Protein Extraction from Heat-stabilized Defatted Rice Bran: II. The Role of Amylase, Celluclast, and Viscozyme

ABSTRACT: The effectiveness of 3 carbohydrases for protein extraction from heat-stabilized defatted rice bran (HDRB) was evaluated. Amylase, viscozyme and celluclast extracted a maximum of 45.4, 12.1, and 28.5% protein, respectively. Further study showed that extracted protein ranged from 9.5 to 58.4% under conditions of water to bran ratio (5:1 to 20:1), α-amylase (0 to 110000 units/10 g rice bran), temperature (35 to 55 °C), and time (1 to 8 h). The maximum protein extracted was 58.4% with a water to bran ratio of 17:1, 87637 units amylase, and 50.9 °C. These results suggest that impure food-grade amylase containing protease is more effective than celluclast and viscozyme in protein extraction from HDRB.     

Physicochemical Properties and Functionality of Rice Bran Protein Hydrolyzate Prepared from Heat-stabilized Defatted Rice Bran with the Aid of Enzymes

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.     

Amino Acid Profiles of 44 Soybean Lines and ACE‐I Inhibitory Activities of Peptide Fractions from Selected Lines

Soybeans are cultivated in the United States chiefly for cooking oil, while the residue after oil extraction (soybean meal) is mostly used in animal feed formulations. High protein content in the defatted soybean meals led to the extraction of pure protein and its application in food products. We selected 44 soybean lines to determine their moisture and protein contents, and their amino acid composition was investigated. Soybean lines with high protein content, one high yielding (R95‐1705), and two high oleic acid (N98‐4445A, S03‐543CR), were selected for protein isolate preparation, hydrolysis using alcalase and gastro‐intestinal (GI) resistance. Furthermore, the GI resistant hydrolysates were fractionated and tested for angiotensin‐I‐converting enzyme (ACE‐I) inhibition activity. The amino acid analysis showed high methionine in the high protein and fatty acid lines (R05‐4494 and R05‐5491), and high cysteine content in one of the high oleic acid soybean line CRR05‐188 in comparison to the check lines (UA‐4805 and 5601‐T). The protein isolate with the highest purity (90–93 %) was derived from the selected lines N98‐4445A and S03‐543CR, and hydrolyzed using alcalase enzyme. The protein hydrolysates (500 µg/mL) showed inhibition of the ACE‐I by 49 %. The results from this study will promote the use of high oleic acid soybeans as a source of protein and peptides with functional activities.     

Dietary fiber supplements: Effects on serum and liver lipids and on liver phospholipid composition in rats

Rats (6 per group) were fed semipurified diets containing either particulate fibers (alfalfa, 10%; cellulose, 10%; bran, 10%), a soluble ionic fiber (pectin 5%), soluble, nonionic fibers (guar gum, 5%; Metamucil, 10%), a mixed fiber preparation (Fibyrax, 10%, or an insoluble, ionic bile acid-binding resin (cholestyramine, 2%). The control group was fed the unsupplemented diet. The feeding period, during which diet and water were provided ad libitum, was 28 days. Compared with the control group, serum total cholesterol levels were increased by more than 10% in rats fed alfalfa and decreased by more than 10% in rats fed cellulose, guar gum, Fibyrax and cholestyramine. There were no significant differences in percentage of plasma HDL cholesterol. Serum triglycerides were elevated in the groups fed alfalfa, pectin, guar gum or Fibyrax and reduced in the group fed Metamucil. Plasma phospholipids were elevated in rats fed alfalfa or bran, unaffected in rats fed pectin or Metamucil and reduced in the other groups. Liver total cholesterol was elevated in all groups but those fed wheat bran and cholestyramine. The percentage of liver cholesterol present as ester was elevated in every group except that fed cholestyramine. Liver triglycerides were reduced in rats fed guar gum or Metamucil and elevated in those fed alfalfa. Liver phospho-lipids were lowered in the group fed cellulose. Liver phospholipids were fractionated by thin layer chromatography to give phosphatidylcholine (PC), phosphatidylethanolamine (PE), sphingomyelin (Sph), lysophosphatidylcholine (LPC) and phosphatidylinositol plus phosphatidylserine (PI+PS). PC was elevated in all test groups (7–25%); PE levels ranged from 14% below to 0.3% above controls; Sph levels were sharply lower (20–53%) in all groups. LPC and PI+PS levels were close to the control value in all test groups. The results demonstrate that different dietary fibers can affect liver phospholipid composition. In view of the critical roles of phospholipids in many biological reactions, it will be interesting to survey the influence of dietary fiber on phospholipid spectra of other tissues. Deceased.     

