal alpba-aHylase or b e t a -
(Fungal alplia-awlase or beta- (Fungal alpha-anylase o r b e t a - aitylase; anuloglucosidase;
a n g l a s e ; ?H 5 . 8 - 5 . 5 ; 58-55 °C> anylase; pullulanase o r pH 6 . 8 ; 75 °C)
i s o a x y l a s e ; pH 6 . 0 ; 50-55 °C)
<
ENZYME INACTIUATION
FILTRATION FILTRATION (80-85 aC; 15 n i n )
AND AND
PURIFICATION PURIFICATION
FILTRATION
am
PURIFICATION
FIGURE 9. Production of various maltose syrups by conventional processes. (Adapted from References 51,149,
and 150.)
The clarified solution may then be concentrated use and recovery, low cost and convenience of a
by evaporation. continuous reaction system, and higher reaction
The development of immobilized enzyme rates due to high concentration of enzymes. Im-
systems for maltose syrup production is advanta- mobilization of P-amylase on a variety of carriers
geous for several reasons, including enzyme re- is documented.35 Although immobilized enzymes
391
are not yet used in industry for starch hydroly- baking, confectionery, canning, soft drinks, etc.156
sis,152 some attempts have been made to develop Although these syrups have a high glucose con-
a continuous maltose production process under tent (35 to 42%), they are stable enough to crys-
industrial-scale conditions.35 Making use of the tallization at temperatures as low as 4°C and at 80
appropriate carrier and enzyme and applying the to 83% dry substance.152
correct immobilization procedure, a conjugate
with acceptable initial performance and stability D. High-Fructose Corn Syrups
can be obtained.
Continuous production of high maltose syr- 1. Introduction
ups in an ultrafiltration reactor has been devel-
oped by simultaneously using P-amylase and Beet sugar usage grew along with that of
isoamylase.77 dextrose and corn syrup, but other sources of
nutritive sweeteners were still sought. Sucrose
3. Functional Properties and was known to be a disaccharide consisting of
Applications 1 mol of D-glucose and 1 mol of D-fructose, linked
by an oxygen bridge at the respective aldose and
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In general, various maltose syrups are used in ketose functions. Fructose is also found in the free
the brewing, baking, soft drink, canning, and con- state in certain fruits, vegetables, and honey.
fectionery industries. These syrups can also be As early as 1926, the Hon. Cyrenus Cole,
used as moisture conditioners, stabilizers, carri- U.S. Representative in the House for the State of
ers, and bulking agents.153 They have a decreasing Iowa, advocated growing the Jerusalem artichoke
tendency to crystallize and are relatively (Helianthus tuberosus), a variety of sunflower, as
nonhygroscopic. Hence, they are used in the manu- a crop because its tuber was rich in the polymer
facture of hard candies by the confectionery in- inulin, which could be hydrolyzed to levulose.17
dustry and in frozen desert formulations, where This term was used in preference to fructose to
they control ice crystal formation.51 designate the industrial product for the next 50
In particular, the high maltose syrups are char- years. Levulose remained a target, and an elusive
acterized by their low viscosity of solution, low one, until the commercialization of glucose
hygroscopicity, good heat stability, and mild isomerase in the late 1960s and early 1970s.27
sweetness.126 They may also be used as nutrients Then the name was changed and attention turned
for children,125 or to impart reduced browning to the economic potential of fructose.
capacity.153 The first shipment of HFCS was made in
Extremely high maltose syrups are used to 1968, and in 1985 shipments exceeded 10 billion
produce pure crystalline maltose, which is of in- lbs of dry solids.104 This shipment of HFCS was
creasing interest in the pharmaceutical industry, the first generation of fructose syrups, which con-
where it may be used instead of D-glucose for tained 42% fructose and 50 to 55% dextrose.24-106
intravenous feeding of diabetics.126152 Crystalline The syrup served as only a partial replacement for
maltose would meet the requirements of those sucrose. The use of HFCS in beverages did not
applications due to its high purity.158 accelerate until the development of the second
The catalytic hydrogenation of maltose gives generation in the mid-1970s. This product con-
rise to maltitol, a polyhydric alcohol, which is a tains 55% fructose and 40% dextrose.99-106 The
low-calorie sweetener and is readily metabo- industry also produces a 90% fructose syrup.
