Food Bioprocess Technol (2014) 7:610–618
DOI 10.1007/s11947-013-1144-2
COMMUNICATION
Optimization of Enzymatic Production Process of Oat Milk
Using Response Surface Methodology
Aastha Deswal & Navneet Singh Deora &
Hari Niwas Mishra
Received: 10 December 2012 / Accepted: 5 June 2013 / Published online: 7 July 2013
# Springer Science+Business Media New York 2013
Abstract The simultaneous effects of slurry concentration
(25–35 % w/w), enzyme concentration (0.5–2.5 % w/w) and
liquefaction time (30–90 min) on the yield, total solids and
rheological parameters of oat milk developed were studied
and optimised by the application of response surface meth-
odology. The effect of independent and dependent variables
have been studied using a central composite rotatable design
of experiments. Power law model explains the flow behav-
iour of the developed oat milk samples with correlation
coefficient (R2) varying from 0.89 to 0.96. All the formula-
tions exhibited pseudo plastic behaviour with the flow be-
haviour index (n) between 0.29 and 0.46 and consistency
index varying from 1.033 to 10.22 Pa sn. Statistical analysis
showed that yield, total solids and rheology of oat milk were
significantly (p<0.05) correlated to slurry concentration,
enzyme concentration and liquefaction time. The optimum
conditions for making oat milk were: 27.1 % w/w slurry
concentration, 2.1 % w/w enzyme concentration and lique-
faction time of 49 min.
Keywords Oat milk . Enzymatic liquefaction . Rheology .
Response surface methodology
Introduction
In recent years, health concerns and changes in consumer’s
preferences towards functional foods have led to an extensive
research on developing healthy foods. There has been an
increased demand from consumers for functional drinks with
high acceptance and functionality. Soy milk has been the first
non-dairy functional drink in the market. It is a very popular
A. Deswal (*) : N. S. Deora : H. N. Mishra
Agricultural & Food Engineering Department, Indian Institute
of Technology, Kharagpur 721302, West Bengal, India
e-mail: deswalad@gmail.com
beverage amongst Asians and is gaining popularity globally as
well. Soymilk is a rich source of protein, isoflavones, saponins
and fibre (Bricarello et al. 2004). The increasing consumption
of soy milk has led to the need of developing functional drinks
from other sources too. Oats are one of the promising raw
materials for developing health beverage.
Oat belongs to the genus Avena and is considered a minor
cereal crop based on annual production. It is primarily used
as an animal feed, but is gaining popularity as a breakfast
cereal in forms like oat meal, ready-to-eat cereal and cereal
bars. Oat products are whole grain and heart-healthy. Oats
(Avena sativa L.) have good nutritional profile and proven
health benefits. Oats are a good source of protein, fat and soluble
fibre, β-glucans which can act as fat replacer and stabiliser. The
protein content of oat typically ranges from 15 to 20 % and
globulins are the major proteins of oats (70–80 %). Oat protein is
considered to be more nutritious than most cereals consumed
today (Robert et al. 1983; Zarkadas et al. 1995).
Oats are known to reduce blood cholesterol levels as well as
glucose and insulin responses (Wood et al. 1994; Kirby et al.
1981; Hallfrisch et al. 1995; Anderson et al. 1990; Tiwari et al.
2012). Braaten et al. (1994) showed that isolated oat β-glucan
when mixed in a drink significantly contributes in reducing the
total and LDL cholesterol levels of hypocholesterolemic adults
but did not alter the level of HDL cholesterol (Braaten et al.
1994). Oat β-glucan is a linear polysaccharide that consists
only of β-D-glucopyranosyl units joined by either (1→3) or
(1→4)-β-D-linkages (Morgan et al. 2000; Sharma and Gujral
2012; Tiwari and Cummins 2010). β-Glucan content in oats is
estimated to be around 3.9–7.5 g/ 100 g of dry mass
(Gajdošová et al. 2007) . A hydrolysis product of oat starch
known as oat dextrin (OD) with a dextrose equivalent (DE)
value less than 20 possesses different physicochemical prop-
erties including solubility and viscosity (Jun-Liang et al.
2008) which are widely used in food industry as a fat substi-
tute, such as in dairy products (Liu and Wang 2006). Oat is
also a source of many compounds that exhibit antioxidant
Food Bioprocess Technol (2014) 7:610–618 611

