Enzymatic hydrolysis of brewers' spent grain proteins and technofunctional properties of the resulting hydrolysates

Brewers' spent grain (BSG) is the insoluble residue of barley malt resulting from the manufacture of wort. Although it is the main byproduct of the brewing industry, it has received little attention as a marketable commodity and is mainly used as animal feed. Our work focuses on one of the main constituents of BSG, i.e., the proteins. The lack of solubility of BSG proteins is one of the limitations for their more extensive use in food processing. We therefore aimed to generate BSG protein hydrolysates with improved technofunctional properties. BSG protein concentrate (BPC) was prepared by alkaline extraction of BSG and subsequent acid precipitation. BPC was enzymatically hydrolyzed in a pH-stat setup by several commercially available proteases (Alcalase, Flavourzyme, and Pepsin) for different times and/or with different enzyme concentrations in order to obtain hydrolysates with different degrees of hydrolysis (DH). Physicochemical properties, such as molecular weight (MW) distribution and hydrophobicity, as well as technofunctional properties, such as solubility, color, and emulsifying and foaming properties, were determined. Enzymatic hydrolysis of BPC improved emulsion and/or foam-forming properties. However, for the hydrolysates prepared with Alcalase and Pepsin, an increasing DH generally decreased emulsifying and foam-forming capacities. Moreover, the type of enzyme impacted the resulting technofunctional properties. Hydrolysates prepared with Flavourzyme showed good technofunctional properties, independent of the DH. Physicochemical characterization of the hydrolysates indicated the importance of protein fragments with relatively high MW (exceeding 14.5 k) and high surface hydrophobicity for favorable technofunctional properties.     

Effects of ultrasound pretreatment on the enzymatic hydrolysis of soy protein isolates and on the emulsifying properties of hydrolysates

Soy protein isolate (SPI) was modified by ultrasound pretreatment (200 W, 400 W, 600 W) and controlled papain hydrolysis, and the emulsifying properties of SPIH (SPI hydrolysates) and USPIH (ultrasound pretreated SPIH) were investigated. Analysis of mean droplet sizes and creaming indices of emulsions formed by SPIH and USPIH showed that some USPIH had markedly improved emulsifying capability and emulsion stabilization against creaming during quiescent storage. Compared with control SPI and SPIH-0.58% degree of hydrolysis (DH), USPIH-400W-1.25% (USPIH pretreated under 400W sonication and hydrolyzed to 1.25% DH) was capable of forming a stable fine emulsion (d43=1.79 μm) at a lower concentration (3.0% w/v). A variety of physicochemical and interfacial properties of USPIH-400W products have been investigated in relation to DH and emulsifying properties. SDS-PAGE showed that ultrasound pretreatment could significantly improve the accessibility of some subunits (α-7S and A-11S) in soy proteins to papain hydrolysis, resulting in changes in DH, protein solubility (PS), surface hydrophobicity (H0), and secondary structure for USPIH-400W. Compared with control SPI and SPIH-0.58%, USPIH-400W-1.25% had a higher protein adsorption fraction (Fads) and a lower saturation surface load (Γsat), which is mainly due to its higher PS and random coil content, and may explain its markedly improved emulsifying capability. This study demonstrated that combined ultrasound pretreatment and controlled enzymatic hydrolysis could be an effective method for the functionality modification of globular proteins.     

Glycosylation and Deamidation of Rice Endosperm Protein for Improved Solubility and Emulsifying Properties

Rice endosperm protein was prepared by alkali-extraction method and subsequently modified by controlled glycosylation (RP_Glu, RP_XG), deamidation (RP_DA), and enzymatic hydrolysis by alcalase (RP_Alc) methods. The RP_Glu and RP_XG were prepared by Maillard type glycosylation with D-glucose and xanthan gum, respectively. The glycosylation improved the emulsion activity (0.721) and stability (26.8 min) of the protein but did not show a substantial improvement in solubility (39.7%). The rice protein modified by controlled alkali-deamidation (RP_DA) showed highest solubility (68%), emulsion activity (0.776), and emulsion stability (24 min) among the three protein modification methods evaluated in this study. The alcalase treatment to 1.8% DH (RP_Alc) slightly improved solubility (33%), emulsion activity (0.468), and emulsion stability (17.5 min) compared with unmodified rice protein (RP), which had 18% solubility, 0.266 emulsion activity, and 14.7 min emulsion stability. The glycosylation and deamidation methods were more effective than the controlled enzymatic hydrolysis by alcalase in improving solubility and emulsifying properties of rice endosperm protein. Glycosylated and deamidated rice endosperm proteins can find application in enhancing emulsifying properties in suitable products.     

