The biochemical and functional food properties of the bowman-birk inhibitor

The Bowman-Birk inhibitor (BBI) is a small water-soluble protein present in soybean and almost all monocotyledonous and dicotyledonous seeds. The molecular size of BBI ranges from 1,513 Da to about 20,000 Da. BBI is to seeds what alpha(1)-antitrypsin is to humans. Soy-based food products rich in BBI include soybean grits, soymilk, oilcake, soybean isolate, and soybean protein concentrate. BBI is stable within the pH range encountered in most foods, can withstand boiling water temperature for 10 min, resistant to the pH range and proteolytic enzymes of the gastrointestinal tract, bioavailable, and not allergenic. BBI reduces the proteolytic activities of trypsin, chymotrypsin, elastase, cathepsin G, and chymase, serine protease-dependent matrix metalloproteinases, urokinase protein activator, mitogen activated protein kinase, and PI3 kinase, and upregulates connexin 43 (Cx43) expression. Several studies have demonstrated the efficacy of BBI against tumor cells in vitro, animal models, and human phase IIa clinical trials. FDA considers BBI as a drug. FDA also approves labels claiming that consumption of at least 3 to 4 oz of tofu or 8 oz of soymilk or soy protein may reduce the risk of coronary heart disease and breast cancer. This review highlights the biochemical and functional food properties of the Bowman-Birk inhibitor.     

生物法失活大豆胰蛋白酶抑制剂的研究

对生物法失活大豆腋蛋白酶抑制剂进行了研究。测定了豆制品中胰蛋白酶抑制剂的活性;比较了外源蛋白酶失活胰蛋白酶抑制剂的能力,确定了碱性蛋白酶失活胰蛋白酶抑制剂的最优条件为pH值8.88～9.05、温度43.40～44.70℃、酶用量10.44～11.29μL/g、底物浓度6.51%～7.18%。以发芽12h的大豆加工的熟豆乳中胰蛋白酶抑制剂活性降低了83.2%。保加利亚乳杆菌(Lb)、米黑毛霉(M.M)和米根霉(R.O)发酵能有效失活豆乳中的胰蛋白酶抑制剂活性。     

胡萝卜、油菜的通电加热烫漂研究

研究了通电加热烫漂对胡萝卜、油菜中维生素含量的影响,并与普通电炉加热进行了比较;研究了胡萝卜、油菜通电加热烫漂的最佳工艺条件。实验证明,随着烫漂时间的延长,胡萝卜中的β-胡萝卜素含量明显降低,油菜中VC的含量也明显降低;通电加热烫漂后的维生素含量明显高于电炉加热;而胡萝卜通电加热的最佳烫漂工艺为,烫漂时间3 min,颗粒尺寸10 mm,料液比(g:mL)1∶4,电压220 V;油菜通电加热烫漂的适宜工艺条件为料液比(g:mL)1∶4,烫漂时间0.5 min,电压220 V。     

欧姆加热对豆浆风味物质的影响

为研究欧姆加热对豆浆挥发性物质的影响,采用顶空固相微萃取结合气相色谱-质谱联用技术,对豆浆的风味物质进行分析,利用内标法对各风味物质成分定量分析。通过对比50/30?μm二乙基苯/碳分子筛/聚二甲基硅氧烷（divinylbenzene/carboxen/polydimethylsiloxane,DVB/CAR/PDMS）、100?μm PDMS、65?μm PDMS/DVB 3 种萃取头对豆浆风味物质的萃取效果,筛选出最佳的萃取头,并在最适条件下分析豆浆的风味物质成分,为排除热效应对实验结果的影响,2?种热处理历程尽可能保持一致。结果表明：65?μm PDMS/DVB纤维萃取头较100?μm PDMS和50/30?μm DVB/CAR/PDMS纤维萃取头更适宜豆浆风味物质的测定;从豆浆样品中共检测到35?种挥发性物质,其中醛类物质16?种,醇类物质5?种,酮类物质3?种,酯类物质1?种,杂环烃类物质10?种,主要包括己醛、（E）-2-庚烯醛、1-辛烯-3-醇等,这些物质共同组成豆浆的特征香气;不同加热处理对豆浆风味物质种类的影响不大,但是对特征香气含量有显著影响（P＜0.05）;欧姆加热处理组样品中己醛、1-辛烯-3-醇、（E）-2-辛烯醛的含量分别比传统加热处理组约低45.55%、58.60%、25.56%,且在欧姆加热处理样品组中未检测到（E）-2-己烯醛和正己醇,而这些物质是豆腥味的主要组成成分。因此,利用欧姆加热方式加热豆浆可以显著降低豆浆的豆腥味。本研究可以为欧姆加热技术在豆浆加工业中的应用提供理论依据。     

