Trypsin Inhibitor and Urease Activity of Soybean Meal Products from Different Countries and Impact of Trypsin Inhibitor on Ileal Amino Acid Digestibility in Pig

Trypsin inhibitor (TI) and urease activity (UA) are the most relevant quality aspects of soybean meal (SBM) for monogastric animals. This study’s objective was to determine the levels of TI and UA in SBM collected from feed mills in different countries, whether TI and UA are correlated since they are both heat-labile, and impact of TI on pig ileal amino acid digestibility. TI was 1.20–8.37 mg g⁻¹ with 36.4% samples <3 mg g⁻¹, UA was 0–0.311 ΔpH unit with 64.2% samples <0.05 ΔpH unit in 847 solvent-extracted SBM samples. TI and UA results varied by countries and world areas. By country, mean TI was highest in Germany and lowest in Philippines, mean UA was highest in India and lowest in Philippines. By world areas, mean TI was highest in Latin America and lowest in North America, UA was highest in Asia and lowest in Latin America. TI and UA results vary in different soybean products, TI ranked as raw soybeans > expeller SBM > full-fat-extruded (FFE) SBM ≥ heat-inactivated full-fat (HIFF) SBM > solvent-extracted SBM; UA ranked as raw soybeans > HIFF SBM > FFE SBM ≥ solvent-extracted SBM ≥ expeller SBM. TI was not correlated with UA in 847 solvent-extracted SBM, suggesting that UA may not be used as a surrogate indicator for TI. A diet contained 38% SBM with 8.78 mg g⁻¹ TI significantly reduced apparent ileal digestibility of amino acids and crude protein by 13.3–26.0% and 23.3% points, respectively, in cannulated pigs compared to SBM with 2.51 mg g⁻¹ TI.     

Trypsin Inhibitor Assay: Expressing,Calculating, and Standardizing Inhibitor Activity in Absolute Amounts of Trypsin Inhibited or Trypsin Inhibitors

For expressing trypsin inhibitor activity (TIA), trypsin units inhibited (TUI), trypsin inhibited, and trypsin inhibitors have been used. Although the last two units are preferred, their calculations in current practices require refinement. With the proposed AOCS method Ba 12a-2020, four experiments were conducted, using four trypsin preparations having specific activity of 11,625, 12,602, 13,728, and 14,926 Nα-benzoyl-L-arginine ethyl ester (BAEE) units/mg protein, respectively. Experiment 1 determined the relationship between absorbance at 410 nm (A410) and trypsin concentration. Experiment 2 involved assaying raw and heated soybeans, expressing TIA as TUI/mg sample and μg trypsin inhibited/mg sample, and determining conversion factors between the two units. Experiment 3 resembled Experiment 2 except for using purified soybean Kunitz inhibitor (KTI) and Bowman-Birk inhibitor (BBI). Conversion factors determined correlated highly with trypsin-specific activity (R² = 0.9789). After standardizing against a reference trypsin having 15,000 BAEE units/mg protein, a standardized conversion factor of 0.03 A410 (1.5 TUI) = 1 μg trypsin inhibited was determined. It remained consistent regardless of trypsin specific activity, with or without inhibitors, and type of inhibitor samples. By using purified inhibitors (Experiment 3), conversion values between TUI and μg trypsin inhibitor and between μg trypsin inhibited and μg trypsin inhibitor could also be calculated, enabling expression of TIA in amounts of pure KTI, BBI or their equivalents. Furthermore, when the AOCS method was modified with half substrate concentration, half trypsin concentration or half both (Experiment 4), TIA values in TUI could change with modifications but values in mg trypsin inhibited (standardized) or trypsin inhibitor remained consistent.     

Comparison of ISO 14902:2001 with AOCS Ba 12a-2020 for determining trypsin inhibitor activity in protein products

For measuring trypsin inhibitor activity (TIA), there are two major official methods: American Oil Chemists Society (AOCS) method Ba 12a-2020 and International Organization for Standardization (ISO) 14902:2001. The former was recently approved. The two methods differ in sample preparation, extraction, colorimetric assay systems and TIA calculations. In this study, the two methods were symmetrically compared using three unique sets of samples: assorted protein products of soybeans, pulses, and grains; soybeans boiled for varied durations; and soy white flakes toasted for varied durations. For given samples, significant differences existed in TIA measured by the two methods, resulting from effects related to the assay systems and TIA calculations, not from the difference in sample preparation and extraction. When the same trypsin was used, TIA (in mg trypsin inhibited/g sample) measured by the two methods were highly correlated (r = 0.9973, n = 27), giving an equation of y = 0.5464x − 0.4887, where y represents ISO values and x for AOCS values. The line connecting ratios of ISO/AOCS in TIA and AOCS values remained relatively flat around 0.53 but started to curve down when TIA approached the lowest. Furthermore, for the same samples, TIA values measured by the ISO method decreased with increasing specific activity of trypsin used, while AOCS values remained consistent, leading to decreasing ratios of ISO/AOCS. Therefore, accurate and direct comparison of the two methods was impossible. It could not be resolved by simply changing ISO method's calculations as hypothesized earlier. Regardless, for most samples, ISO values were roughly about 55% of AOCS values.     