高压微射流压力对大豆蛋白-大豆多糖体系功能性质的影响

为提升大豆分离蛋白（soy protein isolate,SPI）的功能性质,该文引入大豆可溶性多糖（soybean soluble polysaccharides,SSPS）,构建大豆分离蛋白-大豆可溶性多糖体系（SPI-SSPS）,研究动态高压微射流（dynamic high-pressure microfluidization,DHPM）处理对SPI-SSPS功能特性的影响。分别采用0,60,100,140和180 MPa的 DHPM压力处理SPI-SSPS,探究不同压力对SPI-SSPS起泡特性、乳化特性、溶解性、粒度分布和表面疏水性的影响。结果表明,DHPM处理能提高SPI的溶解性和起泡特性,且SSPS的存在能显著提高DHPM对SPI功能性质的改善效果（P<0.05）。100和60 MPa的DHPM处理能使SPI-SSPS呈现较高的起泡能力和起泡稳定性,分别为未处理样品的1.2和2.4倍。140 MPa的DHPM处理使SPI-SSPS溶解性较强,为未处理样品的1.8倍。然而,DHPM处理会显著降低SPI-SSPS的乳化特性、粒径和表面疏水性（P<0.05）。随着处理压力的增加,SPI-SSPS的粒度和表面疏水性逐渐降低,在180MPa的DHPM处理下SPI-SSPS具有较小的粒径和较低的荧光强度。综上所述,DHPM结合SSPS改性技术可用于改善SPI的功能性质（如溶解性、起泡性）,促进SPI在食品工业的应用。该文的研究结果可为SPI的功能性质改性提供参考。     

Emulsifying and Foaming Properties of Gluten Hydrolysates with an Increasing Degree of Hydrolysis: Role of Soluble and Insoluble Fractions

Gluten solubility was improved by enzymatic proteolysis at moderate acidic pH level. Reversed‐phase HPLC analysis of gluten hydrolysates with a degree of hydrolysis (DH) in the range of 0–5% showed that both hydrophilic and hydrophobic soluble peptides were released. Emulsifying and foaming properties of hydrolysate dispersions at 3.75 mg/mL decreased with the increasing DH at all pH levels and salt conditions investigated. On the other hand, the soluble fractions separated from those hydrolysate dispersions exhibited good functional properties, independently of the initial DH. The proportion of hydrophilic and hydrophobic peptides in the soluble fractions depended on DH, pH level, and salt concentration. Nevertheless, these soluble fractions were characterized by an excellent capacity to stabilize both oil‐water and air‐water interfaces.     

Physicochemical Properties of Rice Endosperm Proteins Extracted by Chemical and Enzymatic Methods

Rice proteins are nutritional, hypoallergenic, and healthy for human consumption. Efficient extraction with approved food‐grade enzymes and chemicals are essential for commercial production and application of rice protein as a functional ingredient. Rice endosperm proteins were isolated by alkali, salt, and enzymatic methods and evaluated for extractability and physicochemical properties. Alkali (RP_A) and salt (RP_S) methods extracted 86.9 and 87.3% of proteins with 65.9 and 58.9% yield, respectively. The enzymatic methods with Termamyl (RP_ET) and amylase S (RP_EA) extracted 85.8 and 81.0% proteins with 85.2 and 86.2% yield, respectively. Enthalpy values of RP_A (1.79 J/g), RP_S (1.22 J/g), RP_ET (nondetectable), and RP_EA (0.17 J/g), determined by differential scanning calorimetry, demonstrated that the varying level of denaturation of proteins depends on the method of extraction. Surface hydrophobicity data supported this observation. Alkali‐ and salt‐extracted proteins had higher solubility and emulsifying properties than those of enzyme‐extracted proteins. Comparatively, more favorable protein composition, lower surface hydrophobicity, higher solubility, and a lower degree of thermal denaturation of alkali‐ and salt‐extracted proteins contributed to higher emulsifying and foaming properties than those of enzyme‐extracted proteins; therefore, alkali‐ and salt‐extracted proteins can have enhanced functional use and a potential starting material for preparing tailored rice protein isolates.     

