Alteration of kafirin and kafirin film structure by heating with microwave energy and tannin complexation

Heating with microwave energy and tannin complexation of kafirin both increase the tensile strength of cast kafirin bioplastic films. The effects of these treatments on the molecular structure of kafirin and of kafirin in the film were investigated. SDS-PAGE of heated wet kafirin showed an increase in kafirin oligomers. Disulfide groups increased in heated kafirin and in films made from the heated kafirin. Fourier transform infrared (FTIR) spectroscopy of heated kafirin and films made from the heated kafirin indicated an increase in beta-sheet conformation. In contrast, kafirin complexation with tannic acid (TA) and sorghum condensed tannin (SCT) resulted in a slight decrease in beta-sheet conformation in the kafirin and a larger decrease in the kafirin in the films. Raman spectroscopy showed that, with TA, there was a shift in peak from 1710 to 1728 cm(-1) for kafirin-tannic acid complexes, indicating kafirin and tannic acid interaction. The protein conformational changes presumably facilitated cross-linking between kafirin molecules and/or between kafirin and the tannins. Thus, although both heating with microwave energy and tannin complexation cause cross-linking of kafirin to increase film tensile strength, their effects on kafirin structure appear to be different.     

Sequential in vitro pepsin digestion of uncooked and cooked sorghum and maize samples

An in vitro protein digestion study, using pepsin, was carried out in uncooked and cooked sorghum and maize flour samples. The digestibility values from the uncooked samples showed that sorghum presents digestibility values similar to those of maize. In the case of the cooked samples, it was found that a wet cooking procedure promotes a decrease in sorghum protein digestibility when compared to maize. Electrophoresis was used to follow the in vitro pepsin sequential digestion procedure, and infrared spectroscopy was applied to establish its efficiency. SDS-PAGE results showed that both uncooked samples (sorghum and maize) behave in a similar way. The wet cooking procedure increases the amount of high molecular weight aggregates and promotes the appearance of two nonreducible and nondigestible 45 and 47 kDa proteins. These two protein fractions are directly related to the loss of digestibility. It was also shown that in cooked sorghum the monomers (gamma-, alpha-, and beta-) are more resistant to digestion than the corresponding uncooked samples.     

Effect of preparation conditions on protein secondary structure and biofilm formation of kafirin

Various extraction and drying conditions for the isolation of kafirin from dry-milled, whole grain sorghum have been investigated, with a view to optimizing extraction of the protein for commercial food coatings and packaging films. The addition of sodium hydroxide to an aqueous ethanol extractant increased the yield and solubility of kafirin. Subsequent heat drying at 40 degrees C was shown to cause the kafirin to aggregate as indicated by an increase in intermolecular beta-sheets. Extraction of the flour using ethanol (70%, w/w) with 0.5% (w/w) sodium metabisulfite and 0.35% (w/w) sodium hydroxide at 70 degrees C followed by freeze-drying of the protein was found to produce a yield of 54% kafirin with good film-forming properties. The kafirin films were assessed for their sensory properties, tensile strength, strain, and water vapor permeability. Fourier transform infrared spectroscopy was used to study the secondary structure of the extracted kafirins. The best films were made with kafirin containing a large proportion of nativelike alpha-helical structures with little intermolecular beta-sheet content as indicated by the Fourier transform infrared reflectance peak intensity ratios associated with these secondary structures. The principal factor affecting the secondary structure of the protein appeared to be the temperature at which the protein was dried. Heat drying resulted in a greater proportion of intermolecular beta-sheets. Any industrial-scale extraction must therefore minimize protein aggregation and maximize native alpha-helical structures to achieve optimal film quality.     

Improvement in water stability and other related functional properties of thin cast kafirin protein films

Improvement in the water stability and other related functional properties of thin (<50 μm) kafirin protein films was investigated. Thin conventional kafirin films and kafirin microparticle films were prepared by casting in acetic acid solution. Thin kafirin films cast from microparticles were more stable in water than conventional cast kafirin films. Treatment of kafirin microparticles with heat and transglutaminase resulted in slightly thicker films with reduced tensile strength. In contrast, glutaraldehyde treatment resulted in up to a 43% increase in film tensile strength. The films prepared from microparticles treated with glutaraldehyde were quite stable in ambient temperature water, despite the loss of plasticizer. This was probably due to the formation of covalent cross-linking between free amino groups of the kafirin polypeptides and carbonyl groups of the aldehyde. Thus, such thin glutaraldehyde-treated kafirin microparticle films appear to have good potential for use as biomaterials in aqueous applications.     