Steaming,boiling after pre-frying,and stir-frying influence the fatty acid profiles and oxidative stability of soybean oil blended with docosahexaenoic acid algal oil

The objective of this study was to improve the content of docosahexaenoic acid (DHA) and obtain the blended oils used for different cooking methods (steaming, boiling, and stir-frying) by blending 0%–15% DHA algal oil into soybean oil. It was shown that the addition of DHA algal oil increased saturated fatty acid (SFA) (1.57%) but decreased monounsaturated fatty acid (MUFA) (0.76%) and polyunsaturated fatty acid (PUFA) (0.68%). Various cooking methods significantly changed the fatty acid (FA) compositions. Steaming is a more effective way to prevent DHA loss and the production of trans-fatty acid than boiling and stir-frying. Besides, a positive result from free fatty acid (FFA) and peroxide value also demonstrated that steaming was a better way to protect oils. Overall, the soybean oil blended with 3% DHA algal oil with better oxidative stability and could be recommended for daily application by steaming.     

The comparative value of heated ground unextracted soybeans and heated dehulled soybean flakes as a source of soybean oil and energy for the chick

Experiments were conducted to evaluate heated unextracted soybean fractions as sources of soybean oil and protein for the growing chick. Heated dehulled unextracted soybean flakes produced growth rate and feed efficiency equal to that obtained with the combination of soybean oil meal and degummed soybean oil while heated ground unextracted soybeans were less satisfactory in this respect. The poorer results obtained with ground unextracted soybeans were shown to be related to a poorer absorbability of the oil in them. Flaking the soybeans markedly improved the absorbability of the oil by the chick, probably by causing a greater disruption of cellular structure than was obtained by the grinding of the soybeans. The metabolizable energy of ground unextracted soybeans was substantially less than that of unextracted soybean flakes. Most of the differences in metabolizable energy were accounted for by differences in absorbability of the oil. Soybean hulls at a level equivalent to that contained in soybeans were found to have no effect on growth rate and only a slight effect on feed efficiency. Autoclaving soybean oil did not lower its value for the chick. The relationship between the poorer growth obtained with ground unextracted soybeans and the low absorbability of the oil in them was discussed. To obtain maximum efficiency in the use of unextracted soybean products in chick rations, some such means as flaking must first be employed to increase the availability of the oil.     

Lipid oxidation products of heated soybeans as a possible cause of protection from ruminal biohydrogenation

Heating oilseeds has been shown to improve the milk fatty acid profile when given to dairy cows, compared to raw oilseeds. However, results from published studies are conflicting. The conditions of heating and storage of the oilseeds could be responsible for these differences, probably partly through their effects on lipid oxidation, the products of which could act on ruminal biohydrogenation (BH). Thus, 15 different treatments were applied to ground soybeans: three levels of heating (no heating, 30 min at 110 or 150°C) × 5 ambient storage durations (0, 1, 2, 4, or 6 months). Soybeans were incubated in vitro with ruminal fluid for 6 h. Triacylglycerol (TAG) polymers, hydroperoxides and hydroxyacids (HOA), aldehydes, and fatty acids were assayed in soybeans and ruminal culture. No TAG polymer was detected in any treatment. Soybeans stored for a long time had a high content of HOA, whereas those heated at 150°C, whatever the storage duration, had high aldehyde contents. The percentage disappearance of cis‐9,cis‐12 18:2 and cis‐9,cis‐12,cis‐15 18:3 in incubates decreased significantly in cultures with heated soybeans, especially at 150°C, suggesting that this partial protection of polyunsaturated fatty acids (PUFA) from BH was at least in part linked to the aldehyde content of the heated soybeans. Practical applications: Oilseeds given to ruminants are often heated, and heat treatment is known to generate oxidation products. Knowing what oxidation products influence ruminal biohydrogenation of unsaturated fatty acids could result in technological processes allowing a better transfer of unsaturated fatty acids from oilseeds to ruminant products.     