Heat Inactivation of Trypsin Inhibitors in Soymilk at Ultra-High Temperatures

Soymilk samples at pH 7.5, 6.5 and 2 were subjected to heat treatment at 93°C and indirect ultra-high temperature. When heated at 93°C, 121°C and 132°C, trypsin inhibitor activity (TIA) in soymilk was more heat-labile at high pH than at lower pH. However, the effect of pH on rate of thermal inactivation was less pronounced when the holding temperature was increased to 143°C and 154°C. The point on a curve relating holding temperature and holding time, indicating inactivation of 90% of the TIA in soymilk at pH 6.5 in the range 93–154°C, coincided with the thermal-death-time curve of the organism putrefactive anacrobe 3679 at about 125°C.     

商品大豆饮料胰蛋白酶抑制剂活性的研究

本研究详细测定了未经热处理的生豆奶、各种经现代技术加工的大豆饮料产品、东方传统大豆食品豆腐等样品的胰蛋白酶抑制剂活性（ＴＩＡ）。其中生豆奶的ＴＩＡ（以每克蛋白质所抑制的纯胰蛋白酶毫克数表示）是６６．４ｍｇ／ｇ蛋白质，经巴氏杀菌的商品大豆饮料ＴＩＡ是２３．７ｍｇ／ｇ蛋白质。检验了经超高温灭菌处理的７种不同商标大豆饮品，结果表明这些饮品的ＴＩＡ在１３．３～３１．６ｍｇ／ｇ蛋白质之间。另有两种已知是经灭菌的大豆饮料，其ＴＩＡ最低，分别为４．１ｍｇ／ｇ蛋白质和７．７ｍｇ／ｇ蛋白质。又分析了一种采用东方传统加工方法生产的豆腐，其ＴＩＡ仅为６．４ｍｇ／ｇ蛋白质。上述这些分析结果反映了某些经现代加工方法生产的商品大豆饮料，在消除抗营养因子的处理上是不够充分的。     

豆浆中抗营养因子的去除方法

采用碱液浸泡、微波处理、加热处理、去皮等方法,对去除豆浆中抗营养因子的效果进行比较。其中,88℃恒温热处理10min可将豆浆中脂肪氧化酶活性降至常温下的14.20%,100℃恒温热处理20min可将豆浆中的胰蛋白酶抑制剂降低到9.96%。微波处理(2450MHz,800W)2min可以将脂肪氧化酶活性降至8.86%,处理8min可以将胰蛋白酶抑制剂活性降低到11.24%,处理4min即可将尿素酶活性降低至1%。相比于常规的热处理,微波处理可以更快速地破坏豆浆中的抗营养因子,缩短处理时间。此外,采用0.5%NaHCO3溶液浸泡、去皮等措施也可去除豆浆中的抗营养因子,可结合常规热处理、微波处理应用,以缩短加工时间。     

Immunoassays for Bowman-Birk and Kunitz Soybean Trypsin Inhibitors in Infant Formula

ABSTRACT: Because the consumption of soybean inhibitors of digestive enzymes in processed foods may have both beneficial and adverse health-related effects, reliable and rapid analytical methods for these inhibitors are needed. Monoclonal antibody-based sandwich enzyme-linked immunosorbent assays (ELISAs) were developed for the 2 major soybean protease inhibitors, the Kunitz trypsin inhibitor (KTI) and Bowman-Birk inhibitor (BBI) of trypsin and chymotrypsin. The ELISAs had limits of quantification of approximately 1 and 3 ng/mL for BBI and KTI, respectively, and were used to measure active inhibitors in soy infant formulas. Results were compared with enzymatic analyses and demonstrated that most of the trypsin- and chymotrypsin-inhibitory activities of infant formula were due to constituents other than KTI and BBI. The sandwich ELISA for BBI was also effective in detecting soybean germplasm with atypically low levels of BBI.     

