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Comparison properties of rice bran protein hydrolyzate obtained by spray dry & freeze dry

Physicochemical Properties and Functionality of Rice Bran Protein Hydrolyzate Prepared from Heat-stabilized Defatted Rice Bran with the Aid of Enzymes
S. TANG , N.S. H ETTIARACHCHY , R. H ORAX, AND S. E SWARANANDAM

Food Chemistry and Toxicology

ABSTRACT: Molecular size, thermal properties, hydrophobicity, nitrogen solubility, and emulsifying and foaming properties were determined for protein products from heat-stabilized defatted rice bran. The freeze-dried and spray-dried proteins had molecular sizes between 6.5 to 66.2 kDa; denaturation temperatures of 84.1 and 84.6 °C, enthalpies of 2.5 and 2.37 J/g, hydrophobicities of 20677 and 22611, maximum solubilities of 66.3% and 66.1% at pH 12.0, emulsifying capacities of 0.19 and 0.18, emulsion stabilities of 16.5 and 17.3 min, foam capacities of 4.0 mL and 4.2 mL, and negligible foam stabilities. These results demonstrated that the extracted rice bran protein has potential as a nutraceutical ingredient in food applications. Keywords: rice bran protein, hydrophobicity, DSC, functionality

Introduction
because of its unique nutritional value and nutraceutical properties (Saunders 1990). Its nutritional value is much higher than rice endosperm protein or protein from any other cereals or legumes (Juliano 1994). Rice bran protein also is highly digestible. In addition to the high nutritional value, rice protein is a hypoallergenic food ingredient and may serve in infant formulations (Helm and Burks 1996). Furthermore, rice bran protein has been reported to have anti-cancer properties (Kawamura and Muramoto 1993). Knowledge on its functional properties is needed for this ingredient’s use in a range of food applications. Protein functional properties are determined to a large extent by a protein’s physicochemical and structural properties (Damodaran 1990). Protein solubility is an important prerequisite for food protein functional properties, and it is a good index of potential applications of proteins (Kinsella 1976). Researchers have reported that protein solubility has a close relationship with emulsifying (Yatsumatsu and others 1972a,b; McWatters and Holmes 1979) and foaming properties (Yatsumatsu and others 1972a). Amino acid composition and molecular size are the foundations of protein functionality (Kinsella 1981 ). Differential scanning calorimetry (DSC) has been used to study the thermal properties and the structural changes of proteins (Hermansson 1978; Patel and others 1990; Reaker and Johnson 1995). Surface hydrophobicity has also been reported to correlate to surface tension, interfacial tension, and emulsifying activity (Nakai and others 1980; Halling 1981; Petruccelli and Anon 1994; Phillips and others 1994). Bera and Mukherjee (1989) showed that rice bran protein concentrate from alkali extraction had higher nitrogen solubility at higher or lower pH values than its isoelectric point (pH 4.5). Wang and others (1999) evaluated the solubility of rice bran protein isolate in the pH range of 2.0 to 12.0 with varying concentrations of NaCl. These researchers found that the nitrogen solubility of rice bran protein isolate was between 70.0 and 90.0% in the broad pH range of 6.0 to 12.0. Rice bran protein

R

ICE BRAN PROTEIN IS GAINING INTEREST IN THE FOOD INDUSTRY

isolate had the lowest solubility (10%) at pH 4.0. Hamada (2000) reported that protein hydrolysate of protease treatment from defatted nonheat-stabilized rice bran had very good nitrogen solubility. Wang and others (1999) with SDS-PAGE and Hamada (1997, 2000) with size-exclusion HPLC reported the molecular sizes of rice bran protein from nonheat-stabilized rice bran. Wang and others (1999) reported that the denaturation temperature and enthalpy changes were 83.4 °C and 0.96 J/g for rice bran protein isolate. These investigators also reported that the hydrophobicity values of rice bran protein isolate (obtained after treating with phytase and xylanase) and rice bran concentrate without enzyme treatment were 14.0 and 20.4, respectively. Bae and Jang (1999) reported that succinylated rice bran protein concentrate had good functional