lized.152 The isomerization of maltose produces The use of HFCS in the U.S. has increased
maltulose, which is on the same level as maltitol from 12.1 lb/person in 1978 to 42.0 lb/person in
in sweetness properties; it is 75% as sweet as 1986.45 By 1990, the per capita intake of corn
sucrose, resistant to crystallization, and useful as sweeteners was 61.3 lb;53 a high percentage of
a humectant. Chewy candies, fondant, and hard this was HFCS. This per capita increase has been
candies are examples of its applications.39 supported by one of its most desirable attributes:
The high conversion syrups have wide appli- sweetness. It has been reported that fructose has
cations in many industries, for example, brewing, a sweetness of up to 1.8 times that of sucrose.84-163
392
2. Manufacturing Processes In the refining process, the high-dextrose
conversion liquor is filtered on rotary precoat
Preparation of very high-quality dextrose feed- filters to remove coagulated protein and oil (Fig-
stock for isomerization is first necessary because ure 10). This filtered liquor is then passed through
of the very low color and ash specifications for a series of check filters to remove traces of par-
the finished HFCS. A high-purity feedstock is ticulate matter.106 First, color is removed, and
also required for efficient utilization of the immo- then the carbon-treated liquor is filtered again and
bilized isomerase enzyme column. passed through an ion-exchange system. It is then
FILTRATION
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CHEMICAL TREATMENT
ISOMERIZATION
glucose isoiterase;
pH 8.5; 55-65 °C; 4h>
GLUCOSE
SECONDARY CARBON TREATMENT,
ION EXCHANGE AND EVAPORATION
CHR0MAT0GR81WC
42ZHFCS FRACTIONATION
I.
BLENDING S8ZHFCS
55ZKFCS
FIGURE 10. Flow chart for the manufacture of 42% high fructose
com syrup as well as 55 and 90% high fructose com syrup.
(Adapted from References 24, 102, and 140.)
393
evaporated to a solid matter of 40 to 45%. Before was developed to produce a 49:51 mixture of
pH is adjusted, the refining glucose syrup is chemi- glucose and fructose from liquefied starch.101
cally treated by the addition of magnesium ions,42
which are activators of glucose isomerase and
also competitively inhibit the action of any re- 3. Functional Properties and
sidual calcium ions, which are potent inhibitors of Applications
the enzyme.188
Byproduct formation at the isomerization One of the most important characteristics
process is a function of temperature, pH, and of HFCS is sweetness. In this respect, 42%
time. Under the conditions used, a residence HFCS is about equal to sucrose (Table 3), 55%
time of up to 4 h is considered safe, and the HFCS is slightly higher, and 90% HFCS has
small amount of color formed can be removed the greatest relative sweetness (180).58 Never-
by conventional carbon treatment after the theless, sweetness values vary with tempera-
isomerization.24'106'132 Use of ion-exchange re- ture, pH, and solids. In general, relative sweet-
fining in addition to carbon is necessary in order ness of fructose is a function of concentration,
to remove ash. temperature, and pH, with the maximum sweet-
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The process used to produce the 55% HFCS ness values being found in cooled, dilute solu-
is also shown in Figure 10. This syrup is made tions at low pH.2484'104 Among other character-
from the 42% syrup by a fractionation and blend- istics of HFCS are the ability to produce food
ing process. It is pumped into the fractionation systems with lower freezing points and higher
unit, where calcium sites retain the fructose while boiling points, thus effectively inhibiting crys-
dextrose and the higher saccharides pass through. tallization,8 and enhancing citrus flavor in fruit
The dextrose and higher saccharide stream is re- products.46-75 There have been some reports
turned to the 42% production channel. Water is about the synergistic effects of HFCS on sweet-