甘薯块根中胰蛋白酶抑制剂研究进展

胰蛋白酶抑制剂(TI)是甘薯块根中一类特殊贮藏蛋白,该文从甘薯块根中TI组成分布、提取纯化、活性及影响因子、TI与蛋白含量相关性和TI功能性等方面对国内外有关TI研究进展进行综述。     

Trypsin Inhibitor Activity In Vitro Digestibility and Sensory Quality of Meat-Like Yuba Products as Affected by Processing

ABSTRACT: :
Soybeans ( Glycine max ) were soaked and ground to obtain soymilk. The soymilk was cooked in an open tank and held at 85 to 90 deg;C. Yuba films were picked up in 20 min intervals and dried for 20 min. Yuba films were soaked in chicken-flavor solutions (25% and 35%), and baking soda (BS) solutions (0%, 1%, 2%, and 3% BS), and cooked at 100 °C for 30 min, 60 min, and 90 min. TIA decreased (p < 0.05) with the increase of heating time and BS concentration. In vitro protein digestibility (IVPD) decreased with heating time and BS concentration (p < 0.05). Sensory characteristics were affected by flavor concentration. By using 0% BS, 25% of the chicken flavor concentration, and a short heating time method, meat-like products with low TIA, high IVPD, and good sensory characteristics were obtained.     

Effect of Ohmic Heating of Soymilk on Urease Inactivation and Kinetic Analysis in Holding Time

To verify the effect of the ohmic heating on the urease activity in the soymilk, the ohmic heating methods with the different electrical field conditions (the frequency and the voltage ranging from 50 to 10 kHz and from 160 to 220 V, respectively) were employed. The results showed that if the value of the urease activity measured with the quantitative spectrophotometry method was lower than 16.8 IU, the urease activity measured with the qualitative method was negative. The urease activity of the sample ohmically heated was significantly lower than that of the sample conventionally heated (P < 0.01) at the same target temperature. It was concluded that the electrical field enhanced the urease inactivation. In addition, the inactivation kinetics of the urease in the soymilk could be described with a biphasic model during holding time at a target temperature. Thus, it was concluded that the urease in the soymilk would contain 2 isoenzymes, one is the thermolabile fraction, the other the thermostable fraction, and that the thermostable isoenzyme could not be completely inactivated when the holding time increased, whether the soymilk was cooked with the conventional method or with the ohmic heating method. Therefore, the electric field had no effect on the inactivation of the thermostable isoenzyme of the urease.     

A simple blanch-dry process for lipoxygenase inactivation in soybean cotyledons

Full-fat flour from unblanched soybeans develops a beany flavour during milling due to lipoxygenase activity; the flour also becomes bitter during storage. The minimum thermal treatment necessary to inactivate lipoxygenase in soybeans before milling was determined. Soybeans were dehulled and cotyledons blanched in water between 80 and 99.3°C. Water blanching for 3 min at 99.3°C was necessary when blanched cotyledons were subsequently sun dried, and the flour had a nitrogen solubility index (NSI) of 37.8% and residual trypsin inhibitor activity (TIA) of 64.2%. Only 1.5 min blanching was necessary to inactivate lipoxygenase in soy cotyledons if they were dried subsequently at 60°C; NSI of this flour was 32.7%, with 44.7% residual TIA.     

Comparative Assessment of Trypsin Inhibitor vis-à-vis Kunitz Trypsin Inhibitor and Bowman-Birk Inhibitor Activities in Soybean

Trypsin inhibitor activity (TIA) in soybean is attributed to two polypeptides, namely, Kunitz trypsin inhibitor (KTI) and Bowman-Birk inhibitor (BBI). Standard spectrophotometric protocol widely followed for estimation of TIA is cumbersome and does not distinguish KTI from BBI. In the present investigation, extraction conditions for KTI were optimized and different forms of this polypeptide were resolved in 180 soybean genotypes of Indian and exotic origin through native PAGE. This led to the identification of three KTI alleles, namely, Ti^a, Ti^b, and Ti^c, with Ti^a occurring in most of the Indian genotypes. Trypsin-KTI complex assay exhibited binding of Ti^a polypeptide with 2.51 fold concentration of trypsin. Subsequently, seeds of selected genotypes were subjected to estimation of KTI and BBI activity through densitometry and enzyme-linked immunosorbent assay (ELISA), respectively; and total TIA through standard spectrophotometric protocol. Summation of KTI and BBI was significantly (P?<?0.05) lower than that of TIA determined through the spectrophotometric method.     