properties, including solubility, emulsion properties, and oil absorption capacity. Wang and others (1999) reported emulsifying capacity and emulsion stability were 0.3 to 0.37 and 3.9 to 4.2 min, and foam capacity and stability were 0.3 to 0.5 and 4.1 to 4.3 min, respectively, for rice bran protein isolate prepared from nonstabilized rice bran. In addition to the characteristics and functional properties, Connor and others (1976) and Wang and others (1999) reported on the amino acid composition of rice bran proteins. Heat processing during oil extraction denatures rice bran protein, which could lead to higher hydrophobicity and affect functional properties. Information about molecular size, hydophobicity, nitrogen solubility, and thermal and functional properties of proteins from heat-stabilized defatted rice bran are not available in the published literature. Commercially, the oil is extracted from rice bran. During the oil extraction process, rice bran goes through a heat stabilization step to inactivate the lipase enzyme to preserve the quality of oil. The rice bran after its oil removal with heat treatment is a co-product and is designated as heat-stabilized defatted rice bran. Protein extracted from heat-stabilized defatted rice bran could be utilized as a nutraceutical food ingredient. The objectives of this study were to characterize freeze-dried (FD-RBP) and spraydried rice bran protein (SD-RBP) hydrolyzate with pectinase and

and the slurry was incubated at 45 °C and shaken at 200 rpm in a Micro-Environ shaker (Melrose Park. 3 mg of protein was mixed with 0. and dried in a Bio-Rad Model 543 Gel Dryer (Bio-Rad Laboratories). phosphorylase B (97. the same amount of protein was added with 0. The gels were run in an electrode buffer (pH 8. Electrophoresis separation of proteins Electrophoresis of FD-RBP. and to investigate their amino acid compositions.3 kDa).5-␮L volumes and run on a Waters HPLC (Waters Corporation. Soy protein isolate contain 90% protein and pH is about 7.) at 570 nm wavelength and the amino acids were quantified by comparing with standard amino acid profiles. Mo.5%. The residue of pectinase treatment was further treated with 60000 units of Protease P at pH 7.) at a flow rate of 0.5 mL nonreducing buffer.. The nitrogen content of the supernatant was determined by the Kjeldahl method and percent nitrogen solubility was calculated as follows: amount of nitrogen in supernatant × 100 nitrogen solubility. % = total amount of nitrogen in 100 mg sample Protein content Protein contents in rice bran and extracted rice bran protein products were determined by the automated Kjel-Foss method 4608 (AACC 1990). Calif..A. For reduced conditions. Mountain View.. Inc. Nitrogen solubility Nitrogen solubility was determined according to the procedure of Bera and Mukherjee (1989).A.1 M HCl or 0.A.750 units of pectinase. The combined supernatants were freeze-dried (FD-RBP) and spray-dried (SD-RBP). Calif. The supernatants from pectinase and protease treatments were combined.3) containing 0. Commercial food-grade enzymes.Physicochemical properties and functionality of rice bran protein… protease. Nonreduced/reduced protein solution (15 ␮L) was loaded onto the gel wells.4 kDa). Ark.40). U. U.2) was added and transferred to a 1..0 in 1:10 dispersion in water. The samples were heated at 95 °C for 40 min and cooled to room temperature.15.40).) for 30 min at room temperature (approximately 25 °C) and centrifuged at 4000 × g for 30 min. Milford. Calif.S. U. Materials and Methods A SINGLE BATCH OF HEAT-STABILIZED DEFATTED RICE BRAN (HDRB) was obtained from Riceland Foods. Ill.. or SPI protein was dispersed in 10 mL of distilled deionized water. The rice bran protein extract and SPI varied in protein content were used at the same concentration for evaluation of functional properties. thermal properties. U.. and SPI samples. The sealed samples were taken to a drying oven to be hydrolyzed at 120 °C for 16 h. The gels were fixed and stained with 0.S.0. then cooled and centrifuged at 1100 × g for 20 min in a centrifuge (model J2-21. One mL of this standard was diluted by 9 ␮L of nonreducing buffer and loaded to the well.A.). of nonreducing buffer and 0. lysozyme (14.2-␮L Gelman membrane filter. The