Bioconjugation of neutral protease on silk fibroin nanoparticles and application in the controllable hydrolysis of sericin

Bombyx mori silk fibroin is a protein-based macromolecular biopolymer with remarkable biocompatibility. Silk fiber was degummed and subjected to a series of treatments, including dissolution and dialysis, to yield an aqueous solution of silk fibroin, which was introduced rapidly into excess acetone to produce crystalline silk fibroin nanoparticles (SFNs). The SFNs were conjugated covalently with a neutral protease (NP) using glutaraldehyde as the cross-linking reagent. The objective of this study was to determine the optimal conditions for biosynthesis of the SFN-NP bioconjugates. First, SFN-NP was obtained by covalent cross-linking of SFN and NP at an SFN/NP ratio of 5-8 mg:1 IU with 0.75% glutaraldehyde for 6 h at 25 °C. When adding 50 IU of the enzyme, the residual activity of biological conjugates was increased to 31.45%. Studies on the enzyme activity of SFN-NP and its kinetics showed that the stability of SFN-NP bioconjugates was greater than that of the free enzyme, the optimum reactive temperature range was increased by 5-10 °C, and the optimum pH value range was increased to 6.5-8.0. Furthermore, the thermal stability was improved to some extent. A controlled hydrolysis test using the poorly water-soluble protein sericin as a substrate and SFN-NP as the enzyme showed that the longer the reaction time (within 1 h), the smaller the molecular mass (<30 kDa) is of the sericin peptide produced. The SFN-NP bioconjugate is easily recovered by centrifugation and can be used repeatedly. The highly efficient processing technology and the use of SFN as a novel vector for a protease has great potential for research and the development of food processing.     

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.     

Antioxidant and antiproliferative activities of loach ( Misgurnus anguillicaudatus ) peptides prepared by papain digestion

Loach protein hydrolysates (LPH) prepared by papain digestion were fractionated into four fractions, LPH-I (MW > 10 kDa), LPH-II (MW = 5-10 kDa), LPH-III (MW = 3-5 kDa), LPH-IV (MW < 3 kDa), and the in vitro antioxidant and antiproliferative (anticancer) activities of all fractions were determined. LPH-IV showed the lowest IC(50) value (16.9 ± 0.21 mg/mL) for hydroxyl radical scavenging activity and the highest oxygen radical scavenging capacity (ORAC) value (reaching 215 ± 5.9 mM Trolox/100 g loach peptide when the concentration was 60 μg/mL). Compared with other fractions, LPH-IV also exhibited stronger antiproliferative activity for human liver (HepG2), breast (MCF-7), and colon (Caco-2) cancer cell lines in a dose-dependent manner. When the protein concentration was 40 mg/mL, the HepG2 and MCF-7 cell proliferation of LPH-IV reached 7 and 4%, respectively, with no significant difference from those of LPH (8 and 7%, p > 0.05), with significantly less growth than those of LPH-I, LPH-II, and LPH-III, respectively (p < 0.05). The Caco-2 colon cell proliferation of LPH-IV was 12.8- and 8.7-fold smaller than those of LPH-I and LPH-II, respectively (p < 0.05). All of the fractions had a greater ability to inhibit Caco-2 colon cancer cell proliferation than to inhibit HepG2 liver cancer and MCF-7 breast cancer cell proliferation. The ORAC values of most of the fractions correlated (R(2) > 0.86, p < 0.01) with the antiproliferative activity of the three cancer cell lines, suggesting that higher antioxidant activity leads to better antiproliferative activity. However, further mechanistic and human clinical studies of the anticancer activity of loach protein hydrolysate fractions are needed.     

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.     

Solubilized wheat protein isolate: functional properties and potential food applications

Solubility, foaming capacity/stability, water holding and fat absorption capacities, and emulsifying capacity/stability of a solubilized wheat protein isolate (SWPI) were compared with those of commercial protein, that is, sodium caseinate (NaCAS), dried egg white (DEW), nonfat dry milk (NFDM), and soy protein isolate (SPI). SWPI was highly soluble at pH 6.5-8.5. Foaming capacity of SWPI was superior to those of SPI, NFDM, and DEW, and its foaming stability was similar to those of the commercial proteins. Foaming properties of SWPI were greatly improved in the presence of 0.5% (w/v) CaCl(2). Water holding capacity of SWPI was greater than that of NaCAS, NFDM, and DEW, whereas its fat absorption capacity was comparable to that of SPI, NaCAS, and DEW. SWPI exhibited emulsifying properties similar to those of SPI. SWPI was incorporated at 5, 10, 15, or 20% into ice cream, chocolate chip cookies, banana nut muffins, and hamburger patties. Products containing <5% SWPI were acceptable to consumers.     