Effect of thermal pretreatment of raw soymilk on the gel strength and microstructure of tofu induced by microbial transglutaminase

The influence of thermal pretreatment of raw soymilk on the gel hardness and microstructure of tofu, induced by microbial transglutaminase (MTGase), was investigated in this paper. Modulated differential scanning calorimetry analysis showed that individual proteins in soymilk were to a various extent denatured by different thermal pretreatments. The viscosity of the soymilk and the gel hardness of MTGase-induced tofu were more highly related with the heating rate (up to 90 °C) than the mode of heating. At any enzyme concentration of MTGase, the tofus prepared from soymilk heated at 75 °C for 10 or 30 min showed highest gel hardness among all tested ones (P?0.05). Scanning electron microscopy analysis indicated that the microstructure of the tofu from soymilk heated at 75 °C for 30 min had a unique coral-like structure, much more continuous and homogenous than that from soymilk at 95 °C for 5 min. These results confirmed that the appropriate heat pretreatment (e.g. in the present, at 75 °C for 10-30 min) remarkably improved the gel strength of tofu by means of MTGase, and strengthened the tofu gel structure.     

Characterisation of trypsin and α-chymotrypsin inhibitors in Australian wattle seed (Acacia victoriae Bentham)

Crude water extracts from Australian wattle seed (Acacia victoriae Bentham) and their salt (ammonium sulphate)-precipitated fractions were analysed for trypsin and α-chymotrypsin (chymotrypsin) inhibitor activity, using gel electrophoresis and spectrophotometric methods. Three different bands with molecular weight 30.20, 38.03 and 39.81 kDa were active, with the 50% salt-precipitated fraction exhibiting highest activity and number of active bands. The same proteins also appeared to be responsible for both trypsin and chymotrypsin inhibitor activity. To establish conditions for the inactivation of these inhibitors, whole seed and uncoated (dehulled) cotyledon were subjected to different heat treatments. Moist heat treatment at 100 °C for 30 s was sufficient to inactivate both protease inhibitors although the trypsin inhibitor was found to be more heat-resistant than was the chymotrypsin inhibitor. Soaking overnight, before thermal treatment, improved the trypsin inhibitor activity but enhanced the efficiency of thermal inactivation in both inhibitors.     

Factors affecting the in vitro protein digestibility of chickpea albumins

In vitro protein digestibility (IVPD) of chickpea albumins and its possible relationship to their structure and the presence of trypsin inhibitor activity (TIA) have been studied. Trypsin digestion of the albumin fraction under non‐reducing conditions was incomplete, while the reduction of inter‐ and intramolecular disulphide bonds caused an improvement in the accessibility of sites susceptible to trypsin digestion. Trypsin inhibitor activity in the chickpea albumin fraction was dependent upon both temperature and heating time. Although heating the albumin fraction at 100 °C for 30 min reduced the TIA by more than 50% with respect to the initial activity, an important TIA rate was attributable to heat‐resistant trypsin inhibitor. The TIA decrease was not related to an increase in the rate of IVPD. However, we observed a significant (P ≤ 0.05) increment in IVPD in the presence of β‐ME, confirming the essential role of disulphide bonds in stabilising the protein structure of the albumin fraction. © 2000 Society of Chemical Industry     

Studies on the processing and properties of soymilk. II. Effect of processing conditions on the trypsin inhibitor activity and the digestibility in vitro of proteins in various soymilk preparations

