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Food Flavors: Formation, Analysis and Packaging Influences
© 1998 Elsevier Science B.V. All rights reserved 621
Abstract
Soybean trypsin inhibitors (TI) in soymilk were treated by heat and ultrasound of 20 kHz.
The influence of several factors (temperature, time of treatment, pH, ultrasonic power,
soymilk concentration, and ionic strength) on inactivation of TI was investigated. The results
of the experiment shown that temperature was the major factor to affect the inactivation of TI,
and treatment time was the next important factor. Under the experimental conditions of
temperature 80 °C, ultrasonic power of 150 w, and pH 7.0 for 5 min treatment, TI in soymilk
sample could be inactivated by 73%. The retained 27% TI was difficult to inactivate. This
residual component is mainly Bowman-Birk inhibitor (BBI) which is extremely stable to heat
as well as ultrasound of 20 kHz.
1. INTRODUCTION
ultrasonic wave amplitudes, and the static pressure did not seem to significantly affect the
destructive effect of the process. Lopez and Burgos [10] further investigated the effects of
sonication physical parameters, pH, KCl, sugers, glycerol and enzyme concentration on the
inactivation of soybean lipoxygenase (LOX), an enzyme involved in off-flavor development
in vegetable products, under MTS operation. Their resuhs suggest that MTS inactivates LOX
by two different mechanisms, one associated with heat and the other with ultrasound. They
also found that the effect of static pressure on the inactivation of LOX is not substantial.
Our objective is to investigate the feasibility of applying ultrasound in combination with a
mild heat treatment on the inactivation of TI in soymilk at atmospheric pressure and the
effects of various influent factors such as temperature, duration of ultrasound treatment,
ultrasound power level, pH value of the solution, soy solid concentration and ionic strength.
pulse with the on-off ratio of 3:2 second. At the end of the treatment, the cylinder with
soymilk was immediately immersed into ice water. The treated sample was then diluted and
the residual TIA was assayed.
2.5. Assay for trypsin inhibitor activity (TIA)
A modification of the Kakade's procedure, developed by Smith et al. [11], using the
synthetic substrate benzoyl-DL-arginine-p-nitroanilide (BAPNA), was employed to measure
TIA. The method involves extraction of the inhibitors from the sample at pH 9.5 and mixing
unfiltered suspensions with bovine trypsin. The activity of the remaining trypsin is then
measured by offering it BAPNA under standard conditions. The p-nitroaniline released is
measured spectrophotometrically at 410 nm. This provides a linear measure of the residual
trypsin activity, so that the amount of pure trypsin inhibited per unit weight of sample can be
calculated.
such as heat, acids and alkaUs. The experimental results indicated that BBI is also extremely
stable towards the effect of ultrasound. Therefore, under the operation of ultrasound, about
30% of residual TIA is always difficult to inactivate.
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4. CONCLUSIONS
This study shows that the appHcation of ultrasound with mild heat treatment at atmospheric
pressure can facilitate the inactivation of TI. Treatment temperature is the most important factor
in such operation. Other influent factors are sonification time, ultrasound power, pH of the
solution and ionic strength. Under the condition of temperature 80 °C, ultrasonic power 150 W,
and pH 7.0, about 73% of TI was inactivated for 5 min treatment. About 27% residual of TI was
difficult to inactivate. This residual component is mainly Bowman-Birk inhibitor (BBI) which is
extremely stable to the ultrasound of 20 kHz.
5. ACKNOWLEDMENT
This work was financially supported by the Research Committee of Hong Kong Polytechnic
University.
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