Table 1 Decoded values of independent variables used in the experi- The process used can be either chemical or enzymatic. Acid
mental design
hydrolysis is widely used in the starch industry to prepare thin
Variables Coded Range of levels boiling starches for use in food, paper, textile and other
symbol industries (Rohwer and Klem 1984). The typical procedure
−1.682 −1 0 1 1.682 to manufacture acid-thinned starch involves treating a concen-
Slurry concentration (w/w) A 25 27 30 33 35
trated starch slurry (36–40 % solids) at a temperature below
the gelatinization temperature of the starch (40–60 °C) with
Enzyme concentration (w/w) B 0.5 0.9 1.5 2.1 2.5
mineral acid for a period of time (Wurzburg 1986). When the
Liquefaction time (min) C 30 42.2 60 77.8 90
desired viscosity or degree of conversion is reached, acid is
neutralised and starch is recovered. Hydrolysis can be con-
trolled by the acid concentration, the reaction time and the
activity. Vitamin E (tocols), phytic acid, phenolic compounds temperature. Hoover (2000) reviewed the current knowledge
and avenanthramides are the most abundant antioxidants in on the mechanism and effects of acid hydrolysis on structures
oat, and flavonoids and sterols are also present (Peterson and physicochemical properties of various starche.
2001; Bratt et al. 2003; Angioloni and Collar 2012). The enzymatic method involves application of liquefying
The major portion of oat consists of starch (50–60 %) enzyme, preferably during gelatinization since it has been
which has a gelatinization temperature in the range of 44.7 found that when starch is gelatinised the semi-crystalline nature
to 73.7 °C (Tester and Karkalas 1996) posing a problem of granules becomes totally amorphous and the starch becomes
during the heat processing of oat milk as the liquid milk sets digestible by amylases (Tester et al. 2006). The enzymatic
into a gel at high temperature. Hence, the purpose of the work liquefaction process has been shown to increase in yield of
is to optimise a process for making oat milk which maintains jicama (Pachyrhizus erosus) tuberous root (Ramos-de-la-Peña
its liquid state at high temperature. The process used should et al. 2012), saccharification of corn starch (Carr et al. 1982),
employ hydrolysis of starch, converting starch into malto- increase in yield of apple juice (Will et al. 2000), recovery of a
dextrins so that no gelatinization occurs at high temperature. carotene-rich functional food ingredient from carrot pomace

Table 2 Matrix of experimental

central composite rotatable de- Experiment Factors Response variables
sign (CCRD) for enzymatic pro- no.
duction of oat milk Slurry Enzyme Liquefaction Consistencya Yielda Totala
concentration concentration time (min) index (Pa sn) (wt%) solids (%)
(wt%) (wt%)

1 30.0 0.5 60.0 8.35±0.03 56.01±0.02 28.90±0.01

2 30.0 2.5 60.0 1.40±0.02 72.31±0.03 27.99±0.01
3 27.0 2.1 42.2 1.74±0.03 75.57±0.03 25.51±0.04
4 30.0 1.5 60.0 5.12±0.01 61.34±0.02 28.14±0.03
5 30.0 1.5 90.0 2.98±0.04 66.54±0.04 27.89±0.06
6 30.0 1.5 30.0 6.78±0.03 55.37±0.05 28.92±0.02
7 30.0 1.5 60.0 5.94±0.05 64.01±0.09 28.28±0.04
8 30.0 1.5 60.0 4.32±0.01 63.43±0.05 27.98±0.04
9 27.0 0.9 42.2 9.83±0.02 56.32±0.03 26.21±0.03
10 30.0 1.5 60.0 5.34±0.03 58.32±0.01 28.73±0.02
11 35.0 1.5 60.0 8.42±0.03 58.87±0.08 27.74±0.02
12 30.0 1.5 60.0 4.99±0.04 65.79±0.02 28.45±0.04
13 30.0 1.5 60.0 4.74±0.01 62.19±0.03 27.57±0.03
14 33.0 2.1 42.2 6.04±0.05 57.41±0.02 28.90±0.02
15 33.0 0.9 42.2 10.22±0.04 53.92±0.07 29.70±0.03
16 33.0 0.9 77.8 7.19±0.04 64.99±0.03 30.30±0.04
17 27.0 2.1 77.8 1.03±0.05 78.87±0.02 24.87±0.05
18 33.0 2.1 77.8 5.09±0.01 56.35±0.01 30.35±0.03
a
19 25.0 1.5 60.0 1.62±0.01 75.08±0.02 22.63±0.05
Mean±standard deviation
20 27.0 0.9 77.8 5.66±0.02 63.58±0.01 25.25±0.01
n=3
612 Food Bioprocess Technol (2014) 7:610–618

