Comparison of interlaboratory variation in amino acid analysis and rat growth assays for evaluating protein quality

Estimates of inter- and intralaboratory variation of protein efficiency ratio (PER), relative PER (RPER), net protein ratio (NPR), relative NPR (RNPR), and nitrogen utilization (NU) were compared with those of amino acid analysis in the same batches of 7 protein sources (ANRC casein, egg white solids, minced beef, soy assay protein, rapeseed protein concentrate, pea flour, and whole wheat flour). Interlaboratory variation (estimated as between-laboratories coefficients of variation, CV) of NPR and RNPR (up to 6.0%) was lower than that of PER (up to 20.2%) and RPER (up to 18.5%). The interlaboratory determination of NPR and RNPR was also more reproducible than that of most essential amino acids (CV up to 10.0%), especially tryptophan (CV up to 23.7%), cystine (CV up to 17.6%), and methionine (CV up to 16.1%). Intralaboratory variation (estimated as within-laboratories CV) of amino acid analysis (up to 4.7%), however, was comparable to that of protein quality indices in most protein sources (up to 6.0%). The significant (P less than 0.01) positive correlations (r = 0.68-0.74) between amino acid scores and protein quality indices based on rat growth were further improved when amino acid scores were corrected for digestibility of protein (r = 0.73-0.78) or individual amino acids (r = 0.79-0.82).     

大豆蛋白限制性酶解模式与产品胶凝性的相关性

为了改善大豆蛋白的胶凝性,对大豆浓缩蛋白、大豆分离蛋白进行限制性酶解处理,并考察相应产品的蛋白酶酶解模式与胶凝性变化的相关性。该研究以蛋白质的水解度为指标,通过中性蛋白酶、胰蛋白酶的酶解作用,水解大豆浓缩蛋白、大豆分离蛋白至蛋白质水解度(DH)为1%、2%,考察酶性质、蛋白质的DH对产品胶凝性影响,并利用SDS-凝胶电泳进行确认。结果表明:大豆浓缩蛋白经中性蛋白酶、胰蛋白酶的酶解后,产品胶凝性均显著下降;大豆分离蛋白经中性蛋白酶的酶解后,产品胶凝性在DH为1%时增加,但在DH为2%时下降;大豆分离蛋白经胰蛋白酶酶解后,胶凝性显著改善。SDS-凝胶电泳确认,蛋白质的酶水解模式和水解度不同是导致产品胶凝性产生不同变化的原因。     

Amino acid rating method for evaluating protein adequacy of infant formulas

Amino acid profiles and/or protein digestibility (by the rat balance method) were determined for various forms (powder, ready-to-use, liquid concentrate, etc.) of cow's milk- and soy-based infant formulas obtained from 4 manufacturers. The essential amino acid data of the formulas were compared with that of human milk for the calculation of amino acid scores (based on the single most limiting amino acid). The product of amino acid score and total protein (g/100 kcal) was then termed "amino acid rating." Amino acid scores for the milk- and soy-based formulas ranged from 59 to 90 and from 59 to 81%, respectively, due to deficiencies in sulfur amino acids and/or tryptophan. Because of significantly higher total protein contents (g/100 kcal) of soy- (2.65-3.68) and milk-based (2.20-2.95) infant formulas compared to human milk (1.5), the relative amino acid ratings (human milk = 100) for all infant formulas except 2 liquid concentrates (having values of 87%) were above 100%. Values for true digestibility of protein in milk- and soy-based formulas ranged from 87 to 97 and from 92 to 95%, respectively. When corrected for protein digestibility, the relative amino acid ratings for all the milk-based liquid concentrates were below 100% (77-98%).     

Chemical,Physicochemical, and Nutritional Evaluation of Plantago (Plantago ovata Forsk)

Plantago ovata F. are small tan‐colored seeds with ≈30% weight husk. Plantago's husk high content of soluble fiber makes it a good lubricant of the intestinal track with demonstrated effects in lowering plasma cholesterol levels in humans and experimental animals. Plantago seeds grown in Northern Mexico were analyzed for proximate composition, combustion heat, soluble and insoluble dietary fiber, fatty acids, amino acids, and protein fractionation. In vitro digestibility and digestibility of dry matter, apparent and true digestibility, and net protein ratio (in vivo) were also analyzed. Plantago seeds had 17.4% protein, 6.7% fat, 24.6% total dietary fiber, 19.6% insoluble fiber, 5.0% soluble fiber, and a combustion heat of 4.75 kcal/g. Osborne fractionation (based on solubility) yielded albumin 35.8%, globulin 23.9%, and prolamin 11.7%. The oil from plantago seeds had a high percentage of linoleic acid (40.6%) and oleic acid (39.1%) and a minor proportion of linolenic acid (6.9%). In vitro protein digestibility of the plantago seed was 77.5%, suggesting a highly digestible protein. Lysine content was 6.82 g/100 g of protein, higher than wheat and oats (2.46 and 4.20 g/100 g of protein, respectively). Rat bioassays showed values of 89.6% digestibility of dry matter, 86.0% apparent digestibility, 88.1% true digestibility, and 4.40 net protein ratio corrected (NPRc). The importance of these findings is that plantago whole grain shows favorable nutritional quality when compared with cereals and legumes.     