Release of protein, lipid, and vitamin E from almond seeds during digestion

The evaluation of the bioaccessibility of almond nutrients is incomplete. However, it may have implications for the prevention and management of obesity and cardiovascular disease. This study quantified the release of lipid, protein, and vitamin E from almonds during digestion and determined the role played by cell walls in the bioaccessibility of intracellular nutrients. Natural almonds (NA), blanched almonds (BA), finely ground almonds (FG), and defatted finely ground almonds (DG) were digested in vitro under simulated gastric and gastric followed by duodenal conditions. FG were the most digestible with 39, 45, and 44% of lipid, vitamin E, and protein released after duodenal digestion, respectively. Consistent with longer residence time in the gut, preliminary in vivo studies showed higher percentages of nutrient release, and microscopic examination of digested almond tissue demonstrated cell wall swelling. Bioaccessibility is improved by increased residence time in the gut and is regulated by almond cell walls.     

Transgenic expression of bean alpha-amylase inhibitor in peas results in altered structure and immunogenicity

The development of modern gene technologies allows for the expression of recombinant proteins in non-native hosts. Diversity in translational and post-translational modification pathways between species could potentially lead to discrete changes in the molecular architecture of the expressed protein and subsequent cellular function and antigenicity. Here, we show that transgenic expression of a plant protein (alpha-amylase inhibitor-1 from the common bean (Phaseolus vulgaris L. cv. Tendergreen)) in a non-native host (transgenic pea (Pisum sativum L.)) led to the synthesis of a structurally modified form of this inhibitor. Employing models of inflammation, we demonstrated in mice that consumption of the modified alphaAI and not the native form predisposed to antigen-specific CD4+ Th2-type inflammation. Furthermore, consumption of the modified alphaAI concurrently with other heterogeneous proteins promoted immunological cross priming, which then elicited specific immunoreactivity of these proteins. Thus, transgenic expression of non-native proteins in plants may lead to the synthesis of structural variants possessing altered immunogenicity.     

Investigation and optimization of the factors influencing sorghum protein extraction

To optimize the extraction of sorghum proteins, several variables were examined: sample-to-solvent ratio, detergent type and concentration, reducing agent type and concentration, extraction time, and buffer pH and concentration. Samples were quantified and characterized by RP-HPLC, FZCE, and nitrogen analysis. These studies revealed that pH, detergent type, reducing agent type, and sample-to-solvent ratio all had significant effects on the levels of protein extracted. Increasing SDS concentration (2%) and solvent-to-flour ratio (20:1) with multiple 5 min extracts reduced extraction time by 35-80% while still extracting the same levels of total protein relative to the control methodology. Reproducibility using the multiple extractions was found to be excellent with relative standard deviations of <2% for consecutive extractions.     

甜高粱茎秆残渣生料多菌种固态发酵生产蛋白饲料

为实现甜高粱茎秆残渣生产蛋白饲料的规模化应用,该研究将黑曲霉(Aspergillus niger)、里氏木霉(Trichoderma reesei),产朊假丝酵母(Candida utilis)和干酪乳杆菌(Lactobacillus casei)进行优化组合,添加不同质量分数尿素对甜高粱茎秆残渣进行生料固态发酵生产蛋白饲料。试验通过对比发酵前后粗蛋白、真蛋白、粗灰分和粗脂肪质量分数变化发现:添加4种菌的复合菌株,再添加1%尿素发酵8 d后能使以甜高粱茎秆残渣为底物的饲料中纤维素由33.00%降低至24.09%,半纤维素由20.99%降低至17.69%;粗蛋白质量分数由2.27%提升至7.14%,真蛋白由2.01%提升至6.41%。该研究简化了甜高粱秸秆残渣规模化生产饲料蛋白的工艺,为该工艺的推广应用奠定基础。     