Table 3 Estimated coefficients

for the fitted second-order poly- Source Consistency index (Pa sn) Total solids (%) Yield (%)
nomial representing the relation-
ship between the response and Coefficient p value Coefficient p value Coefficient p value
process variables
Constant 48.8740 −70.6936 <0.0001 86.7977
Aa −1.4787 <0.0001 6.8255 <0.0001 −7.0458 <0.0001
Bb −22.3815 <0.0001 −3.3046 0.1329 98.9694 <0.0001
Cc −0.2721 0.0005 −0.3390 0.5435 0.7561 0.0008
AB 0.4543 0.0192 0.0229 0.8401 −2.8058 <0.0001
AC 2.11E−03 0.7051 8.56E−003 0.0429 −0.0013 0.9323
CB 0.0651 0.0377 0.0138 0.4721 −0.1898 0.029
a
A2 0.0201 0.4279 −0.1122 <0.0001 0.1606 0.0367
Slurry concentration
b
B2 0.3549 0.5720 0.4584 0.2928 1.1969 0.4888
Enzyme concentration
c
C2 4.0327E−04 0.5635 4.674E−004 0.3322 −0.0022 0.2563
Liquefaction time

(Stoll et al. 2003) and conversion of corn starch for fat mimic
(Ma et al. 2006). The enzymatic liquefaction of oat starch is
used for the reduction in viscosity and in order to increase the
oat milk yield by facilitating the process of filtration (Aiyer
2005). The process also results in the increase of the amount of
total solids by the action of amylases on starch resulting in
production of maltodextrins (Moore et al. 2005).
The sensorial and rheological properties play an important
role in acceptance of the developed drink by consumers.
Rheological characteristics of food affect the sensory proper-
ties. Also, rheological data are required for calculation in any
process involving fluid flow (e.g. pump sizing, extraction,
filtration, extrusion, purification) and play an important role
in the analyses of flow conditions in food processes such as
pasteurisation, evaporation, drying and aseptic processing
(Marcotte et al. 2001; Rao 2007). Quality characteristics of
oat milk were targeted to provide nutritious, lactose-free,
ready-to-use and consume non-dairy milk substitute.
In this study, enzyme concentration, slurry concentration and
liquefaction time were selected as independent variables for
optimization of the oat milk using α-amylase enzyme. These
were the key factors that influence the activity of the enzyme in
the oat milk. The purposes of the present work are to study the
effect of slurry concentration, enzyme concentration and lique-
faction time on the yield, consistency index and total solid, and
to optimise the enzymatic liquefaction process parameters.

Table 4 Analysis of variance (ANOVA) for the fitted second-order polynomial model and lack of fit for yield, consistency index and total solids as
per CCRD

Response Source Sum of square df Mean square F value calculated p value prob>F

Yield (%) Model 968.3 9 107.59 21.53a <0.0001 Significant

Residual 49.97 10 5
Lack of fit 17.05 5 3.41 0.52a 0.7561 Not significant
Pure error 32.92 5 6.58
Total 1,018.26 19
Consistency index (Pa sn) Model 130.38 9 14.49 21.79a <0.0001 Significant
Residual 6.65 10 0.66
Lack of fit 5.15 5 1.03 3.43a 0.1014 Not significant
Pure error 1.50 5 0.30
Total 137.03 19
Total solids (%) Model 68.22 9 7.58 24.69a <0.0001 Significant
Residual 3.07 10 0.31
Lack of fit 2.28 5 0.46 2.89a 0.1314 Not significant
Pure error 0.79 5 0.16
Total 71.29 10
a
F table (tabulated) 3.46 at p<0.01
Food Bioprocess Technol (2014) 7:610–618 613

Material Methods

Oats used for the study were quick oat groats from the Preparation of Oat Milk
Quaker Oats Co., PepsiCo, India. These are whole oats that
are rolled to flatten them and cut into slightly smaller pieces The processing of oat milk was based on preliminary exper-
so they cook faster. They contain all parts of the oat grain iments. Rolled oats were ground into a laboratory food
including the bran, endosperm and germ portion. Food processor to produce finely granulated oat flour. The ratios
grade α-amylase from Bacillus subtilis was purchased of oat solids and water were taken to form oat slurry
from Sigma Aldrich which had an activity of 489U/mg according to the CCRD design as shown in Table 1.
in a powder form. Calcium chloride at a concentration of 0.04 % (w/w) was