Yeast (Saccharomyces cerevisiae) protein concentrate: preparation, chemical composition, and nutritional and functional properties

The main objective of the present study was to compare the composition and functional and nutritional properties of whole yeast cells (WY) from an ethanol distillery with those of a phosphorylated protein concentrate (PPC) prepared from the same cells. Comparisons were also made of PPC with texturized soy protein (TSP) and soy protein isolate (SPI), both acquired in the local market. Yeast (Saccharomyces cerevisiae) is a rich source of protein, soluble fiber, and some minerals. Saturated fatty acids predominated over monounsaturated and polyunsaturated in both WY and PPC. The functional properties of PPC were similar to those of SPI and TSP. Both soy products and PPC replaced 20 or 40% chuck roll protein without affecting the emulsion properties of the meat products. Amino acid scoring was high for both WY and PPC; digestibility was higher (90%) for PPC and lower (68%) for WY. The protein nutritive value of PPC did not differ from that of casein and was significantly higher than that for WY.     

Preparation and functional properties of rice bran protein isolate

Rice bran protein isolate (RBPI) containing approximately 92.0% protein was prepared from unstabilized and defatted rice bran using phytase and xylanase. The yield of RBPI increased from 34% to 74.6% through the use of the enzymatic treatment. Nitrogen solubilities of RBPI were 53, 8, 62, 78, 82, and 80% at pHs 2.0, 4.0, 6.0, 8.0, 10.0, and 12.0, respectively. Differential scanning calorimetry showed that RBPI had denaturation temperature of 83.4 degrees C with low endotherm (0.96 J/g of protein). RBPI had similar foaming properties in comparison to egg white. But emulsifying properties of RBPI were significantly lower than those of bovine serum albumin. The result of amino acid analysis showed that RBPI had a similar profile of essential amino acid requirements for 2-5-year-old children in comparison to that of casein and soy protein isolate.     

青鳞鱼骨粉的食用营养价值及应用研究

研究了青鳞鱼骨粉的食用营养价值及其在玉米粥、马蹄粉和鸡精中的应用。结果表明：(1)青鳞鱼骨粉是一种蛋白质含量为27.0%、脂肪含量为2.0%的典型的高营养低热能食品；它具有独特的补钙生物学功能的关键因素和优势。(2)青鳞鱼鱼骨粉钙磷比为2.32∶1，钙和铁易于被人体吸收和利用。(3)青鳞鱼骨粉氨基酸总量为24.36 mg/100g，其中必需氨基酸为5.23 mg/100g，占氨基酸总量的21.49%；蛋白质质量优越于猪骨粉，具有更大的利用价值。(4)以10%的比例将超微鱼骨粉分别添加到策友牌玉米粥，互润牌桂花马蹄粉和美极牌鲜鸡精中，可以提高产品的感官质量，达到强化蛋白质和钙磷铁等矿物质元素、改善食品的钙磷比例等目的。     

Metabolism of theanine, gamma-glutamylethylamide, in rats

The metabolism of theanine, one of the major amino acid components in tea (Camellia sinensis), was studied in rats. High-performance liquid chromatography (HPLC) with fluorometric detection was used to evaluate the nature of theanine's metabolites in plasma, urine, and tissues. In the urine samples collected after administration of 100, 200, and 400 mg each of theanine, intact theanine, L-glutamic acid, and ethylamine, these compounds were detected in a dose-dependent manner. When 200 mg of theanine was orally administered to rats, the plasma concentrations of theanine and ethylamine reached their highest levels about 0.5 and 2 h after administration, respectively. It seems most likely that the enzymatic hydrolysis of theanine to glutamic acid and ethylamine was accomplished in the kidney. These results indicate that orally administered theanine is absorbed through the intestinal tract and hydrolyzed to glutamic acid and ethylamine in the rat kidney.     

Inhibition of iron uptake by phytic acid, tannic acid, and ZnCl2: studies using an in vitro digestion/Caco-2 cell model.