Effect of alkaline deamidation on the structure, surface hydrophobicity, and emulsifying properties of the Z19 alpha-zein

Different deamidation conditions for the Z19 alpha-zein were studied in order to find the best conditions for the development of the emulsifying properties. Alkaline deamidation was chosen, and the effects on the peptide bond cleavage, secondary structure, emulsifying properties, and surface hydrophobicity were studied. The Z19 alpha-zein was deamidated by using 0.5 N NaOH containing 70% ethanol at 70 degrees C for 12 h. A deamidation degree (DD) of 60.6 +/- 0.5%, and a degree of hydrolysis (DH) of 5 +/- 0.5% were achieved. Analysis by far-UV circular dichroism showed that the denaturation was mainly promoted by the high temperature used during the incubation. The adequate balance between the DD and the DH results in an effective emulsifying property improvement for the Z19 alpha-zein. Thus, after the deamidation treatment, the surface hydrophobicity decreased from 9.5 x 104 +/- 6.8 x 103 to 46 x 104 +/- 2.1 x 103, and the emulsion stability increased from 18 +/- 0.7% to 80 +/- 4.7% since the oil globules stabilized by the modified protein were smaller (57.7 +/- 5.73 nm) and more resistant to coalescence than those present in the native protein emulsions (1488 +/- 3.92 nm).     

Functional Properties of Rice Bran Protein Isolate at Different pH Levels

The functional properties of proteins from Tarom and Shiroodi cultivars were determined and compared with technological aspects of food and nutraceutical applications. Shiroodi has higher protein content than Tarom, and the yields of protein obtained were 72.88 and 66.36%, respectively. Nitrogen solubilities of rice bran protein of Tarom were more than Shiroodi at all pH levels. In addition, higher solubility was found in acidic or alkaline conditions. Although the rice bran proteins had lower emulsifying properties than bovine serum albumin, they had similar foaming properties in comparison with egg white. Tarom isolates had a significantly higher solubility, emulsifying property, and foaming stability and greater surface properties than Shiroodi isolates. The results showed the surface hydrophobicities of rice bran protein were greater than casein and ovalbumin and lower than other proteins such as bovine serum albumin. Water and oil absorption capacities were 1.03 and 1.66 for Tarom and 87.3 and 75.3 for Shiroodi, respectively. The bulk densities of Tarom and Shiroodi were also 0.55 and 0.53 g/mL, which make them suitable for weaning food and other industrial applications. As a result, these rice bran proteins showed higher hydrophobicity than that of other rice bran protein varieties as well as more functionality. Thus, they have good potential in the food and pharmaceutical industries.     

beta-lactoglobulin hydrolysis. 1. Peptide composition and functional properties of hydrolysates obtained by the action of plasmin, trypsin, and Staphylococcus aureus V8 protease.

beta-Lactoglobulin (betaLg) was subjected to limited hydrolysis by trypsin, plasmin, and endoproteinase from Staphylococcus aureus V8 (S.aur.V8) to degrees of hydrolysis (DH) of 1, 2, and 4%. The several hydrolysates had different peptide compositions (determined by reversed-phase HPLC and gel-permeation chromatography [GPC]). GPC under nondenaturing, denaturing, and denaturing plus reducing conditions showed that the peptides formed were linked by hydrophobic interactions or by disulfide bonds or were not linked at all. At very low protein concentration, some differences in emulsion-forming properties were observed: only the plasmin hydrolysates could form emulsions with a uniform particle-size distribution. The emulsions formed with S.aur.V8 hydrolysates had poor emulsion-stabilizing properties. Some hydrolysates showed increased foam-forming properties in comparison with the intact protein. All foams formed were stable. Overall, the plasmin hydrolysate (DH4) contained relatively much larger molecules and/or hydrophobic molecules. Many molecules were disulfide-linked peptides. This hydrolysate also had the best functional properties.     