pumped through the fractionation unit to release ness. Increase in sweetness is perceived when
the fructose. The outlet stream generally is 80 to fructose is used with other nutritive and
90% fructose. It is then blended with the 42% nonnutritive sweeteners.24-75
HFCS to make the 55% product.2451'104106 The first-generation HFCS, containing 42%
Besides sucrose and starch, other potential fructose, 52% glucose, and 6% of higher sac-
commercial sources of fructose exist. One of these charides (Table 3), found widespread use in
is inulin. Inulin is a linear p-2,1-linked fructose baking and other food products.96 However,
polymer terminated by a sucrose unit residue. It researchers have encountered problems that
occurs as an energy reserve in various plants, limit the frequency and quantity of HFCS in
particularly in those of the Compositae family; baked products.33 Excessive browning of the
good sources are chicory, Jerusalem artichoke,
and dahlia. The enzymatic hydrolysis of inulin, TABLE 3
using inulinase from yeast, has been reported as Typical Composition and Sweetness of High
an alternative way to produce fructose.187 There Fructose Corn Syurps (HFCS)
have been other attempts to produce fructose from
other starch sources like cassava16-56 and broken 42% 55% 90%
rice.31 HFCS HFCS HFCS
394
crust and crumb of cakes containing HFCS was decreases in aqueous solution to about 160.58
attributed to the Maillard-browning reactivity One of its applications is in food for diabetics39
of monosaccharides in the HFCS.114181 Johnson because it is metabolized without insulin.
et al.80 controlled the browning of cakes pre-
pared with up to 100% HFCS as a sucrose
replacement with the acidulant potassium acid VII. CYCLODEXTRINS
tartrate or cream of tartar. The total browning
and 5-hydroxymethylfurfural content of the A. Introduction
crust and crumb was altered as the amount of
HFCS replacement increased, indicating that Cyclodextrins have been known for nearly
the acidulant reduces the browning reactivity 100 years. In 1891, Villiers discovered crystalline
of monosaccharides by lowering the pH of the dextrins, "cellulosines", in bacterial digests of
batter prior to baking. The replacement of su- starch; as early as 1903, Schardinger demonstrated
crose by other sweeteners in cakes is not as the cyclic structure of these compounds.72-160 Be-
simple as it is in breads and requires special cause of Schardinger's studies, cyclodextrins were
reformulation of the system. The effect observed initially referred to as Schardinger's dextrins; of
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in cakes when sucrose has been replaced by more significance was the fact that his work, al-
42% HFCS includes a decrease in baking time, though crude, set the direction for future research,
lower cake volume, change in taste and sweet- pointing it toward a study of the structure of
ness, and gummy mouthfeel.96 cyclodextrins and their preparation. In 1938,
The second generation HFCS, containing Freudenberg confirmed the ringed structure of
55% fructose, 40% glucose, and 5% higher sac- cyclodextrins and their ability to form inclusion
charides, has major use in beverages, especially complexes.146
in soft drinks. Cola drinks account for approxi- Three main types of cyclodextrins are known
mately 63% of the soft drink volume. In 1980, (Figure 5): a-, P-, and y-cyclodextrins consisting
the 55% HFCS was first used in a major cola at of six, seven, and eight glucose monomers, re-
a 50% sweetener replacement level, and in 1984, spectively.72 These monomers are bound together
all cola manufacturers approved usage of the in a doughnut-shaped ring, giving the cyclodextrins
syrup at a 100% replacement level. In the late a molecular structure that is relatively rigid and
1970s and early 1980s, the noncola soft drink has a hollow cavity of a specific volume. These
producers switched to 100% replacement with cyclic molecules have neither a nonreducing nor
H F C S 68,104 reducing end-group.