The objective of this study was to document the effects of phytic acid, tannic acid, and zinc on iron uptake in an in vitro digestion/Caco-2 cell culture model. The effects of phytic acid and tannic acid on iron uptake were measured at increasing molar ratios of FeCl3 to phytic acid or tannic acid. Maximal inhibition of iron uptake by phytic acid occurred at a 1:10 ratio of Fe to phytic acid. Dialyzable Fe decreased with a low Fe to phytic acid ratio but increased with Fe:phytic acid ratios greater than 1:3 indicating that more iron was soluble at higher phytic acid levels but less available. As in human studies, heme iron was less inhibited by phytic acid than nonheme iron. Tannic acid was a more potent inhibitor of nonheme iron uptake, as maximal inhibition (97.5%) of iron uptake occurred at a ratio of 1:1 or less. The addition of ZnCl2 to the digest at ratios of 1:0.5 and 1:1 decreased iron uptake by 57 and 80%, respectively. Overall, the results agree qualitatively with studies in humans and demonstrate the relative effects of these compounds on iron uptake in this model system. This study provides key information for determining iron availability under more complex meal conditions.     

聚合氨基酸对北方水稻土中氧化铁存在形态的影响

为了解不同类型水稻土中氧化铁含量特征,探明添加外源聚合氨基酸对水稻土中氧化铁形态变化的影响,本研究以中国北方不同类型水稻土(棕壤型、草甸土型和滨海盐渍型)为研究对象,设置添加占供试土壤干重0.05%的γ-聚谷氨酸和聚天冬氨酸处理,以不添加氨基酸作为对照。通过室内恒温厌氧培养30 d后,测定各水稻土全铁、游离氧化铁、无定形氧化铁和络合态铁含量及氧化铁的活化和络合程度。结果表明:供试的北方3种典型水稻土中,游离氧化铁含量为滨海盐渍型草甸土型≥棕壤型,络合态铁含量为棕壤型滨海盐渍型≥草甸土型,而无定形氧化铁含量为棕壤型滨海盐渍型草甸土型。两种外源聚合氨基酸对不同类型水稻土氧化铁形态转化能力影响存在差异,与不添加氨基酸的对照相比,添加γ-聚谷氨酸的棕壤型水稻土无定形氧化铁和络合态铁含量分别增加27.72%和32.25%,聚天冬氨酸对无定形氧化铁和络合态铁含量无显著促进作用;在草甸土型水稻土中,γ-聚谷氨酸和聚天冬氨酸均能显著增加无定形氧化铁含量,且与对照相比,络合态铁含量分别增加136.24%和12.00%;γ-聚谷氨酸能有效促进滨海盐渍型水稻土中无定形氧化铁和络合态铁的生成。总之,添加γ-聚谷氨酸和聚天冬氨酸对水稻土游离氧化铁的含量没有明显影响;而添加γ-聚谷氨酸能有效增加水稻土中无定形氧化铁和络合态铁含量,降低晶胶率,有利于提高土壤中有效铁含量,显著活化铁氧化物,抑制各类型水稻土中铁的结晶老化;而聚天冬氨酸对水稻土无定形氧化铁和络合态铁没有明显的激发效应。     

复合氨基酸微量元素螯合肥制备工艺研究

以废弃鸡毛水解液为复合氨基酸来源,研究其与微量元素铁、锌、锰、铜螯合的最佳工艺条件,并利用有机溶剂沉淀分离法制备固体复合氨基酸亚铁、锌、锰、铜螯合物,以螯合率为指标考察影响制备工艺的主要因素。结果表明,最佳工艺参数为:复合氨基酸与亚铁、锌、锰和铜螯合反应的最佳配位体摩尔比均为2∶1;最佳pH值范围分别为5.0、8.0、4.0～5.0和6.0～7.0;反应温度为室温(222～6℃);反应时间为30.min。在上述条件下,复合氨基酸与各微量元素的螯合率均超过95%;所得复合氨基酸螯合物可与0.2.mol/L磷酸二氢钾复配,克服了传统微肥完全不能与磷酸盐复配的缺陷。     

Determination of sialic acid by the thiobarbituric acid reaction in sweet whey and its fractions.

