Agricultural and Biological Chemistry

ISSN: 0002-1369 (Print) (Online) Journal homepage: http://www.tandfonline.com/loi/tbbb19

Denaturation Temperature of Soy Protein under

Low Moisture Conditions

Naofumi Kitabatake, Mineo Tahara & Etsushiro Doi

To cite this article: Naofumi Kitabatake, Mineo Tahara & Etsushiro Doi (1989) Denaturation
Temperature of Soy Protein under Low Moisture Conditions, Agricultural and Biological Chemistry,
53:4, 1201-1202, DOI: 10.1080/00021369.1989.10869430

To link to this article: https://doi.org/10.1080/00021369.1989.10869430

Published online: 09 Sep 2014.

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Agric. Bioi. Chern., 53 (4), 1201 ~ 1202, 1989 1201

Short Communication superspeed centrifuge SCR 20B with an angle

rotor (RPR 20-2)] for 10 min at room tempera-
Denaturation Temperature of Soy ture, the pH of the supernatant obtained was
adjusted to 4.5 by the addition of 6 N HCI. The
Protein under Low Moisture suspension was left for 15 min at room tem-
Conditions perature, and then centrifuged again at
10,000 rpm. The precipitate was collected and
Naofumi KIT ABA TAKE, Mineo T AHARA suspended in distilled water. The pH of this
and Etsushiro Dm sample was adjusted at pH 7.5 with 6 N NaOH.
Research Institute for Food Science, Kyoto University,
Then the pH of the sample was adjusted to pH
Uji, Kyoto 611, Japan 4.5, and the sample was centrifuged and neu-
Received January 9, 1989 tralized as above. This procedure was repeated
twice more to remove the whey protein from
the isolate. The soy protein isolate obtained
The thermal denaturation of protein has (pH 7.5) was lyophilized.
been studied extensively from various points of The soy protein powders obtained were
view, but in most experiments a diluted protein dissolved in distilled water to prepare the
solution in water has been used as the sample. protein sample for analysis. Protein concen-
The heat-stability and denaturation of protein trations were assayed by the method of
under low moisture conditions are not well Bradford4 l using a Coomassie protein assay
understood. In food processing and manufac- reagent (Pierce Chemical Company, Rock-
turing, as in extrusion cooking, foodstuffs ford, IL). Bovine serum albumin was used
including protein are subjected to various heat as a standard.
treatments under low moisture conditions. The The thermal denaturation of soy proteins
denaturation temperature (TD) of protein in a under various conditions was studied by differ-
solution is usually below 100°C. The TD of ential scanning calorimetry (DSC) on a differ-
protein is generally independent of the protein ential scanning calorimeter (DSC 100) con-
concentration, but at extremely high concen- nected to aT A station (Seiko Instruments Inc.,
trations of protein (that is, with a low water Tokyo). The silver pans (Seiko Instruments,
content), TD increases 1 - 3 ) to about 120oC. No. 560-003) used had been heated in an
This increase in TD is not found in the range atmosphere saturated with water vapor at
above 30% (w/w) water, but below it. Here, we 170°C for 1 hr.
observed that the TD values of soy proteins The sample for the DSC of soy protein
increased linearly with the decrease in the under low moisture conditions was prepared
water content of the protein sample and as follows. First, 50 J1l of soy protein solution
reached about 170oC with 29% water. This (70 mg/ml) was put into the silver pan, and
means that soy protein, which is widely used as then the pan was put in the bottle for lyophili-
a food material for textured vegetable protein zation. A given volume of distilled water was
with extrusion cooking, needs a much higher added to each pan for a given water content
temperature than lOOoc to be denatured under and then the pan was sealed. The closed pan
low moisture conditions. was left for 24 hr at room temperature for
Soy protein isolate was prepared from de- tempering the protein with water. The pans
fatted soybean flour (Fuji Oil Co., Ltd., were placed in the DSC cell and heated to
Osaka, Japan) as follows. To the defatted 180°C or 200°C at a programmed rate of
soybean flour was added fifteen volumes of increase of 5°C/min. The sampling time of the
distilled water, and the mixture was stirred for data was 0.5 sec. The DSC instrument was
30 min at room temperature. After centrif- calibrated with indium. The data were stored
ugation at 15,000 rpm (28,000 x g) [Hitachi in a diskette and analysed with built-in soft-
 1202 N. KITABATAKE, M. TAHARA and E. DOl

170.TC and 135.6°C. The decrease in the

water content increased the denaturation tem-
perature of each protein. The inset in Fig. I
~ (a)

..
;:
Ill
G>
shows that the correlation between the water
content and the denaturation temperature was
.1: (b)
good. This relation has not been observed for
u other proteins; the T 0 values reported for
·e.. other proteins were below 120°C even when
G> the water content was below 30%. 1 - 3 ) In this
.1:

1 r~1~1
sense, soy proteins have different characteris-
tics from those of other proteins. The total
heat involved in the denaturation process
(L1H) could be calculated from the same ther-
mograms. The L1H values of soy proteins were
! 0
o 20 40 60 ao 100
not different under three moisture conditions.
Water content (% w/w)

To produce textured vegetable protein with

60 80 100 120 140
Temperature (°C)
Fig. 1. DSC Thermo grams of Soy Protein Isolate in the
Temperature Range 60 ~ 200oc.
Moisture content (% wjw) is (a) 94%, (b) 64%, and (c)
29%. In the inset, the denaturation temperature of each
peak (the top of each peak) is plotted against the water
content of the sample.
ware designed for DSC analysis including L1H
and T 0 calculations. The water content of the
sample was measured with a Karl-Fischer
moisture meter (Model MCI KF-05, Mitsu-
bishi Kasei Co., Tokyo).
Figure I shows the thermograms of soy
protein isolat~ at different water contents. Two
endothermic peaks below lOOoc were observed
at high water content (94% wjw) (a 93.3°C and
76.SOC). The peaks seem to correspond to the
peak of the 11 S globulin and that of 7S
globulin of soy protein. 5 l With a decreased
water content, these two peaks were shifted to
higher temperatures; the decrease in moisture
increased the denaturation temperature of
each protein. At 64% water, the two T 0 values
were above 100°C (at 124.9°C and 102.3"C).
160 180 200
extrusion cooking from raw plant protein
(usually soy protein), heating at around 150'~C
for 30 to 60 sec is known empirically to be
needed when moisture is 15 ~40%. 6 ) However,
the changes taking place at the molecular level,
the reason why such a temperature is needed,
and whether soy protein melts at these tem-
peratures have been unknown. The results of
this study suggested that the temperature need-
ed for the extrusion cooking of soy protein to
texturize was the denaturation temperature of
soy protein itself under low moisture con-
dition. Further details will be reported later.
References
I) M. Luescher, M. Ruegg and P. Schindler, Rio-
polymers, 13, 2489 (1974).
2) M. Ruegg, U. Moor and B. Blanc, Biochim. Biophys.
Acta, 400, 334 (1975).
3) Y. Fujita andY. Noda, Int. J. Peptide Protein Res.,
18, 12 (1981).
4) M. Bradford, Anal. Biochem., 72, 248 (1976).
5) S. Damodaran, J. Agric. Food Chern., 36, 262 (1988).
6) D. A. Ledward and J. R. Mitchell, "Food Structure:
Its Creation and Evaluation," ed. by J. M. Blanshard
and J. R. Mitchell, Butterworths, London, 1988, pp.
219~229.

At 29% water, the two peaks were found at