Liquefaction Enzyme
Actual Factor Slurry Conc. =30%
79.0 Actual Factor 73.0
Time = 60 min 76.0 conc. =1.5%
73.3 70.8 68.0

67.5 63.0

Yield
65.5
Yield

Yield

61.8 60.3 58.0

56.0 55.0 53.0

2.1 33.0 77.8 33.0 77.8 2.1

1.8 31.5 68.9 31.5 68.9 1.8

1.5 30.0 60.0 30.0 60.0 1.5

51.1 1.2 B: Enzyme Conc.

B: Enzyme Conc. 1.2 28.5 A: Slurry Conc. 51.1
C: Liquefaction time 28.5 A: Slurry Conc. C: Liquefaction time
0.9 27.0 42.2 27.0 42.2 0.9

(a)

29.6 29.7

27.9 27.9 Slurry Conc. =30%

Total Solids
Total Solids

26.1 26.2 29.0

Enzyme
Total Solids

28.6
Liquefaction conc. =1.5%
24.4 28.3
24.4 Time = 60 min
27.9
27.5
22.6 22.6
77.8 2.1

68.9 1.8
2.1 32.97 77.8 32.97
1.8 31.49 68.9 31.49 60.0 1.5
1.5 30.00 60.0 30.00
51.1 1.2
C: Liquefaction time51.1 28.51 A: Slurry Conc. C: Liquefaction time B: Enzyme Conc.
B: Enzyme Conc. 1.2 28.51 A: Slurry Conc.
0.9 27.03 42.2 27.03 42.2 0.9

(b)

Slurry Conc. =30%

Liquefaction Enzyme 9.4
8.5 8.5
Consistency index

Time = 60 min conc. =1.5%

Co n siste n cy in d e x
Consistency index

6.8 7.4
6.5

4.6 5.1 5.3

2.6 3.3 3.3

0.7 1.6 1.3

2.1 33.0 77.8 33.0 77.8 2.1

1.8 31.5 68.9 31.5 68.9 1.8

1.5 30.0 60.0 30.0 60.0 1.5

B: Enzyme Conc.1.2 28.5A: Slurry Conc. 51.1

C: Liquefaction time 28.5A: Slurry Conc. 51.1
C: Liquefaction time 1.2B: Enzyme Conc.

0.9 27.0 42.2 27.0 42.2 0.9

(c)
Fig. 1 Response surfaces showing effect of slurry concentration, enzyme concentration and liquefaction time on a yield, b total solids and c
consistency index of oat milk
614 Food Bioprocess Technol (2014) 7:610–618