air was removed by keeping the sample in the vacuum chamber.) and Sigma Chemical Co.025 mL 2-mercaptoethanol. Percent total protein content in rice bran and extracted products were read from the output of the instrument using a conversion factor of 5.) for 2.15 and 7.S.. Calif.).S. U. ovalbumin (45 kDa).S. Inc. molecular sizes. Inc (Stuttgart. (Decatur.3% (w/v) Tris base. and 5.5. The light absorbance of amino acids was detected with a UVVisible detector (Pickering Laboratories Inc.S. 10. The treated product was heated at 95 °C for 2 min to inactivate the enzymes.44% (w/v) glycine.5 kDa). and 0.8)..0 using either 0. U... Sodium ion exchange column was used to separate amino acids (Pickering Laboratories.A.0/10. Beckman. then destained with 10% acetic acid and 40% ethanol. Ill.A. 12. ␤-galactosidase (116. 5 mL of 2 mM norleucine internal standard was added and the solution passed through a 0. sodium sample diluent (pH 2.1 mL 10% (w/v) sodium dodecyl sulfate (SDS).8 and 32. hydrophobicity.S.3 mL 1% (w/v) bromophenol blue.30a (1990) was used for amino acid analysis of FD-RBP. and protein contents were determined with KjelTech Analyzer 2000 (Tecator Co. The pH was adjusted to 3. Hoganas.). Pickering Laboratories.4 mL/min with a Pickering sodium ion-exchange column of 4 × 150 mm (Pickering Laboratories. Melrose Park. Japan).). SPI was included for comparison. These suspensions were shaken (Lab-Line Environ-Shaker. trypsin inhibitor (21.5 kDa). To the freeze-dried residue. and SPI was conducted using a 12% separating gel with a 4% stacking gel (Laemmli 1970) under reducing and nonreducing conditions. One milliliter of stock sample was pipetted into a 50-mL borosilicate glass serum bottle and dried in a freeze-drier.20) and TRIONE ninhydrin reagent were purchased from Pickering Laboratories.). For nonreducing conditions.4 kDa).A.) for 41 min. Samples were digested with Kjeldahl Digestion System 6 for 1 h at 420 °C.5 mL deionized water. Electrophoresis reagents were purchased from Bio-Rad Laboratories (Hercules. SD-BRB. Mountain View. For each 100 mg of sample. Soy protein isolate (PRO FAM) was obtained from Archer Daniels Midland Co. One hundred mg of FD-RBP. (St.. which consisted of 50. Other analytical grade chemicals were purchased from Fisher Scientific (Pittsburgh. and myosin (200 kDa).S.1% (w/ v) SDS with a Bio-Rad vertical slab gel electrophoresis unit model Mini-ProteanTM II and power supply model 3000 XI (Bio-Rad Laboratory. 1. SD-RBP. Reagents for amino acid analysis sodium eluant (pH 3. Seventy milligrams of FD-RBP. Eighty percent of the total protein in rice bran was in combined supernatant.1% Coomassie Brilliant Blue R-250 in 10% acetic acid and 40% ethanol. Inc.95 (Juliano 1994).5 M Tris-HCl (pH 6.). 45 °C. The molecular mass standards from Bio-Rad was composed of aprotinin (6. 5 mL 6 M HCl was added in a 50-mL stopper bottle and sealed. Lab-Line Instrument. The suspensions were adjusted to pH 2.1 M NaOH. Louis. Sweden ).2 kDa).5 mL glycerol.S. carbonic anhydrase (31 kDa). These proteins were used for investigating molecular size. and emulsifying and foaming properties. Richmond. (Nagoya.0/4. TRIONE® ninhydrin reagent was added with post column instrument (TRIONE Ninhydrin derivatization instrument.A. Pa. and functional properties. Soy protein isolate (SPI) was included for comparison.. Inc. U.). Calif. respectively. Amino acid composition A modified AOAC method 982. Inc. U.1 h.. The protein contents in FD-RBP and SD-RBP were 32. U. and shaken at 200 rpm for 3.475 mL Differential scanning calorimetry Thermal properties of rice bran proteins were evaluated using differential scanning calorimetry (DSC). amino acid composition.6 mL 0. pectinase II (3500 units/g) and Protease P (600000 units/g) were obtained from Amano Pharmaceutical Co. serum albumin (66.S. After hydrolysis.0/6. SDBRB. Fullerton.5 h with 8.A..S. 21.. Preparation of rice bran protein Rice bran (50 g) was dispersed in 500 g deionized water. U. The prepared samples were injected as 2. Ill. SD-RBP. All chemical used were of reagent grade.0/8. pH 7. 1 mL of sodium diluent buffer (pH 2. Mass.A.0/ 12. U.) and sodium eluent (pH 3.5-mL micro-centrifuge tube for HPLC analysis. or SPI was dis- Food Chemistry and Toxicology .A.