Arsenic-contaminated soils phytotoxicity studies with sunflower and sorghum

Background, Aim and Scope  Environmental pollution caused by arsenic (As) is a major ecological problem. There has been intense worldwide effort to find As-hyperaccumulating plants that can be used in phytoremediation—the green-plant-assisted removal of chemical pollutants from soils. For phytoremediation, it is natural to prefer cultivated rather than wild plants, because their agriculture is well known. This study was conducted to evaluate the tolerance of common sunflower(Helianthus annuus L.) and sugar sorghum(Sorghum saccharatum Pers.) for soil-As contents of 10–100 mg As kg^-1 soil, with sodium arsenite as a model contaminant. Methods  Plants were grown in a growth chamber for 30 days. Microfield experiments were conducted on experimental plots. To study the phytoremediation effect of the auxins indole-3-acetic acid (IAA) and 2,4-dichlorophenoxyacetic acid (2,4-D), we treated 1- and 3-day-old plant seedlings with water solutions of the auxins (concentrations 10^-5, 10^-7, and 10^-9 g l^-1). The soil and plant-biomass samples were analyzed for total As by using the color reaction of ammonium molybdate with As. Results and Discussion  Phytotoxicity studies showed that 100 mg As kg^-1 soil poisoned sunflower and sorghum growth by 50%. There was a linear correlation between soil-As content and As accumulation in the plants. Laboratory experiments showed that the soil-As content was reduced two- to threefold after sunflower had been grown with 10–100 mg As kg^-1 soil for 30 days. Treatment of sunflower and sorghum seedlings with IAA and 2,4-D at a concentration of 10^-5 g l^-1 in microfield experiments enhanced the phytoremediation two- to fivefold as compared with untreated control plants. The best results were obtained with 3-day-old seedlings. Conclusion, Recommendation and Outlook  (a) Sunflower and sorghum are good candidates to remediate As-polluted soils. (b) Phytoremediation can be improved with IAA or 2,4-D. (c) Mixed cropping of sorghum and sunflower may be another way of improving phytoremediation.     

Protein biofortified sorghum: effect of processing into traditional african foods on their protein quality

Protein biofortification into crops is a means to combat childhood protein-energy malnutrition (PEM) in developing countries, by increasing the bioavailability of protein in staple plant foods and ensuring sustainability of the crop. Protein biofortification of sorghum has been achieved by both chemically induced mutation and genetic engineering. For this biofortification to be effective, the improved protein quality in the grain must be retained when it is processed into staple African foods. Suppression of kafirin synthesis by genetic engineering appeared to be superior to improved protein digestibility by chemical mutagenesis, because both the lysine content and protein digestibility were substantially improved and maintained in a range of African foods. For the genetically engineered sorghums, the protein digestibility corrected amino acid score was almost twice that of their null controls and considerably higher than the high protein digestibility sorghum type. Such protein biofortified sorghum has considerable potential to alleviate PEM.     

Gluten-free sorghum bread improved by sourdough fermentation: biochemical, rheological, and microstructural background

This study was conducted to improve the quality and theoretical understanding of gluten-free sorghum bread. The addition of 2% hydroxypropyl methylcellulose improved bread based on 105% water, 70% sorghum flour, and 30% potato starch. Nevertheless, a flat top and tendency toward a hole in the crumb remained. Sourdough fermentation of the total sorghum flour eliminated these problems. Size-exclusion high-performance liquid chromatography demonstrated that during sourdough fermentation, proteins from the dough liquid were degraded to peptides smaller than kafirin monomers (<19 kDa). Laser scanning confocal microscopy showed aggregated protein in bread crumb without sourdough fermentation, whereas with sourdough fermentation, only small isolated patches of protein bodies embedded in matrix protein remained. In oscillatory temperature sweeps, sourdough fermentation caused a significantly higher resistance to deformation (|G*|) after gelatinization of the above batter relative to batters without sourdough. Results suggest that a strong starch gel, without interference of aggregated protein, is desirable for this type of bread.     