The 55% HFCS was also used as replace-
ment for sucrose in sugar-snap cookies. It was
reported that 55% HFCS satisfactorily substi- B. Manufacturing Processes
tuted for 10 or 30% of the sucrose in this type
of cookie.10 The cookies appear to dry out as For commercial production of cyclodextrins,
they cool because the water yielded by the su- two basic processes are used: the solvent and the
crose, as it crystallizes, is bound by the flour nonsolvent methods.72-176 In the solvent method, a
components (starch and protein). HFCS acts as solvent directs the enzyme reaction to produce
a crystallization inhibitor and will keep sucrose predominantly one cyclodextrin. Using this
in solution, producing a cookie that retains its method, the reactor is filled with the starch sub-
soft texture.59-96-181 strate, the enzyme, and the solvent. The substrate
A very small amount of 90% HFCS is sold is generally a partially hydrolyzed starch with a
commercially.104 It is available in liquid and DE <10.
crystalline forms; the latter is a relatively new At the end of the reaction period, the reactor
product developed from advanced technology.75 contains a mixture of an insoluble cyclodextrin
Crystalline fructose, the P-pyranose form, has complexed with the solvent (i.e., oc-cyclodextrin
the greatest relative sweetness (180), but this forms an insoluble complex with 1-decanol, and
395
(5-cyclodextrin precipitates out with cyclo- form inclusion complexes with a wide variety of
octane),128 starch hydrolysate produced by the organic molecules of suitable size (guest mol-
transglycosylation and hydrolysis reactions, and ecules).
the residual chemical. The cyclodextrin complex In aqueous solution, the slightly apolar
is separated from the other components by cen- cyclodextrin cavity is occupied by water mol-
trifugation or filtration. After removal of the ecules. This polar-apolar interaction is energeti-
chemical from cyclodextrin, the soluble cyclo- cally unfavorable and water can be readily dis-
dextrin is refined like any other soluble starch- placed by appropriate guest molecules that are
derived product and crystallized. The cystals less polar, provided that their geometrical dimen-
are then recovered and dried. Because the sepa- sions match those of the cavity.146-174 This inclu-
ration processes are highly efficient, the level of sion complex formation is also known as "mo-
residual chemical in the final product is <1 ppm.72 lecular encapsulation". The inclusion complexes
In the nonsolvent process, the reactor is can be isolated as stable crystalline substances.
filled with a starch hydrolysate and the en- The most important parameter for complex
zyme. As no organic chemicals are present to formation with hydrophobic substances is their
prevent the growth of microorganisms, which three-dimensional size (Table 4). Thus, for ex-
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could lead to contamination of the final product ample, oc-cyclodextrins typically form inclusion
with microbial metabolites, aseptic or sterile complexes with aliphatic hydrocarbons, but, curi-
conditions must be used. At the completion of ously, also with gases like ethylene and carbon
the reaction, the CDGTase is inactivated by dioxide. (3-Cyclodextrins form complexes with
heat or acid. An oc-amylase unable to hydrolyze molecules such as naphthols or steroids, whereas
the cyclodextrins is added to hydrolyze the y-cyclodextrins can accept bulky guests like vita-
noncyclic dextrins.72 min D2.160
After refining, water is evaporated from the Although cyclodextrins have been studied for
reaction mixture to maximize the yield of the nearly a century, they were not utilized for food
(3-cyclodextrins, which is the least soluble and applications until the last decade for several rea-
generally the most readily crystallizable cyclo- sons. First, cyclodextrins were expensive to pro-
dextrin. If too much water is removed, more duce and available in only limited amounts; sec-
than one cyclodextrin will be crystallized. The ond, incomplete studies reported them to be toxic;
crystals are collected by centrifugation or fil- and third, there was not enough knowledge to
tration, washed with a small amount of water, produce them on an industrial scale.146-160 How-
and dried. ever, because of continuing research, these barri-
ers have been gradually removed so that use is
C. Functional Properties and now technologically possible in a variety of food
Applications applications.