The spray-dried and freezedried protein products were used for investigation of molecular size.01 M phosphate buffer (pH 7. This protein solution (45 ␮L) was transferred and hermetically sealed in a stainless steel pan. Results and Discussion Protein contents of isoelectrically precipitated protein and FD-RBP and SD-RBP The protein content of heat-stabilized defatted rice bran (HDRB) was 17. Foam stability (FS) was calculated from the following equation: FS=Vo × ⌬t/⌬V where ⌬V is the decrease in volume of foam during the time interval of ⌬t (15 min).A. 0. Kontron Ltd. Absorbance measured soon after homogenizing was expressed as emulsifying activity of protein.012.006.2 kDa. Two higher-density protein bands were observed between 6.1 M NaCl.05. respectively. Fluorescence intensity of these solutions was measured at 390 nm of excitation and 484 nm emmision using a Kontron Spectrofluorometer (model SFM23/ B.A. but FD-RBP had slightly higher amino acid contents than the SD-RBP. Statistical analysis Data were analyzed for variance and multiple mean comparison Figure 1—Electrophoretogram of rice bran protein from heat-stabilized defatted rice bran.1% protein solution (w/v.S.0) containing 0. The significance of differences between means was determined by the Tukey-Kramer HSD procedure at p < 0. Conn. Zurich. nitrogen solubility.0) and vortexed homogeneously.01 M phosphate buffer (pH 7. and SPI are shown in Table 1.05 M phosphate buffer (pH 7.) and denaturation peak temperature and enthalpy were calculated by a thermal analysis data software program (Pyris-I-DSC.. Ten ␮L of 8 mM ANS in 0.5% protein. The molecular weights of all 4 components were between 6.0 mL of the protein solutions. U.4) in a glass tube (2..1% SDS (w/v) at 0 and 10 min after homogenizing.4 kDa. and emulsifying and foaming properties.4 x 30 cm) for 15 s.). SRP and SRPR: nonreduced and reduced spraydried rice bran protein. Emulsifying activity and emulsion stability Emulsifying activity (EA) and emulsion stability (ES) of rice bran proteins and SPI were determined by the turbidimetric methods of Pearce and Kinsella (1978). w/v) were prepared in 0. Wang and others (1999) reported that rice bran protein isolate had more than 12 bands. thermal properties. The surface hydrophobicity. pH 7.A. was calculated by linear regression. and emulsion stability was determined as follows: ES = To × ⌬t A⌬ where ⌬T is the decrease in turbidity (absorbance) of To in the time interval ⌬t and To is the absorbance of the emulsion after homogenization. then mixed well by vortexing (setting at 5) for 10 s. SD-RBP.02 (2000) software package (SAS.Physicochemical properties and functionality of rice bran protein… solved in 1 mL of 0. and SPI were determined by using the fluorescence 1-anilino-8-naphthalene sulfonate (ANS) binding method (Hayakawa and Nakai 1985).015%. Amino acid composition The amino acid compositions of FD-RBP. Determination of surface hydrophobicity Surface hydrophobicity of FD-RBP. of the treatments with JMP 4. The Virtis Co. ST: standard (kDa). plotted as the slope of fluorescence intensity against protein concentration. U. 0.S.0) was added into each of 4.4 to 97.). amino acid composition. Norwalk. SD-BRB.0015. STR: reduced standard. Conn. Perkin-Elmer Corp. and SPI are shown in Figure 1.003.5 and 14. Rice bran protein solutions (0. The SPI had more than 10 bands and their molecular sizes ranged from 14. Cary.2% protein in 0. . 0. Switzerland). U. Gardiner.. The volume of foams was measured immediately for foam capacity.. and Vo is the volume of foam at 0 time (beginning after air introduction was stopped).A.S. In comparison with SPI. Air (90 cm3/ min) was introduced into 5 mL of 0. N. The solution was mixed and the absorbance of the SDS solution was measured at 500 nm (Varian series 634 double-beam spectrophotometer)..S.5 and 66.0) and 2 mL of soy oil were homogenized for 1 min using a Virtis homogenizer at 27000 × g (Virtishear Tempest. It is possible that protease hydrolyzed some higher-molecular-weight protein fraction into smaller-molecular-weight peptides that ran off the gel.Y. Six milliliters of 0. U. The sample was heated by scanning from 25 to 135 °C at a rate of 10 °C per min against a reference containing 45 ␮L buffer without protein in a differential scanning calorimeter (Perkin-Elmer Corp. 0. SD-RBP.. The nonreducing and reducing samples showed that the rice bran protein products had 4 components of protein. FRP and FRPR: nonreduced and reduced freeze-dried rice bran protein. N.8 and 32.2 kDa. Norwalk. Food Chemistry and Toxicology Electrophoresis of proteins The electrophoresis patterns of FD-RBP. The largest molecular size was between 45.0 and 66. hydrophobicity.) to produce the emulsion. both FD-RBP and SD- Foam capacity and foam stability Foam capacity (FC) and foam stability (FS) of rice bran protein was measured by the method of Kato and others (1983).6%. SPI and SPIR: nonreduced and reduced soy protein isolate. The FD-RBP and SD-RBP had very similar amino acid patterns.05 M phosphate buffer (pH 7.C. Freeze-dried and spray-dried rice bran protein products from supernatants with 80% protein recovered contained 32.. Portions of this emulsion (50 ␮L) were pipetted into 5 mL of 0..5 kDa.