等离子体对鹰嘴豆分离蛋白溶解性和乳化特性的影响

鹰嘴豆作为植物蛋白的优质来源,营养价值高,但功能性质较差无法满足现代食品工业需求。该研究利用介质阻挡放电（Dielectric Barrier Discharge,DBD）等离子体对鹰嘴豆分离蛋白（Chickpea Protein Isolates,CPI）进行改性处理,研究不同处理时间（0、1、2、3、4 min）对CPI溶解性、乳化特性、结构的影响及其之间的相关性。结果表明：经等离子体处理后,鹰嘴豆分离蛋白溶液的pH值降低,电导率增加。溶解性、乳化活性和乳化稳定性得到显著的改善（P < 0.05）。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析表明等离子体处理并未改变CPI的组成成分及种类,但7S和11S等主要亚基条带强度增加。等离子体处理后α-螺旋含量、自由巯基含量和表面疏水性显著增加（P < 0.05）,无规卷曲含量降低（P < 0.05）,表明蛋白的高级结构发生改变。扫描电镜显示随着处理时间的延长,样品的尺寸减小,表面结构变得更为松散。利用Pearson相关性分析和主成分分析表明,不同处理时间后,蛋白结构的变化与功能性质的改善呈现较强的相关性。等离子体处理4 min后,CPI的溶解性及乳化特性达到较优效果,研究结果可为开发利用鹰嘴豆分离蛋白和指导实际生产实践提供技术支持。     

低温等离子快速提高糖基化花生分离蛋白溶解性及乳化性

为进一步提高花生蛋白的溶解特性,扩大花生蛋白在食品工业中的应用。采用低温等离子（Non-Thermal Plasma,NTP）诱导花生分离蛋白-葡聚糖（Peanut Protein Isolate-Dextran,PPI-Dex）湿法糖基化反应,研究NTP在处理0、0.5、1.5、2.0 、3.0 min的情况下,反应时间对花生分离蛋白与葡聚糖糖基化反应的影响。在低温等离子处理功率为70 W,反应液温度为60 ℃的状态下,随着NTP处理时间的延长,PPI-Dex的接枝度增加,在处理时间为1.5 min时,PPI-Dex接枝度达最大为21.62%,与超声波接枝PPI-Dex需要40 min,传统湿接枝需要24 h相比,缩短了接枝时间。PPI-Dex接枝后,接枝物溶解度和乳液稳定性显著增强,与未接枝相比,溶解度提高了22.28%。通过测定其分子量、氨基酸含量、红外图谱及表面疏水性变化分析NTP处理对花生分离蛋白结构影响。分析结果表明,NTP 处理1.5 min后,花生分离蛋白与葡聚糖发生糖基化反应形成偶联物,偶联物中羟基特征峰3 000～3 500 cm-1及1 000～1 260 cm-1的吸光度与未处理时相比增加,赖氨酸和苯丙氨酸相对含量显著降低（P<0.05）;同时,α-螺旋含量降低,β-折叠向β-转角转变,蛋白的有序结构被破坏,结构变松散,PPI构型向亲水型转变;接枝物的表面疏水性指数降低。花生分离蛋白与葡聚糖发生糖基化反应,反应位点可能为Lys和Phe。结果表明,低温等离子处理是一种快速促进蛋白与多糖接枝的有效方法。     

Evaluation of the simultaneous effects of processing parameters on the iron and zinc solubility of infant sorghum porridge by response surface methodology

The purpose of this study was to improve the micronutrient quality of indigenous African infant flour using traditional techniques available in the region. Response surface methodology was used to study the effect of duration of soaking, germination, and fermentation on phytate and phenolic compounds (PC), pH, viscosity, and the in vitro solubility (IVS) of iron and zinc in infant sorghum flour. The phytate and the PC concentrations of the flour were significantly modified as a result of the duration of germination and fermentation and their mutual interaction. These modifications were accompanied by a significant increase in % IVS Zn after 24 h of sprouting. Except for the interaction of soaking and fermentation, none of the processing parameters exerted a significant effect on the % IVS Fe. The viscosity of the porridge prepared with the flour decreased significantly with the duration of germination, making it possible to produce a porridge with high energy and nutrient density. The use of germination in combination with fermentation is recommended in the processing of cereals for infant feeding in developing countries.     

Determination of crude protein in animal feeds, using block digestion followed by steam distillation: collaborative study.

A method consisting of digesting animal feeds in a block digestor and determining ammonia by steam distillation followed by titration has been evaluated and compared with the official final action Kjeldahl method, 7.016. Fifteen laboratories analyzed 5 feed samples and lysine monohydrochloride. Statistical analysis showed that results from the 2 methods were comparable. The distillation technique has been adopted as official first action as an alternative technique for ammonia determination from the digest of the official final action block digestor method, 7.B11.