Preparation of Rice Endosperm Protein Isolate by Alkali Extraction

Rice endosperm extraction conditions were optimized by response surface methodology. The optimum alkali extraction conditions were pH 11.0 at 40°C for 3 hr with 8:1 solvent‐to‐solid ratio. The maximum protein yield was 43.1% at these conditions. As the extraction pH was increased from 9.0 to 12.0, protein extractability and content increased but the solubility and emulsifying properties of the extracted protein decreased. The extracted protein was recovered by either isoelectric precipitation (IEP) or ultrafiltration (UF). Ultrafiltering the supernatant with a 5‐kDa hollow fiber membrane concentrated the protein from 1.8 to 16% (dry basis) and the resulting solution was spray‐dried to produce a protein concentrate (RP_UF) with 71% protein. Although RP_IEP contained higher protein (86%) than RP_UF (71%), RP_UF showed higher solubility and emulsifying properties. The solubility of RP_UF was higher (37%) than RP_IEP (15%). RP_UF also demonstrated higher emulsion activity (0.414) and stability (22.4 min) compared with the emulsion activity (0.282) and stability (15.5 min) of RP_IEP. Higher solubility and the soluble nonprotein components of RP_UF contributed to higher emulsifying properties than RP_IEP. The UF provided milder extraction conditions with improved emulsifying properties than conventional IEP.     

Optimization of the enzymatic deamidation of soy protein by protein-glutaminase and its effect on the functional properties of the protein

The effects of enzymatic deamidation by protein-glutaminase (PG) on the functional properties of soy protein isolate (SPI) were studied. Conditions for the deamidation were evaluated by means of response surface methodology (RSM). Optimal conditions based on achieving a high degree of deamidation (DD) with a concurrently low degree of hydrolysis (DH) were 44 °C, enzyme:substrate ratio (E/S) of 40 U/g protein and pH 7.0. Under optimal conditions, both DD and DH increased over time. SDS-PAGE results indicated that lower molecular mass subunits were produced with increasing DD. Far-UV circular dichroism spectra revealed that the α-helix structure decreased with higher DD, while the β-sheet structure increased until 15 min of deamidation (32.9% DD), but then decreased at higher DD. The solubility of deamidated SPI was enhanced under both acidic and neutral conditions. SPI with higher DD showed better emulsifying properties and greater foaming capacity than SPI, while foaming stability was decreased. It is possible to modify and potentially improve the functional properties of SPI by enzymatic deamidation using PG.     

Preparation and characterization of papain-modified sesame (Sesamum indicum L.) protein isolates

Defatted sesame meal ( approximately 40-50% protein content) is very important as a protein source for human consumption due to the presence of sulfur-containing amino acids, mainly methionine. Sesame protein isolate (SPI) is produced from dehulled, defatted sesame meal and used as a starting material to produce protein hydrolysate by papain. Protein solubility at different pH values, emulsifying properties in terms of emulsion activity index (EAI) and emulsion stability index (ESI), foaming properties in terms of foam capacity (FC) and foam stability (FS), and molecular weight distribution of the SPI hydrolysates were investigated. Within 10 min of hydrolysis, the maximum cleavage of peptide bonds occurred as observed from the degree of hydrolysis. Protein hydrolysates have better functional properties than the original SPI. Significant increase in protein solubility, EAI, and ESI were observed. The greatest increase in solubility was observed between pH 5.0 and 7.0. The molecular weight of the hydrolysates was also reduced significantly during hydrolysis. These improved functional properties of different protein hydrolysates would make them useful products, especially in the food, pharmaceutical, and related industries.     

Functionality of beta-casein peptides: importance of amphipathicity for emulsion-stabilizing properties.

To investigate structure-function relationships with regard to emulsion-stabilizing properties, peptides from bovine beta-casein (betaCN), obtained by plasmin hydrolysis and fractionation of the hydrolysate, were isolated and identified on the basis of their masses determined by electrospray ionization mass spectrometry, the primary structure of the intact protein, and the known specificity of the enzyme. An amphipathic peptide fraction was fractionated further by ion-exchange chromatography and subsequent hydrophobic interaction chromatography resulting in the components betaCN[f 1-105/107] and betaCN[f 29-105/107]. The latter peptides had poor emulsion-stabilizing properties compared to the former ones, and the stability of an emulsion formed with betaCN[f 29-105/107] was also more sensitive to hydrophobic impurities than that of an emulsion formed with betaCN[f 1-105/107]. The highly charged N-terminal part appeared to be important for the emulsion-stabilizing properties of these peptides. A hypothesis for the structure-function relationship is given.     