226. SD-RBP.8 5.5 8.3 10.1 6. no significant differences were observed between FD-RBP and SD-RBP in either emulsifying activity or emulsion stability (p > 0.2 SD-RBP 10.7 5.8* 3.0. spray-dried rice bran protein (SD-RBP). and reported hydrophobicity values of 14. the solubility of SPI reached 82%.0. all 3 sources of protein had lower solubilities.77. spray-dried rice bran protein (SD-RBP). SD-RBP.8 FD-RBP 11.7 7. Wang and others (1999) reported that the denaturation temperature and enthalpy changes were 83.6 4.0 are presented in Figure 2. respectively.6 5. Emulsifying activity and emulsion stability of proteins The emulsifying activities and emulsion stabilities of FD-RBP. and 323.0 to 12. and SPI are given in Table 2.8 Table 2—Thermal properties of freeze-dried/spray-dried rice bran protein (FD-RBP/SD-RBP) from heat-stabilized defatted rice bran and soy protein isolate (SPI) Protein source Onset FD-RBP** SD-RBP SPI 67. Food Chemistry and Toxicology . Thermal properties of proteins RBP contained higher amounts of the essential amino acids threonine.2 5. the surface hydrophobicity value for SPI was significantly higher than either for FD-RBP or SD-RBP (p < 0.0 4. valine.0. Both FD-RBP and SD-RBP had higher hydrophobicities than protein isolate and concentrate of full-fat nonstabilized rice bran and this also could be due to hydrophobic amino acids exposed during heat-stabilization. and SPI at pH 2.85 DH J/g 2. However.Physicochemical properties and functionality of rice bran protein… Table 1—Amino acid composition of freeze-dried (FD-RBP).2 90. soy protein isolate (SPI) and freeze-dried and spray-dried rice bran protein products had similar solubilities (below 30%). respectively).03.7 7. the nitrogen solubilities increased rapidly with an increase in pH up to pH 12.9 89. histidine. Gnanasambandam and Hettiarachchy (1995).5 and 2. FD-RBP. the hydrophobic amino acids are buried in the central core of the protein molecule. In this study. and SPI were 206. 9. respectively. phenylalanine. The lysine and threonine contents in FD-RPB and SD-RBP agreed with the data reported by Connor and others (1976). In the native proteins.5 18. °C Peak End Area. but arginine.4 90.7 4. This feature is lost when protein is denatured or hydrolyzed into shorter peptides. and SPI had similar denaturation temperatures (84. From the whole amino acid profile.5 4. and soy protein isolate (SPI). However.7% for SPI.1 5. SPI had significantly higher emulsifying activity and emulsion stability than both FD-RBP and SD-RBP (p < 0. At pH 4.2 7.0. and valine contents contents in FD-RPB and SD-RBP were higher.5 17. while solubility for both rice bran protein products was only 66%. Wang and others (1999) reported a much lower value of lysine content (3.6. At pH 12.1 9.8* 73. and SPI are shown in Table 3.0 5.1 6. These trends in solubilities are in agreement with the data reported for rice bran pro- Surface hydrophobicity of proteins The surface hydrophobicities (So) of freeze-dried rice bran protein (FD-RBP).0.5 7.0 and 20.47 5.0.0/12.2 °C.11. The enthalpies of FD-RBP and SD-RBP were 2.4 8. tyrosine.7 4.86 *Values are means of duplicate analyses **FD-RBP and SD-RBP: freeze-dried and spray-dried rice bran protein SPI: soy protein isolate *Values are means of duplicate analyses **FD-RBP and SD-RBP: freeze-dried and spray-dried rice bran protein SPI: soy protein isolate tein concentrate by Bera and Mukherjee (1989). leucine.4 3.2 3. Biliaderis (1983) concluded that changes in protein enthalpy could be used to predict the extent of protein denaturation.8 19.8 7. There was no significant difference in surface hydrophobicity between FD-RBP and SD-RBP (p > 0.1.37 1. FD-RBP and SD-RBP may be less denatured than similar product reported by Wang and others (1999). 