added as a catalyst for the enzyme. These samples were regression coefficient for linear, quadratic and interaction
treated with different levels of enzyme concentration and terms, respectively, and ε is the error. Optimization of the
liquefaction time for optimization of the oat milk fitted polynomials was done using a desirability function
manufacturing process at 75 °C. The liquefied oat solids (Derringer and Suich 1980). The optimum condition was
were then filtered through muslin cloth to get the oat milk. verified by conducting experiments at that condition.
At the end of the treatment, the enzyme was inactivated by Responses were monitored and results were compared with
heating at 100 °C for 5 min. Total solids were estimated by model predictions.
AOAC method 990.20 using the principle of gravimetric The fitted polynomial equation was expressed as contour
analysis (AOAC 1990). The total yield was estimated as plots using the design expert software (version 8) in order to
the percentage of filtrate obtained. visualise the relation between the response and experimental
A Bohlin Gemini Rheometer (Malvern Instruments levels of each factor and to deduce the optimum condition
Limited, UK) was used to perform the rheological measure-
ments. Rheology of different samples was measured using
plate and cone geometry with a cone diameter of 20 mm and Results and Discussion
a cone angle of 2° in the shear rate range of 0.1 to 100 s−1.
The samples were adequately mixed using a magnetic stirrer The responses viz. oat milk yield, consistency index and total
to achieve uniform consistency before loading into the rhe- solids under different treatments are given in Table 2. The
ometer. The sample was brought into the lower plate using a functional relationship between the independent and depen-
spatula and filled up the gap by lowering the upper cone dent parameters was modelled using second order polyno-
down to the designated gap. mial equation. The coefficient of the actual functional rela-
tionship for predicting A, B and C are presented in Table 3.
Experimental Design and Statistical Analysis The insignificant terms were removed on the basis of
Student’s t ratio. The results were analysed using ANOVA
The central composite rotatable design (CCRD) was used in (Table 4). It is evident from the table that the first and second
designing this experiment and optimization was done through terms were found out to be significant and the lack of fit was
RSM. Three independent variables namely slurry concentration
(x1), enzyme concentration(x2) and liquefaction time (x3) were Table 5 Power law parameters for oat milk formulations
chosen. The ranges of parameters for the present investigation
Experiment no. Consistency indexa R2 n Chi square
were determined from preliminary experiments done. The ex- (k) ( Pa sn )
perimental design in the coded (x) and actual (X) levels of
variables is shown in Tables 1 and 2. All the experiments were 1 8.35±0.03 0.9346 0.3518 1.89E+05
performed in triplicates. The responses function (y) measured 2 1.40±0.02 0.9649 0.4516 3,095
were yield, total solids and consistency index of oat milk. The 3 1.74±0.03 0.9228 0.5832 2.16E+05
analysis of variance (ANOVA) tables were generated and the 4 5.12±0.01 0.9378 0.3273 3,158
effect and regression coefficients of individual linear, quadratic 5 2.98±0.04 0.918 0.4653 2.35E+05
and interaction terms were determined. The significances of all 6 6.78±0.03 0.9119 0.3759 1.84E+05
terms in the polynomial were judged statistically by computing 7 5.94±0.05 0.9213 0.3267 1.02E+04
the F value at a probability (p) of 0.001, 0.01 or 0.05. The 8 4.32±0.01 0.9324 0.3314 3,654
regression coefficients were then used to make statistical cal- 9 9.83±0.02 0.8476 0.2924 3.77E+05
culation to generate contour maps from the regression models. 10 5.34±0.03 0.8998 0.3213 2.23E+04
A second-order polynomial equation was used to fit the 11 8.42±0.03 0.8778 0.312 4.04E+05
experimental data given in Table 2. The model proposed for 12 4.99±0.04 0.9189 0.2969 1.20E+04
the response (Y1, Y2 and Y3) was: 13 4.74±0.01 0.9126 0.3254 2.02E+05
14 6.04±0.05 0.8868 0.3447 3.97E+05
X
i¼3 X
i¼3 X X 15 10.22±0.04 0.8564 0.293 3.71E+05
Y i ¼ β0 þ βi X i þ βii X 2i þ i< j
βij X i X j
i¼1 i¼1
16 7.19±0.04 0.8914 0.3519 2.45E+05
17 1.03±0.05 0.9125 0.3651 4.59E+03
þε ð1Þ 18 5.09±0.01 0.905 0.3789 2.63E+05
19 1.62±0.01 0.8354 0.4459 3.70E+05
where Yi (i=1 to 3) is the predicted response for slurry 20 5.66±0.02 0.8901 0.3562 2.86E+05
concentration, enzyme concentration and liquefaction time,
respectively; β0 is the value of the fitted response at the a
Mean±standard deviation
centre point of the design, and β i , β ii and β ij are the (n=3)
Food Bioprocess Technol (2014) 7:610–618 615

Fig. 2 Shear rate versus shear

stress for the 20 sample
formulation as per the
experimental design
presented in Table 2

insignificant. The lack of fit signifies the failure of the model
to represent data in the experimental domain. The model is a
good fit as represented by the high value of coefficient of
determination (R2).
Effect of Enzyme Concentration, Slurry Concentration
and Liquefaction Time on Yield of Oat Milk
The yield of oat milk can be increased using various pre-
treatments, i.e. particle size reduction, hydrothermal treat-
ment and enzymatic treatment, etc. Enzymatic treatment
alone or in combination with other is one of the potential
treatments which results in increased yield with better oat
milk quality and acceptability.
The oat milk yield is found to be a function of the linear
and quadratic effects of enzyme concentration. It can be seen
from Fig. 1a that an increase in enzyme concentration
resulted in increased yield of oat milk. This can be mainly
due to the action of enzyme on oat starch by the process of
liquefaction. The conversion of starch to dextrin results in
decrease in viscosity of oat milk thereby making filtration
process easier. Treatment time is also an important process
parameter because it may influence the liquefaction process
and oat milk yield, i.e., the longer the enzyme can react with
substrate, the greater the conversion of starch to dextrins, up
to a certain limit (Choi et al. 2010)
The oat milk yield varied from 53.92 to 78.87 % from
enzyme treated oat slurry. Thus the consistency index varied