Effect of soybean pretreatment on the color quality of soybean oil

Color reversion in soybean oil can be prevented by reducting the enzyme activity of soybeans before cracking and flaking. Soybean oil extracted from steamed, intact soybeans (18% moisture) had lower Rm (max. red) values in RBD oil, higher amounts of γ-tocopherol, plus its isomers, in both crude and RBD oil, and also higher amounts of hydratable phosphatides in crude oil than those in the oils from the same beans without steam treatment. For soybean pretreatments, a toasting process is less effective than the steaming process for the inhibition of color reversion of soybean oil. To prevent the occurrence of color reversion in RBD soybean oil, the amount of γ-tocopherol and γ-TED (5-[tocopheryloxy]-γ-tocopherol) should be above 550 ppm in crude oil.     

Effects of antioxidants,methyl silicone and hydrogenation on room odor of soybean cooking oils

Room odor characteristics produced by heated soybean oil (SBO) and soybean oils hydrogenated with copper (CuHSBO) and nickel (NiHSBO) catalysts were evaluated by a trained panel. Oils were intermittently heated to 190 C for total heating periods of 5, 15 and 30 hr. Oil additives investigated included methyl silicone (MS), tertiary butylhydroquinone (TBHQ) and a polymeric antioxidant in various combinations with citric acid (CA). In room odor tests directly comparing SBO, CuHSBO and NiHSBO, panelists rated the hydrogenated oils as having significantly less odor intensity than the SBO. The combination of CA+MS had the greatest effect in lowering odor intensity of the heated oils, followed by the mixture of CA+MS+TBHQ. The low odor intensity of the MS-treated oils remained fairly constant throughout the tests, while the higher intensity associated with all the other additive-treated oils decreased with increasing heating times, possibly as the result of formation of more volatile decomposition products in the initial heating stages. Methyl silicone had the strongest effect of any additive in decreasing objectionable room odors in the oils. Partially hydrogenated SBO treated with up to 5 ppm of MS produced cooking oils with low room odor intensity and low color development during prolonged heating.     

Changes in dielectric constant as a measure of frying oil deterioration

A soybean oil, a hydrogenated vegetable frying shortening and an animal-vegetable shortening were heated at 190 C for 8 hours each day for 4 days with and without the frying of potatoes. Samples were taken periodically and analyzed for various changes normally used to measure frying oil deterioration. The changes in the dielectric constant were determined with a patented instrument called the Food Oil Sensor. This instrument is standardized with 3. sample of the fresh oil, and it then measures the change in the electric capacitance of the heated oil samples. The dielectric constant of all three shortenings increased linearly with heating time. The greatest change occurred in the soybean oil sample and the smallest change in the hydrogenated vegetable shortening. For each shortening the increase was somewhat greater during frying than during heating without frying. Statistically significant correlations were obtained between instrument readings and increase in the total polar materials, the color, the peroxide values, the diene content, and the free fatty acids and the decrease in the iodine values.     

Absorption in rats of rapeseed, soybean, and sunflower oils before and following moderate heating

Rapeseed, soybean, and sunflower oil were heated for 15 min in a 5-mm oil layer in a pan at 180°C. The fatty acid composition was almost unaffected by heating, while the polymer content rose slightly and the tocopherol content decreased, except in soybean oil. The absorption of oils before and after heating was investigated in lymph-cannulated rats. Oils were administered as emulsions through a gastrostomy tube and lymph was collected during the next 24 h. The highest accumulated lymphatic transport of total fatty acids was observed after administration of rapeseed oil, and the lowest after heated sunflower oil. The accumulated transport was similar for all unheated oils. The transport of fatty acids was significantly lower in rats receiving heated oil compared to those receiving the corresponding unheated oil. Small increases in polymers may have contributed to the decreased lymphatic transport of oil following heating, although this probably does not fully explain the effect. The absorption of sunflower oil was more affected by heating than the absorption of soybean or rapeseed oil. Furthermore, the largest decrease in total activity of tocopherols following heating was observed in sunflower oil. Overall, these results demonstrate that the absorption of vegetable oils is affected by moderate heating.     