Soymilks prepared from beans which had been soaked either in water or 0.4 M sodium carbonate solution for 24 h as a pretreatment were subjected to heat treatment under different conditions for varying lengths of time, to destroy the trypsin inhibitor activity. The rate of inactivation oftrypsia inhibitor in soy milks prepared from carbonate presoaked beans was faster than that of the water presoaked preparation when processed at 98°C and this effect was primarily associated with the change that occurred in the pH of the former system; The effect of alkaline pH's at 98°C on the inactivation of trypsin inhibitor was examined and it was found that the rate of inactivation was changed from zero order at pff 6.8 to first-order kinetics at pH 9.9. Regression equation relating pH of the system and time of heating at 98°C for 100% destruction of the inhibitor activity is presented. This effect of pretreatment was eliminated when both milks were processed at 115°C in cans because of the constancy of pH under these conditions. The influence of heat processing conditions on the enzymic digestibility of proteins in both soymilks was also studied. The pepsin digestion showed no significant differences between milks prepared from the water and the carbonate presoaked soybeans, and was highest in milks which had not been heat treated. With trypsin the digestibility increased with the degree of heal treatment up to the point where the trypsin inhibitor was destroyed, after which further heating resulted in lower digestibilities. A 19% increase in digestibility by trypsin was observed in the milk prepared from carbonate presoaked beans when compared with that from water presoaked beans and after both milks had been heated at 98°C for just sufficient time to destroy the trypsin inhibitor. When the milk prepared from water presoaked beans was autoclaved at 115°C to the same end point, its digestibility increased but it was still about 6% lower than that of the milk prepared by alkali presoaking method and processed at 98°C. The digestibility of casein using pepsin under the conditions used was lower than that of both soymilks. When trypsin was used the digestibility of casein was approximately the same as that of the adequately processed soymilk from carbonate presoaked soybeans or the autoclavtd soymilk from water presoaked beans. Other effects of using a carbonate presoaking treatment far the production of soymilk have also been discussed.     

Effect of cooking on the formation of N-nitrosodimethylamine in Korean dried seafood products

Only N-nitrosodimethylamine (NDMA) was detected when N-nitrosamines (NA) were analysed in seven dried seafood products, either uncooked or cooked. The cooking methods used were a briquet fire, a gas range, an electric oven, a microwave oven, a steam cooker and an electric coil cooker. The contents of NDMA ranged from 1.0 to 46.9 μg kg^-1 in uncooked products. When these samples were cooked, regardless of the cooking method, the content of NDMA tended to increase, ranging from 1.1 to 630.5 μg kg^-1. In general, indirect heating such as a steam cooker and a microwave oven, as compared with direct heating such as a gas range and a briquet fire, caused less increase in NDMA during cooking.     

Quality attributes of vegetable soybean as a function of boiling time and condition

Vegetable soybeans are marketed fresh or frozen, either shelled or in pods. The objective of this research was to characterise the change in quality attributes of vegetable soybean with boiling time (0–20 min), and presence/absence of pods, using an electrical-resistance stove or a steam-jacketed kettle. Trypsin inhibitor activity (TIA), texture, colour, soluble sugars, nitrogen, calcium and iron content were analysed. Blanching using a steam-jacketed kettle for approximately 2 min rendered 80% inactivation of TIA, and resulted in high colour, texture and sucrose. There were no differences between blanching in pods or shelled for TIA, colour and texture; however, blanching in pods prevented losses of sucrose. Blanching did not affect iron, mono- and oligosaccharide levels, but increased nitrogen and calcium content. Additionally, we observed that all traits decreased linearly with cooking time when using an electrical-resistance stove, except for calcium and nitrogen that increased, and oligosaccharides that remained constant.     

高静压技术对豆浆中脂肪氧合酶的钝化效果及其动力学分析

研究了高静压技术(HHP)对豆浆中脂肪氧合酶(LOX)活性的钝化作用,并进行失活动力学分析。结果表明:HHP处理能显著钝化豆浆中LOX活性。用300～600MPa压力处理5～20min,对LOX活性有显著的钝化效果(P<0.05),并且随处理压力和处理时间的增加,酶的失活率提高。当压力500MPa,保压时间0min时也有一定的钝化LOX效果;当压力600MPa处理10min以上,豆浆LOX活性被完全抑制。用500MPa处理5min,可以达到与传统巴氏杀菌相同的钝酶效果(P>0.05)。HHP技术钝化豆浆LOX的过程可用一级动力学模型拟合(R2>0.900)。随着压力的升高和处理时间的延长,k值逐渐升高,D值逐渐减小;动力学参数ZP和Va分别为125.94MPa和-45.290cm3/mol。HHP技术在钝化豆浆中脂肪氧合酶活性方面比传统巴氏杀菌彻底,效果更好。其在改善豆浆品质方面具有良好的应用前景。