Perhaps the greatest potential for cyclodextrins
The attraction of these cyclic, torus-shaped is in the area of flavors and spices. Flavor con-
maltooligosaccharides arises from their ability to stituents often have an unstable nature, which
TABLE 4
Properties of Cyclodextrins
396
may restrict their application into formulated foods to be a success, especially for fructose syrup pro-
and shorten the shelf life of the product. duction. However, in order to improve the
Cyclodextrins, however, can stabilize volatile economy of enzymatic catalysis in the production
materials.146 of maltodextrins, glucose, or maltose, highly effi-
Cyclodextrins can also stabilize emulsions of cient processes have been developed. Today,
fats and oils, shielding them from oxidation and specific amylases allow batch preparation of tai-
thus preventing rancidity.175 They can also be lor-made specific maltodextrin products, cyclo-
used to debitter citrus juices. This application is dextrins, or high glucose and maltose syrups.
extremely important because, previously, the cit- Maltodextrins are important components of
rus industries have tried to mask the bitterness in foods, contributing a broad range of functionalities.
juice blends rather than remove it.94-164 One of these is as a fat substitute. Financial ana-
With respect to bioconversions and fermenta- lysts forecast that fat substitutes may match or
tions, cyclodextrins apparently possess an advan- even surpass the success of sugar substitutes.57 In
tageous combination of properties. First, they 1991, two thirds of the adult population, 124
enhance solubilization of organic compounds. million Americans, were using products with low
Second, they reduce toxicity. Complexation of or reduced fat. These consumers included 72%
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organic substrates and/or products may signifi- women and 61% men.121 There are now two dis-
cantly reduce their concentration.13 Third, they tinct types of light food consumers: (1) dieters
are biocompatible. Cyclodextrins are known to and (2) calorie-, fat-, and health-conscious
cause no damage either to free enzymes or to nondieters. These consumers represent signifi-
microbes.13 cant markets.
Although the potential of cyclodextrins is As the demand for starch-based sweeteners
well documented, the market for these products grows, high-maltose- and high-fructose-contain-
is still limited. Two important facts are respon- ing syrups will gain importance, principally as
sible for the present market situation: their high industrial commodities for use as food ingredi-
cost and the limited availability of a- and y- ents. The production of maltose-containing syr-
cyclodextrins.160 Cloning and overexpression of ups has increased for the last several years in
CGTase genes provide a straightforward way to Japan and the U.S. An increasing interest in mal-
satisfy the anticipated expansion of the tose products is also predicted in the European
cyclodextrin market.160 markets.158 The market for high-fructose syrups
A novel production of cyclodextrins in the has grown dramatically in the United States espe-
tubers of transgenic potato plants has been devel- cially for the soft drink industry.
oped.131 This report declares that even though the The various applications for cyclodextrins
levels of cyclodextrins were low, this is the first include pharmaceutical delivery systems, flavor
step in using genetic engineering for the "molecu- and odor enhancement, and the removal of un-
lar farming" of novel carbohydrates. desirable compounds (such as caffeine) from
foods. In view of their high potential, as
cyclodextrins prices continue to drop, there will
VIII. CONCLUSIONS undoubtedly be much research in this area. It is
necessary that the fine structure analysis of the
Food processing through enzyme utilization enzyme CGTase be completed. Then it may be
offers many advantages. Starch transformation is possible to engineer more specific a- or y-CGTases
an industry in which chemical processing has or even CGTases that catalyze the synthesis of
yielded almost completely to enzyme processing. cyclodextrins with more than eight glucose resi-
Amylolytic enzymes have gained importance be- dues.
cause of their ability to catalyze batch reactions As the food industry continues to move to-
under moderate conditions, which are not reached ward developing new amylolytic enzymes and
with chemical processes wherein nonspecific hy- novel starch processes to provide products that
drolysis and formation of byproducts are present. meet consumers' high expectations, sales will
The technique of enzyme immobilization proved continue to increase.
397
ACKNOWLEDGMENT 16. Berghofer, E. and Sarhaddar, S., Production of glu-
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This work was supported by Consejo Nacional 188, 1988.
de Ciencia y Tecnologia (CONACYT). 17. Bernetti, R., From corn syrup to fructose, Cereal
Foods World, 35, 390, 1990.
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