Dry extrusion as an aid to mechanical expelling of oil from soybeans

A new concept is described for mechanical extraction of oil from soybeans, using dry extrusion as a pretreatment. It was found that coarsely ground whole soybeans at 10 to 14% moisture could be extrusion cooked so that the extrudate emerges from the die in a semi-fluid state. The dwell time within the extruder was less than 30 seconds, and the temperature was raised to about 135 C. The semi-fluid extrudate was immediately pressed in a continuous screw press to obtain high quality oil and press cake. Extrusion prior to expelling greatly increased the throughput of the expeller over the rated capacity. An oil recovery of 70% was obtained in single pass expelling using pilot model expellers. Higher recovery rates can be expected with commercial scale expellers. The high temperature-short time extrusion cooking process eliminates the prolonged heating and holding of raw material in conventional expelling. Under the experimental conditions, press cake with 50% protein, 6% residual oil and 90% inactivation of trypsin inhibitors was obtained. The low fat cake was easily ground in a hammer mill without the usual problems associated with milling of whole beans. The expelled oil was remarkably stable with an AOM stability of 15 hr, which is comparable to refined deodorized oil according to NSPA specifications. The new procedure offers potential for producing natural soybean oil and food grade low fat soy flour by a relatively low cost operation. It may be adopted as an improvement to existing conventional expelling operations in less developed countries or as a commercial or on-farm operation for producing value added products from soybeans within the U.S.     

Factors affecting the content of tocopherol in soybean oil

The state of soybeans prior to extraction affected the tocopherol content of crude soybean oils. Soybean flakes with a thickness of 0.16–0.33 mm had higher extracted oil yield but a slightly lower tocopherol content of the oils than did cracked beans and thicker bean flakes. Highmoisture content and long storage of soybeans resulted in lower tocopherol content in the crude oils, with moisture content being more important than storage time at decreasing the tocopherol content of oils. Soybean oil from stored beans with 15±1% moisture content led to a more significant decrease in the tocopherol content than did oil from stored beans with low (12%) or high (18%) moisture contents. Soybean flakes contaminated with oxidized oil had a significant effect on the decrease of the tocopherol content in crude oils. The high amount of phospholipids in crude soybean oil might result in a smaller decrease in the tocopherol content of oil during heating.     

Microwave Super‐Heated Boiling of Organic Liquids: Origin,Effect and Application

This paper reports the state of the art of the microwave super‐heated boiling phenomenon. When a liquid is heated by microwaves, the temperature increases rapidly to reach a steady temperature while refluxing. It happens that this steady state temperature can be up to 40 K higher than the boiling point of the liquid. With the same reactor, overheating is not observed under conventional heating. The bulk temperature of a microwaved solvent under boiling depends on many factors: physical properties of the solvent, reactor geometry, mass flow, heat flow, and electric field distribution. The influence of these factors is studied and discussed. The kinetics of homogeneous organic reactions shows an extension of Arrhenius behaviour into the super‐heated temperature region. Reaction rate enhancement of order 10–100 can thus be achieved, which is normally only possible under pressure. Finally, we present a model predicting reaction kinetics and yields under classical and microwave heating, based on predicted temperature profiles in agreement with experimental data.     

Effect of Heating Time on the Thermal Stress Resistance of a Soda-Lime-Silica Glass

A model study was conducted of the thermal fatigue and single-cycle thermal shock resistance of indented soda-lime-glass rods heated over durations sufficiently short that thermal equilibrium was not reached prior to quenching into a water bath. The experimental results indicate that for short heating periods thermal fatigue life at fixed ΔT and the critical quenching temperature difference (Δ7_c) for a single-cycle quench significantly exceed the corresponding values obtained for specimens heated for sufficiently long periods that conditions of final thermal equilibrium were obtained prior to quenching. The implications of these results for single-cycle and multicycle thermal shock testing are discussed.     

非均匀受热条件下螺旋管内流动沸腾换热特性

目前对螺旋管在其管外表面均匀受热,管内两相流动换热的研究已十分丰富;但是在其管外表面非均匀受热条件下,管内两相流动沸腾换热特性的研究鲜有报道。为了解决螺旋管在实际运用中遇到的非均匀受热问题、得到其换热特性,本文采用了实验的方法研究了卧式螺旋管周向非均匀受热条件下管内流动沸腾换热特性。其中实验工况范围为系统压力P=0.7~1.0MPa,质量流速G=181~364kg/(m²·s),质量干度χ=0.07~0.69。实验考察了螺旋管管外壁在两种非均匀受热条件下管内的两相流动沸腾换热系数与热流密度、质量流速、质量干度的关系,并与管周向均匀受热工况进行了比较。结果表明,在螺旋管外壁面“外半周绝热、内半周受热”情况下管内流动沸腾换热系数值最大,而管外壁面“内半周绝热、外半周受热”情况下最小。     

环形通道内低Pe数高温液钠传热及其初始沸腾壁面过热度研究

对环形通道内低Pe数(20～70)的高温(300～700℃)液钠在由非旺盛湍流转变成旺盛湍流区内(Re=4000～17500)的传热进行了实验研究,并对正压下液钠初始沸腾壁面过热度进行了理论分析与实验研究.结果表明,所提供的计算关系式与实验数据吻合良好.实验采用的环管内径为6mm,外径为10mm,通道外壁绝热,内壁用高热流密度电加热元件加热.     