脱酰胺与双酶协同作用提高小麦面筋蛋白酶解效率

为了探讨了不同脱酰胺处理和双酶协同作用方式对小麦面筋蛋白酶解效率及其产物抗氧化活性的影响,该文研究了小麦面筋蛋白在各种预处理方式和酶解条件下的蛋白回收率、水解度、抗氧化性能及肽分子量分布情况。结果显示,单独热处理(90℃,30 min)小麦面筋蛋白对其酶解效率无显著影响,而采用添加0.5 mol/L柠檬酸溶液进行热处理(质量分数为5%,90℃,30 min)可显著(P0.05)提高其蛋白回收率。此外,酶制剂添加顺序及双酶共同水解作用时间对酶解效率均具有较大影响:加入谷氨酰胺酶预先水解对小麦面筋蛋白的深度水解有促进作用;一定时间内的双酶协同作用有利于酶解的进行,但较长时间的双酶作用反而会抑制酶解效率。采用谷氨酰胺酶(质量分数为0.2%)对经柠檬酸加热处理的小麦面筋蛋白作用12 h后再加入胰酶(质量分数为0.6%)共同作用7 h可使蛋白回收率达70.74%,水解度达到9.88%;另外,酶解产物的自由基清除能力ABTS+(2,2’-Azinobis-(3-ethylbenzthiazoline-6-sulphonate)+)值与氧化自由基吸收能力(ORAC,oxygen radical absorbance capacity)值分别达到478.95 mmol/g和213.85μmol/g,提示该酶解产物是一种潜在优秀食品抗氧化剂。研究结果可为拓宽小麦面筋蛋白的应用领域,以及高效制备抗氧化活性肽提供方法和理论指导。     

燕麦麸分离蛋白的酶解对其功能性质的影响

为了改善燕麦蛋白的功能性质以扩大其在食品工业中的应用,该文以燕麦麸为原料制备了燕麦麸分离蛋白(OBPI),并利用胰蛋白酶对其进行水解,得到了3种不同水解度(4.1%、6.4%、8.3%)的酶解产物。SDS-PAGE分析结果表明OBPI中的主要蛋白成分是球蛋白,其经过胰蛋白酶处理后,球蛋白酸性亚基被部分水解而碱性亚基相对保持完整。胰蛋白酶水解显著改变了OBPI的功能性质。在所考察的水解度范围内,随着水解度的升高,酶解产物的溶解性、持水性、乳化活性及起泡能力等方面均逐渐增加;但持油性、乳化及泡沫稳定性有不同程度的降低。     

Functional Properties of 7s Globulin Extracted from Cowpea Vicilins

The seed of cowpea (Vigna unguiculata L.) is rich in protein and the amino acid profiles of the meal are suitable for human dietary products, but little is known about the structure and chemical properties of the protein extracted from this legume. This study determined the functional properties of two selected cowpea cultivars and their solubility, emulsifying capacity, surface hydrophobicity, and thermal stability. Seeds of red and black cowpea were surface sterilized and the 7s globulin was isolated and purified using column chromatography with Sephacryl S‐300 (Hi‐Prep 26/60) gel column. Also, SDS‐PAGE and protein structure were analyzed using biochemical procedures. At high ionic strength (μ = 0.5), cowpea 7s globulin fraction exhibited better solubility for a wide range of pH levels, higher emulsifying capacities, and greater thermal stability than those obtained at low ionic strength (μ = 0.08). The lowest solubility was observed at pH 5.3–6.4 at the low ionic strength. Emulsifying capacities at high protein concentration were greater when compared with low protein concentration. T_m values of black cowpea globulin fraction were higher than those of red cowpea globulin fraction, whereas the surface hydrophobicity of the globulin fraction in red cowpea was larger than that in black cowpea.     

Heat-induced destabilization of oil-in-water emulsions formed from hydrolyzed whey protein.

The emulsifying ability, heat stability, and coalescence stability of oil-in-water emulsions prepared with whey protein of varied degrees of hydrolysis (DH), and at varied protein contents, was studied. Whey protein hydrolysates (WPH) with a DH of 4% and 10% had poorer emulsifying ability than non-hydrolyzed whey protein concentrate (WPC), but were more heat stable. Increasing DH between 10 and 27% improved emulsifying ability and further improved the heat stability of the emulsion droplets. Increasing DH from 27 to 35% led to a big decrease in both emulsifying ability and heat stability. The quiescent coalescence stability of WPH emulsions was relatively good up to a DH of 27%. Above DH 27% emulsions become highly unstable. It appears that two mechanisms of instability are at work here. At low DH heat-induced denaturation and aggregation occur. In the DH range of 4-20% heat stability increases as protein globular structure is disrupted. At a DH greater than 27% we see a change from a hydrolysis-induced increase in heat-stability to coalescence instability, with a resultant large increase in emulsion breakdown during heating.     