84.0/10.05).99 g/100 g) in rice bran protein isolate.4 Temperature. Protein solubility The solubilities of FD-RBP. SD-RBP. Heat processing during oil extraction denatures the rice bran protein that could lead to a higher hydrophobicity values.96 J/ g for rice bran protein isolate extracted with phytase and xylanase at pH 5.05). g/100 g protein Amino acid Aspartic acid Threonine Serine Glutamic acid Glycine Alanine Valine Isoleucine Leucine Tyrosine Phenylalanine Lysine Histidine Arginine SPI** 13. and 84.0/6.9 3. SPI: soy protein isolate. *FDRBP and SD-RBP: freeze-dried and spray-dried rice bran protein.0/ 8.6 84. 10. SD-RBP. and the linear relationships between protein concentration and fluorescence intensity are shown in Figure 3.05).37 J/g.0 3.8 4.50 2.3 7. The solubilities of freeze-dried and spray-dried products were very similar from pH 2.0/4.8 5.9 4.4.7 5. amino acids in either FD-RBP or SD-RBP can meet the 2-y-old children’s requirement recommended by the FAO/WHO/UNU (Joint FAO/WHO/UNU 1985). Wang and others (1999) reported Figure 2—Effect of pH value on nitrogen solubility.05). Above pH 6.4 °C and 0.8% for SD-RBP.9 3. and Wang and others (1999).9% for FD-RBP.1 84.88 7.5 7.9 75.1 2. Data from differential scanning calorimetry (DSC) measurements for FD-RBP.0. and 1.7 3.2 3. mJ 84. and arginine. At pH 2. Wang and others (1999) measured the surface hydrophobicity of rice bran protein isolate and concentrate of full-fat nonstabilized rice bran.

Helm RM. Rice bran protein concentrates obtained by wet alkaline extraction.2b 47.Physicochemical properties and functionality of rice bran protein… Table 3—Emulsifying activities and emulsion stabilities of rice bran proteins and soy protein isolate Protein FD-RBP** SD-RBP SPI Emulsifying activity 0.5a Table 4—Foam capacity and foam stability of freeze-dried/ spray-dried dried rice bran proteins and soy protein isolate Foaming capacity. WHO. Food Chemistry and Toxicology RBP and SD-RBP. protein molecules should form continuous intermolecular polymers enveloping the air bubbles.2 kDa. 1981. Characterization of protein fractions of rice bran to devise effective methods of protein solubilization. J Food Sci 60:1066-1069. Petruccelli and Anon (1994) reported that emulsifying properties are closely associated with protein surface hydrophobicity.19b* 0. min 16. Expert Consultation. Juliano BO.4 mL) in comparison with egg white (20.6a Foaming stability. SPI: soy protein isolate. 1989. To have foam stability. 1995. Switzerland. Solubilities of both FD-RBP and SD-RBP at pH 4. Electrophoresis of rice bran protein hydrolyzate showed only 4 bands for FD-RBP and SD-RBP and their molecular weights ranged from 6. Hermansson AM. and for rice bran protein concentrate were 0. Wang and others (1999) reported that rice bran protein isolate had similar foaming properties (19.30a. Connor MA. In comparison with SPI. Geneva. JAOCS 77:779-784. and soybean oils for rice bran protein isolate from full-fat rice bran were 0. Hettiarachchy. Va.5b 17. Food Chem 10:239-265. Vol 2. Minn. and 9.10 to 4. To have foam capacity. but the foam stability (105 min) was slightly lower than that of egg white (120 min). Energy and protein requirements.20 min. Kohler GO. 1996. P 1096-1097.. It is also possible that the hydrolyzed peptides in the FD-RBP and SD-RBP products could not form cohesive layers to stabilize the foam. 8th Ed. Kato A. Method 982. Differential scanning calorimetry in food research: A review. Protein-stabilized foams and emulsions. tyrosine.0b* 4. SPI: soy protein isolate that the emulsifying activity and emulsion stability with canola. 1983. Paul. 1985. Minn. **FD-RBP and SD-RBP: freeze-dried and spray-dried rice bran protein. 2000. 