Fig. 3 Superimposed contour

Design-Expert® Software
Overlay Plot
plots showing the shaded 2.5
overlapping area for which
Overlay Plot
yield≥75 %, total solids≥25 %
and consistency index value of Yield Consistency index: 1.03
1.03 Consistency index Yield: 75
Total Solids 2.0

X1 = A: Slurry Concentration
X2 = B: Enzyme Concentration

Actual Factor Total Solids: 25

1.5
C: Liquefaction time = 49.0

1.0

0.5
25.0 27.5 30.0 32.5 35.0
X1: A: Slurry Concentration
X2: B: Enzyme Concentration
616
Table 6 Predicted and actual responses for optimised process
Responses Predicted Experimental value
Total yield (wt%) 76.90 77.59±0.12
Total solids (wt%) 25.45 25.98±0.15
Consistency index (Pa sn) 1.03 1.01±0.08
a
Mean±standard deviation
n=3
from 10.22 to 1.03 Pa sn due to the action of enzymes on
starch. Increase in yield and decrease in viscosity are mainly
due to the action of liquefying enzyme on the starch present
in the oat flour (Singh et al. 2000; Lucyna et al. 2003).
Analysis of variance was carried out considering second
order polynomial equation (Eq. 1) on the responses to eval-
uate the adequacy of fit and results are reported in Table 4.
Calculated F values of yield were higher than its tabulated F
value at 1 % level of significance. The coefficient of deter-
mination (R2 values) for the developed model was 0.91.
Effect of Enzyme Concentration, Slurry Concentration
and Liquefaction Time on TSS of Oat Milk
The effect of slurry concentration and enzyme concentration
on the total solids (TSS) content in oat milk is shown in
Fig. 1b. The amount of total solids is important as it affects
the end quality of product formulated (Trinh et al. 2007). It is
clear from the graph that slurry concentration plays a major
role in the content of total solids present in oat milk. It can
also be seen from the ANOVA data (Table 4) that slurry
concentration is the most important factor playing role in
the total solids content (p<0.001). As the slurry concentra-
tion was increased, total solids concentration in the oat milk
was found to increase as expected. The positive linear and
quadratic effects of slurry concentration are significant at
p<0.001, which explains the observed nature of the curve
as shown in Fig. 1b. The other quadratic terms are not
significant. It is interesting to note that the interaction is not
significant. A higher slurry concentration would impose
additional handling, processing difficulties during produc-
tion process and increased cost of raw material. Also, under
Fig. 4 Optimised process
flowchart for the production of
oat milk
Rolled Oats (1Kg)
Food Bioprocess Technol (2014) 7:610–618
the range of independent variables taken for the experiment,
a an enzyme saturation level may reach at lower level of slurry
concentration after which increase in concentration of slurry
will not have any effect on enzymatic process but will only
lead to difficulty in filtration process. Hence, an optimised
condition of independent variables is very important to
achieve total solids in desirable ranges.
Effect of Enzyme Concentration, Slurry Concentration
and Liquefaction Time on Consistency Index of Oat Milk
The effect of slurry concentration and enzyme concentration
on the consistency index is shown in Fig. 1c. The power law
model was found to fit the experimental data with high
degree of correlation coefficient (R2 =0.89 to 0.96). The
power law model can be explained by the equation:
τ ¼ kγ n ð2Þ
Where, t is the shear stress, γ is the shear rate and k is the
consistency index (Pa sn) and n is the flow behaviour index.
The values of consistency index (k) and the flow behaviour
index (n) resulting from the fits are given in Table 5.
It is understood from the experimental data that with the
increase in the shear rate the apparent viscosity decreased for
all the samples investigated. The flow of the particle arising
from the particle–particle interaction can result in a decrease
in viscosity. It can be also related to the reduction in the size
of the colloidal aggregates with the increase in the shear rate
(İbanoğlu 2002). This can be attributed to colloidal nature of
oat milk. When shear force is applied, the particles may
rearrange themselves into parallel direction with shear force
and big particles may break into small particles by shear
force, thereby decreasing the viscosity (Charm 1963).
The power law parameters of all the formulations are
shown in Table 5. The consistency index is the indicator of
the viscous nature of the system (Simuang et al. 2004). For
all the samples in this experiment, consistency index (k)
decreased with the increase in enzyme concentration. This
signifies the reduction of the apparent viscosity (ɳ app) with
the increase in the enzyme concentration. All of the treat-