Lipoxidase deactivation to improve stability,odor and flavor of full-fat soy flours

Oxidation of soybean lipids catalyzed by lipoxidase was prevented by heat treatment of soybean meats, which were then ground to give a full-fat soy flour free of rancid odor and flavors. Our previous studies showed that lipids in cracked, dehulled, soybeans rapidly oxidized after the lipoxidase system was activated by increasing moisture content to 20%. A series of experiments are reported in which various heat treatments were evaluated for effectiveness of lipoxidase deactivation. Dry heat to 212 F, steaming, or both, deactivated lipoxidase to give flours that had low values of peroxide, conjugated diene and free fatty acid and had good flavors after 2 years’ storage. Wet heat alone was also an effective treatment, whereas wet heat preceded by dry heat at 180 F gave poor flavor stability after 2 years. Gas liquid chromatography studies gave evidence that the rapid formation of volatiles in full-fat soy flours was catalyzed by an enzyme system. A 10 member taste panel was able to detect significant flavor and odor differences between oxidized and nonoxidized samples. Presented at the AOCS Meeting, Chicago, October 1967. No. Utiliz. Res. Dev. Div., ARS, USDA.     

Minimizing protein insolubilization during thermal inactivation of lipoxygenase in soybean cotyledons

The objective was to develop a procedure for inactivating lipoxy-genase in soybean cotyledons without losing protein solubility. The approach was to moisten cotyledons to one of 2 levels in soft water or carbonate buffers, steam for a short time, hold for a definite period at a known temperature, cool and analyze for enzyme activity and protein solubility. Temperature dependence of both inactivation and insolubilization kinetics was determined. Increasing temperature of steaming and holding favored our objective. At 16.3% moisture, the pH 9.8 buffer was beneficial but the pH 10.8 buffer was not. The holding period was not beneficial compared to steaming alone. Recommended conditions were adjustment of cotyledon moisture to 16.3% with pH 9.8 buffer and then heating in steam for about 10 sec; at temperatures of 91 C and above, 99% of the lipoxygenase could be inactivated with retention of over 70% protein solubility. The effect of buffers on kinetics of heat inactivation and insolubilization appeared to be related to the states of hydration water, i.e., the presence of solute water at the higher moisture content.     

Functionality of dialyzed soybean extract

The soybean extract that was dialyzed against distilled and deionized water and adjusted to pH 7.5 (dialyzed soybean extract; d-SE) gave a transparent liquid after heating. The heated d-SE had less beany flavor and formed a translucent gel by heating again with 0.2 M NaCl. Gelation occurred at a lower protein concentration with NaCl than that of the original soybean extract. The dynamic viscoelasticity showed that the translucent gel from the heated d-SE was melted and gelled by the repetition of heating and cooling, indicating reversible gelsol transition. Cryoprecipitation could be avoided by the heating of d-SE. A gel was formed at 30°C by incubation of d-SE with NaCl. These properties of soybean extract may offer new applications as a food ingredient.     

High-temperature stabilities of oils from soybeans that lack lipoxygenases

Oils from normal or low-linolenic acid (18:3) soybeans that lack lipoxygenase (LOX) 2 or LOX 2 plus LOX 3 activities were evaluated for their stability during frying and for oxidative stability in bread cubes stored after frying. Soybean oils were extracted by a pilot-plant system and were refined, bleached, and deodorized in the laboratory. Citric acid was added to oils during the cool-down stage of deodorization. Two replications, separated at the point of conditioning, were evaluated for each genotype. Each sample (250 g) was heated to 180±5°C in a minifryer. Bread cubes were fried at the beginning of heating and after 20 h of heating. Heating of the oils was continued for 10 h each day for three consecutive days. Soybean oils with low 18:3 contents were significantly (P ≤ 0.05) more stable, as measured by conjugated dienoic acids and polymer values, than were oils with normal 18:3 contents. Low-LOX 2 or low-LOX 2 + 3 activity had no effect on peroxide values of soybean oils extracted from bread cubes. Sensory evaluation did not differentiate between oils that contained low or high 18:3 amounts or among oils from beans that lacked different LOX enzymes.