1983.24a Emulsion stability. Cereal Chem 74(5):662-668. Ultrafiltration of partially hydrolyzed rice bran protein to recover value-added products. Joint FAO/WHO/UNU. Ser. Emulsifying activities and emulsion stabilities and foam capacity of FD-RBP and SD-RBP were lower than SPI. Cereal Foods World 41:839-842.3b 27. Bera MB. 15th Ed.: Association of Official Analytical Chemists. NS.05). This could be due to the lack of sufficient amounts of secondary and tertiary structures that are needed to form cohesive layers around air droplets. 1985. Agric Chem Biotechnol 42(4):180-185. . and foaming properties of rice bran protein concentrates. Burks AW. *FD-RBP and SD-RBP: freeze-dried and spray-dried rice bran protein. Rep. Foam capacities were very low for FD- pectinase and protease to produce hydrolyzate and then freeze-dried and spray-dried to obtain rice bran protein.90 to 4.37 and 3. Both FDRBP and SD-RBP had higher hydrophobicities than protein isolate and concentrate of full-fat nonstabilized rice bran but lower than SPI. since intermolecular cohesiveness and elasticity are important to produce stable foams. Association of Official Analytical Chemists [AOAC]. 1994. FD-RBP and SDRBP products could find applications as ingredients in nutraceutical products. 1978. References American Association of Cereal Chemists [AACC]. mL FD-RBP** SD-RBP SPI 4. Biliaderis CG.3 *Values are means of triplicate analyses and the values followed by different letters in the same column are significantly different (p<0. Cereal Chem 53(4):488-496.05). Kobayashi K. valine. and the foam stabilities were almost negligible. Hypoallerginicity of rice protein. J Food Sci 54(1):142-145. nd is “ not determinable ” . and foams.30 min. In: Official methods of analysis. proteins should solubilize in the aqueous phase and rapidly unfold to form a cohesive layer of protein around gas/air droplets. nr 724. Interfaces. Gnanasambandam R. emulsifying. Hamada J. WHO Tech. This could be due to the exposure of hydrophobic groups of denatured proteins. and arginine. Hamada J. This requires flexible protein molecules with few secondary and tertiary structures. Saunders RM.10 to 0. corn. Matsudomi N. St. J Food Sci 50:486-491. and SPI are given in Table 4. Paul.18b 0.min 0 0 78.51 of emulsifying activity with a stability of approximately 11 min for freeze-dried protease extracted protein hydrolysates from nonstabilized rice bran at pH 5. Solubility. Damodaran S. Conclusions H EAT-STABILIZED DEFATTED RICE BRAN WAS HYDROLYSED WITH Foam capacity and stability of proteins The foam capacities and stabilities of FD-RBP. Halling PJ. Approved methods of the American Association of Cereal Chemists. 1999. Arlington. 1997. respectively. Jang S. Relationships of hydrophobicity and net charge to the solubility of milk and soy proteins.5 and 4.30 to 0. Hamada (2000) reported 0. Method 46-08. Development of new food protein through chemical modification of rice bran proteins. both FD-RBP and SD-RBP contained higher amounts of the essential amino acids threonine. 1990.5 mL). 7. Physicochemical aspects of soy proteins structure formation.: AACC. Rice: Chemistry and technology. Mukherjee RK. 1990. Adv Food Nutr Res 34:1-79. Nakai S. J Text Stud 9:33-58. Takahashi A. P 1-2. CRC Crit Rev Food Sci Nutri 21:155-203. histidine. Bae D. Hayakawa S. SD-RBP. St. **FD-RBP and SD-RBP: freeze-dried and spray-dried rice bran protein SPI: soy protein isolate *Values are the means of triplicate analyses and values followed by different letters in the same column are significantly different (P<0.: American Association of Cereal Chemists.0 were similar but higher than SPI. Our data showed that both freeze-dried and spray-dried products had relatively good emulsifying properties and SPI had highest emulsifying activity and stability. 1976.5 to 66. 1990. protein films. Determination of foam- Figure 3—Fluorescence intensity of rice bran protein and soy protein solutions.30 to 0. Protein concentrates from nonheat-stabilized rice bran: Preparation and properties.

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