Determination of sialic acid in sweet whey is useful as the concentration of sialic acid reflects the amount of glycomacropeptide (GMP) present. In this study, the concentration of total sialic acid was determined by the thiobarbituric acid reaction after dialysis of samples in water, and the concentration of GMP sialic acid was estimated by gel chromatography on Sephacryl S-200. Concentrations of total and GMP sialic acid determined in a sweet whey sample prepared from fresh milk were 2.0 and 1.5 microg/mg of dry weight, respectively. Analysis of commercial samples showed that the concentration of total sialic acid in sweet whey was 9 times lower than that in whey protein concentrate but 18 times higher than that in whey permeate. A similar trend was observed in the variation of GMP sialic acid concentration between sweet whey and why protein concentrate. The concentration of sialic acid differed 10 times between two samples of whey protein isolate.     

Isoflavone-free soy protein prepared by column chromatography reduces plasma cholesterol in rats

To know whether isoflavones are responsible for the hypocholesterolemic effect of soy protein, the effect on plasma cholesterol of isoflavone-free soy protein prepared by column chromatography was examined in rats. Five-week-old male Sprague-Dawley rats were fed cholesterol-enriched AIN-93G diets containing either 20% casein (CAS), 20% soy protein isolate (SPI), 20% isoflavone-free SPI (IF-SPI), 19.7% IF-SPI + 0.3% isoflavone-rich fraction (isoflavone concentrate, IC), or 20% CAS + 0.3% IC for 2 weeks. Plasma total cholesterol concentrations of rats fed SPI and IF-SPI were comparable and were significantly lower than that of rats fed CAS. The addition of IC to the CAS and IF-SPI did not influence plasma cholesterol level. Fecal steroid excretion of the three SPI groups was higher than that of the two CAS groups, whereas the addition of IC showed no effect. Thus, a significant fraction of the cholesterol-lowering effect of SPI in rats can be attributed to the protein content, but the isoflavones and other minor constituents may also play a role.     

Absorption of chlorogenic acid and caffeic acid in rats after oral administration

Absorption of orally administered chlorogenic acid (5-caffeoylquinic acid) and caffeic acid in rats was studied to obtain plasma pharmacokinetic profiles of their metabolites. Rats were administered 700 micromol/kg body weight of chlorogenic or caffeic acid, and blood was collected from the tail for 6 h after administration. Ingested caffeic acid was absorbed from the alimentary tract and was present in the rat blood circulation in the form of various metabolites. On the other hand, only traces of metabolites, supposedly caffeic and ferulic acids conjugates, were detected in rat plasma for 6 h after chlorogenic acid administration. Chlorogenic acid and small amounts of caffeic acid were found in the small intestine for 6 h after chlorogenic acid administration. These results suggest that chlorogenic acid is not well absorbed from the digestive tract, unlike caffeic acid, and subject to almost no structural changes to the easily absorbed forms.     

A possible mechanism for humic acid action on cell elongation in root segments of Pisum sativum under aseptic conditions

At low concentrations (up to 50 mg/l) humic acid enhances cell elongation in excised pea root segments, but at higher concentrations it is inhibitory. At concentrations stimulating cell elongation humic acid has no effect on protein metabolism but does inhibit the formation of cell-wall bound hydroxyproline from proline. Humic acid does not stimulate cell elongation or inhibit the formation of cell-wall bound hydroxyproline in the presence of ferrous iron. Results similar to those of humic acid were obtained using the iron chelator αα'-dipyridyl, but γγ'-dipyridyl. which does not chelate iron. was without effect on cell elongation.The cessation of cell elongation corresponds to the time at which there is a substantial increase in cellwall bound hydroxyproline, this latter process requiring ferrous iron. Thus humic acid may enhance growth by virtue of its ability to complex ferrous iron within the plant tissues.     

Properties of protein powders from arrowtooth flounder (Atheresthes stomias) and herring (Clupea harengus) byproducts

Functional, nutritional, and thermal properties of freeze-dried protein powders (FPP) from whole herring (WHP), herring body (HBP), herring head (HHP), herring gonad (HGP), and arrowtooth flounder fillets (AFP) were evaluated. The FPP samples have desirable nutritional and functional properties and contained 63-81.4% protein. All FPP samples had desirable essential amino acid profiles and mineral contents. The emulsifying and fat adsorption capacities of all FPP samples were higher than those of soy protein concentrate. The emulsifying stability of WHP was lower than that of egg albumin but greater than that of soy protein concentrate. Thermal stability of the FPP samples is in the following order: HGP > HBP > WHP > HHP > AFP.     