Recovery of fish oil-derived fatty acids in lymph of thoracic duct-cannulated wistar rats

The absorption of equivalent doses of eicosapentaenoic and docosahexaenoic acids was compared in rats when administered as the ethyl ester concentrate, ethyl ester concentrate plus olive oil, free fatty acid or triacylglycerol (menhaden oil). Lymph was collected from a thoracic duct cannula for 24 hr after dosing via an indwelling duodenal catheter. After 24 hr, the absorption of eicosapentaenoic acid was greater for the free fatty acid and menhaden oil than for the ethyl ester form, but docosahexaenoic acid absorption was comparable for all forms. Other rats had greater plasma levels of eicosapentaenoic and docosahexaenoic acids 5 hr after oral gavage dosing with menhaden oil than did rats dosed with the ethyl ester form.     

Effect of peanut oil and randomized peanut oil on cholesterol and oleic acid absorption, transport, and distribution in the lymph of the rat

Peanut oil was shown to be atherogenic in cholesterol-fed rats, rabbits, and monkeys. However, after randomization, a process in which the fatty acids in peanut oil are randomly rearranged, its atherogenicity was significantly reduced in cholesterol-fed rabbits and monkeys. The mechanism for this effect remains unknown. This study was designed to investigate whether the absorption, transport and distribution of dietary cholesterol and oleic acid in the lymph were altered in the presence of peanut oil or randomized peanut oil. Previous investigators collected lymph through the mesenteric duct for 6 h and analyzed lymph for cholesterol. In the present study, lymph fluids were collected at timed intervals for up to 8 h and then at 24 h via the thoracic duct. Cholesterol and oleic acid (fatty acid) were estimated not only in the whole lymph but also in lymph lipoprotein fractions and in major lipid fractions. A 24-h lymph collection will enhance accuracy as short-term fluctuations in lipid absorption will not affect the results. Thoracic duct lymph collection is quantitative compared to mesenteric duct lymph collection, which provides only a fraction of the total lymph. Rats were given a lipid emulsion containing either peanut oil or randomized peanut oil. The emulsion also contained cholesterol, oleic acid, and sodium taurocholate in saline and was given through a duodenal catheter. Results show that absorption, transport, and distribution of cholesterol and oleic acid in the lymph fluids were similar in both dietary groups. These results suggest that the atherogenicity of peanut oil may be due to other events taking place subsequent to the release of cholesterol-containing chylomicrons and very low density lipoprotein by the small intestinal epithelial cells into the blood or may be due to the triglyceride structure itself.     

Comparison of the clearances of serum chylomicron triglycerides enriched with eicosapentaenoic acid or oleic acid

Rat mesenteric lymph chylomicrons containing triglycerides enriched with either [¹⁴C]oleic acid (OA) or [¹⁴C]-eicosapentaenoic acid (EPA) were prepared by ultracentrifugation of lymph samples collected for 6 hr after a single duodenal infusion of an emulsion containing either fatty acid. These chylomicrons were injected into the jugular vein of recipient rats and, at various time intervals, blood was drawn and serum was assayed for radioactivity. In separate animals, serum lipoprotein fractions were separated by ultracentrifugation, and the redistribution of labeled fatty acid among circulating lipoproteins was determined by liquid scintillation spectrometry. When the early disappearance rates (10 min) of either total serum radioactivity or specifically the chylomicron fraction were compared, there were no differences between the groups receiving OA-or EPA-enriched chylomicrons. However, disappearance rates of EPA-enriched chylomicrons were slower than those of OA-enriched chylomicrons from 25 to 90 min. The small but significant differences in the disappearance rates for the longer time periods cannot be ascertained without further studies. At 5 min after injection of either type of chylomicron, the d<1.006 g/ml lipoprotein fraction of serum chylomicrons and very low density lipoproteins contained almost 90% of the original radioactivity. By 240 min, when less than 2% of the radioactivity remained, this radioactivity in the d<1.006 g/ml fraction was 43–46%, with concomitant increases in the low and high density lipoprotein fractions and in the lipoprotein-free serum. Deceased.