ments in the experiment presented behaviour typical of non-
α- Amylase
(77.78mg/Kg of Rolled oats)
Enzymatic
Hydrolysis
3.70Kg
Mixing Oat slurry Filtration Oat
Milk(2.85Kg)
Water (2.70Kg)
Filter Cake(0.86Kg)
Food Bioprocess Technol (2014) 7:610–618
Newtonian fluids, i.e. the relationship between shear rate and
shear stress was not linear. The flow behaviour index (n)
obtained from the experiment indicated that the oat milk after
enzyme treatment had a pseudo-plastic behaviour. Figure 2
shows the relationship between the calculated shear rate and
shear stress for all the different treatments. The magnitude of
the value of the flow behaviour index (n) was between
0.2924 and 0.5832. The value of n for different treatments
can be seen in Table 5. The statistical analysis showed no
significant difference (p<0.05) between flow behaviour in-
dex value at different enzyme concentration.
Optimization of Process Parameters
The optimum process conditions were determined by super-
imposing the contour plots of all the responses and finding
the region where operating conditions were ‘near’ optimal
for all the responses. Figure 3 shows the optimum conditions
of the process to give yield ≥ 75 %, total solids ≥ 25 % and
consistency index as 1.033 Pa sn, respectively. There are a
number of combinations of variables that could give maxi-
mum levels of yield, total solids in the desired range and
targeted value of consistency index. The desirability function
was used to combine multiple responses into one response
called the “desirability function” by choice of value from
0 (one or more product characteristics are unacceptable) to 1
(all product characteristics are on target)(Smith 2005). The
method is attractive because it is intuitive and simple. The inputs
are mean response estimates, target value, and upper and lower
acceptability bounds. The individual desirability is combined
using the geometric mean. The basic idea of the desirability
function approach is to transform a multiple response problem
into a single response problem by means of mathematical trans-
formations. The desirability function involves transformation of
each estimated response variable Yj to a desirability value dj,
where 0 ≤ di ≤ 1. The value of di increases as the “desirability”
of the corresponding response increases.
Optimization was performed using a multiple response
method called desirability. This optimization method incor-
porates desires and priorities for each of the variables devel-
oped a procedure for specifying the relationship between
predicted responses on a dependent variable and the desir-
ability of the responses.
Based on the results, it can be asserted that the oat milk
production was not dependent on a single independent factor
but interaction of the different factors was important in the
optimization of the oat milk production. As a result of the
optimization, the predicted values of the responses were:
27.0 % (w/w) slurry concentration, 2.1 % (w/w) enzyme
concentration and liquefaction time of 49.0 min. The calcu-
lated desirability for this formulation was 0.96.
The adequacy of the model equations for predicting optimum
response values was tested using slurry concentration 27.0 %,
617
enzyme concentration 2.1 % and liquefaction time of 49.0 min
(Table 6). This set of conditions was determined to be optimum
by the RSM optimization approach and was also used to validate
experimentally and predict the values of the responses using the
model equation. The results showed that the experimental values
were in agreement with the predicted value.
Conclusions
The oat milk was successfully produced, using response
surface methodology as an optimization tool with 1 kg of
rolled oats resulting in 2.85 kg of oat milk having total solids
in the range of 25.01±0.15 and consistency index value of
1.01±0.08 Pa sn. The optimised process flowchart for the
production of oat milk with mass balance is shown in Fig. 4.
The different conditions (slurry concentration, enzyme con-
centration and liquefaction) for enzyme treatment revealed
that all these variables markedly affect the yield, total solids
and rheological parameters of the oat milk. These can be
related to the enzyme treatment conditions by second order
polynomials. Using the contour plots, the optimum set of the
operating variables are obtained graphically in order to ob-
tain the desired levels of these properties of the oat milk. The
optimum conditions for making oat milk were 27.0 % (w/w)
slurry concentration, 2.1 % (w/w) enzyme concentration and
liquefaction time of 49.0 min.
Acknowledgment The authors would like to acknowledge the De-
partment of Biotechnology, Government of India, for financial support.
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