Chlorogenic acid moderately decreases the quality of whey proteins in rats

During processing and storage, phenolic compounds (PCs) may react with food protein bound amino acids (AAs). Such reactions have been reported to change physicochemical and to decrease in vitro digestion properties of proteins. A rat growth and nitrogen (N) balance study was conducted to prove whether derivatization with chlorogenic acid (CA) affects the nutritional quality of beta-lactoglobulin (beta-LG). Test diets (10% protein level) contained nonderivatized beta-LG (LG, treated under omission of CA), low derivatization level beta-LG (LGL), high derivatization level beta-LG (LGH), or casein supplemented with l-methionine (0.3% of diet; C+met) as an internal standard. An additional group received untreated beta-LG supplemented with pure CA (1.03% of diet; LG+CA). The AA composition of test proteins, plasma AAs, and liver glutathione (GSH) concentrations were determined. Protein digestibility-corrected amino acid score (PDCAAS) was calculated using human or rat AA requirement patterns and rat fecal digestibility values. N excretion was significantly higher in feces and lower in urine of rats fed with LGH as compared to LG and LGL. Consequently, true N digestibility (TND) was significantly lower with LGH as compared to LG and LGL. The lower content of methionine, cysteine, lysine, and tryptophan in LGH corresponded to a reduced TND. Net protein utilization (NPU) was not different between treated beta-LG fed diet groups but was lower than in LG+CA and C+met fed groups. Only at a relatively high level of derivatization with CA, the otherwise good nutritional quality of beta-LG is affected so that TND is reduced, while NPU still remains unaffected. Derivatization of beta-LG with CA does not seem to lead to an additional deficiency in a specific indispensable AA in growing rats fed with 10% protein.     

Behavior of protein in the presence of calcium during heating of whey protein concentrate solutions

The effect of added CaCl(2) on heat-induced changes in whey protein (WP) solutions prepared from whey protein isolate (WP1), acid whey protein concentrate (WP2), and cheese whey protein concentrate (WP3) was investigated. The loss of native-like, proteins, aggregation, and gel firmness of WP were maximum at certain levels of added CaCl(2). These levels were different for different WP products. The effect of added CaCl(2) on these changes appeared to be related to the initial calcium concentrations of these solutions. The higher the calcium content of the product, the less available sites for added CaCl(2) to bind. It was considered that addition of CaCl(2) changed the types of protein interactions that formed the protein aggregates during heating. Added calcium caused dramatic decreases in fracture stress of WP gels due to the formation of large protein aggregates.     

有机废液对镉污染土壤上白菜生长和镉积累影响

为探究有机废液在镉污染土壤施用对作物生长和环境健康风险的影响，该研究针对豌豆发酵液、玉米浆浓缩液、鸡粪浓缩沼液、糖蜜液和氨基酸母液5种价格低廉且在农业中应用广泛的有机废液，通过盆栽试验分析了施用2 ml/kg的5种有机废液对白菜生长与镉吸收的影响，并从白菜光合作用及抗氧化酶活性等角度进行了效果对比研究。结果表明，除氨基酸母液处理外，豌豆发酵液、玉米浆浓缩液、鸡粪浓缩沼液、糖蜜液处理较对照均可显著增加镉胁迫下白菜叶片净光合速率（P<0.05），促进白菜生长，其中施用玉米浆浓缩液分别使白菜地上部和地下部干质量提高了21.66%和68.75%。与对照相比，施用玉米浆浓缩液使白菜地上部的镉浓度降低了22.06%，并显著提高了白菜超氧化物歧化酶和过氧化氢酶活性（P<0.05），糖蜜液和氨基酸母液处理没有降低白菜镉浓度。施用玉米浆浓缩液、鸡粪浓缩沼液、糖蜜液和氨基酸母液均可显著降低白菜丙二醛含量（P<0.05），提高土壤有效态镉含量。综上，5种有机废液对镉污染条件下白菜生长和镉吸收转运的影响不同，玉米浆浓缩液促进白菜生长并减少镉积累效果最好，氨基酸母液效果最差。     

Effect of additives on subcritical water hydrolysis of whey protein isolate

The objective was to examine the effect of the additives acetic acid, lactic acid, sodium bicarbonate, sodium chloride, and sodium hydroxide on the hydrolysis of whey protein isolate with subcritical water. A screening experimental design was used to study the effect of temperature, time, and additives. The most influential additive, sodium bicarbonate, along with temperature and time was used in a second experimental design to predict the treatment conditions to maximize the degree of hydrolysis and production of free amino acids. The maximum degree of hydrolysis achieved was 50% at a concentration of 1.24 M sodium bicarbonate, 291 °C, and 28 min. The highest concentration of total amino acids was 83.0 mg/g of whey protein isolate with 0.83 M sodium bicarbonate at 264 °C for 29 min. Compared to water alone, sodium bicarbonate increased the degree of hydrolysis 4-fold and the production of amino acids by 44% and decreased peptides' molecular weight.