Perfusion chromatography purification of a 15 kDa rice prolamin

Prolamin extracted from rice flour using 55% n-propanol contained protein impurities. Reverse phase high-performance liquid chromatography (HPLC) on a perfusion column R2/H was used to separate rice prolamin from other proteins in less than 5 min. Prolamin eluted as the major peak. The isolated prolamin migrated as a single band on sodium dodecyl sulfate polyacrylamide gel electrophoresis using a 4-12% Bis-Tris gel. Matrix-assisted laser desorption ionization mass spectrometry identified the rice prolamin as a 15 013 Da protein. The surface hydrophobicity (S(o)) of the HPLC-separated protein fractions was measured using the hydrophobic fluorescent probe PRODAN. A comparison was made with the surface hydrophobicity (S(o)) of corn prolamin and bovine serum albumin. Surface hydrophobicity values and solubility in 90% ethanol assisted in rice prolamin identification from other chromatographic peaks. The advantage of perfusion chromatography in purifying rice prolamin from other rice proteins included the reduced separation time, the speed at which the separation was carried, and the ability to regenerate the column in a short period of time and allow for more samples to be purified and separated.     

Effect of enzymatic treatment of extracted sunflower proteins on solubility, amino acid composition, and surface activity

Industrial proteins from agriculture of either animal or vegetable origin, including their peptide derivatives, are of great importance, from the qualitative and quantitative point of view, in food formulations (emulsions and foams). A fundamental understanding of the physical, chemical, and functional properties of these proteins is essential if the performance of proteins in foods is to be improved and if underutilized proteins, such as plant proteins (and their hydrolysates and peptides derivatives), are to be increasingly used in traditional and new processed food products (safe, high-quality, health foods with good nutritional value). In this contribution we have determined the main physicochemical characteristics (solubility, composition, and analysis of amino acids) of a sunflower protein isolate (SPI) and its hydrolysates with low (5.62%), medium (23.5%), and high (46.3%) degrees of hydrolysis. The hydrolysates were obtained by enzymatic treatment with Alcalase 2.4 L for DH 5.62 and 23.5% and with Alcalase 2.4 L and Flavorzyme 1000 MG sequentially for DH 46.3%. The protein concentration dependence on surface pressure (surface pressure isotherm), a measure of the surface activity of the products (SPI and its hydrolysates), was obtained by tensiometry. We have observed that the degree of hydrolysis has an effect on solubility, composition, and content of the amino acids of the SPI and its hydrolysates. The superficial activity and the adsorption efficiency were also affected by the degree of hydrolysis.     

Isolation, purification, and identification of protein associated with corn fiber gum

Corn fiber gum (CFG), an alkaline hydrogen peroxide extract of the corn kernel milling byproduct "corn fiber", is a proteinaceous arabinoxylan with protein content ranging from ca. 2 to 9% by weight for CFG samples isolated from different corn milling fiber sources. Several studies have suggested that protein associated with CFG could be partly responsible for its excellent emulsifying properties in oil-in-water emulsion systems. Nevertheless, the composition and identity of the protein component has never been determined. In the present study, CFG was deglycosylated by treating with trifluoromethanesulfonic acid, and the resulting proteins were purified by passage through C18 solid phase extraction cartridges. The proteins were then separated and characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The protein band from the gel was treated with a proteolytic enzyme, chymotrypsin, and the resulting peptides were cleaned using C18 Zip Tip pipet tips and analyzed using matrix-assisted laser desorption/ionization with automated tandem time-of-flight mass spectrometry. The partial sequences derived from the mass spectrometry analyses of the resulting chymotryptic peptides were found to be similar to the 22-kDa alpha-zein Z1 (az22z1) protein (a major storage protein in corn endosperm) when queried against the primary sequences from the National Center for Biotechnology Information database. This is the first report that this hydrophobic protein is associated with CFG and may explain why CFG is an excellent emulsifier for oil-in-water emulsion systems.