Biochemical and functional properties of Atlantic salmon (Salmo salar) muscle proteins hydrolyzed with various alkaline proteases

Protein hydrolysates (5, 10, and 15% degrees of hydrolysis) were made from minced salmon muscle treated with one of four alkaline proteases (Alcalase 2.4L, Flavourzyme 1000L, Corolase PN-L, and Corolase 7089) or endogenous digestive proteases. Reaction conditions were controlled at pH 7.5, 40 degrees C, and 7.5% protein content, and enzymes were added on the basis of standardized activity units (Azocoll units). Proteases were heat inactivated, insoluble and unhydrolyzed material was centrifuged out, and soluble protein fractions were recovered and lyophilized. Substrate specificities for the proteases was clearly different. Protein content for the hydrolysates ranged from 71.7 to 88.4%, and lipid content was very low. Nitrogen recovery ranged from 40.6 to 79.9%. The nitrogen solubility index was comparable to that of egg albumin and ranged from 92.4 to 99.7%. Solubility was high over a wide range of pH. The water-holding capacity of fish protein hydrolysates added at 1.5% in a model food system of frozen minced salmon patties was tested. Drip loss was on average lower for the fish protein hydrolysates than for egg albumin and soy protein concentrate, especially for Alcalase hydrolysates. Emulsification capacity for fish protein hydrolysates ranged quite a bit (75-299 mL of oil emulsified per 200 mg of protein), and some were better than soy protein concentrate (180 mL of oil emulsified per 200 mg of protein), but egg albumin had the highest emulsifying capacity (417 mL of oil emulsified per 200 mg of protein). Emulsification stability for fish protein hydrolysates (50-70%) was similar to or lower than those of egg albumin (73%) or soy protein concentrate (68%). Fat absorption was greater for 5 and 10% degrees of hydrolysis fish protein hydrolysates (3.22-5.90 mL of oil/g of protein) than for 15% hydrolysates, and all had greater fat absorption than egg albumin (2. 36 mL of oil/g of protein) or soy protein concentrate (2.90 mL of oil/g of protein).     

Stabilization of oil-in-water emulsions by cod protein extracts

The ability of two protein fractions extracted from cod to form and stabilize oil-in-water emulsions was examined: a high salt extracted fraction (HSE protein) and a pH 3 acid extracted fraction (AE protein). Both fractions consisted of a complex mixture of different proteins, with the predominant one being myosin (200 kDa). The two protein fractions were used to prepare 5 wt % corn oil-in-water emulsions at ambient temperature (pH 3.0, 10 mM citrate-imidazole buffer). Emulsions with relatively small mean droplet diameters (d(3,2) < 1 microm) and good creaming stability (> 9 days) could be produced at protein concentrations > or =0.2 wt % for both fractions. The isoelectric point of droplets stabilized by both protein fractions was pH approximately 5. The emulsions were stable to droplet flocculation and creaming at relatively low pH (< or =4) and NaCl concentrations (< or =150 mM) when stored at room temperature. In the absence of salt, the emulsions were also stable to thermal treatment (30-90 degrees C for 30 min), but in the presence of 100 mM NaCl droplet flocculation and creaming were observed in some of the emulsions, particularly those stabilized by the AE fraction. The results suggest that protein fractions extracted from cod can be used as emulsifiers to form and stabilize food emulsions.     

Preparation and Functional Properties of Gluten Hydrolysates with Wheat‐Bug (Eurygaster spp.) Protease

Suni‐bug (Eurygaster spp.) enzyme was partially purified from bug‐damaged wheat and used to prepare gluten hydrolysates at 3% and 5% degree of hydrolysis (DH). Functional properties of gluten and gluten hydrolysates were determined at 0.2% (w/v) protein concentration and pH 2–10. Gluten solubility after enzymatic hydrolysis increased significantly (P < 0.05) up to 89.1, 89.6, and 95.0% at pH 7, 8, and 10, respectively. Emulsion stability (ES) of gluten hydrolysates improved at neutral and alkaline pH (P < 0.05) and emulsifying capacity (EC) increased significantly (P < 0.05) except at pH 10. Foaming capacity (FC) values of gluten hydrolysates were significantly higher (P < 0.05) at pH 6, 7, 8; foam stability (FS) values of gluten hydrolysates were significantly higher (P < 0.05) at pH 6 and 7. Enzymatic modification of gluten by wheat‐bug enzyme resulted in hydrolysates with higher antioxidant activity compared to gluten. Significant correlations (P < 0.001) were found between solubility and EC, ES, FC, and FS values of gluten and its hydrolysates with 3% and 5% DH.     

海藻肥对菜心抗旱性的影响及其机理探究

  【目的】  结合田间试验和盆栽试验,研究海藻肥对干旱胁迫条件下菜心产量和品质的影响,探讨其提高菜心抗旱性可能存在的机理,为海藻肥在叶菜上的应用提供理论支撑。  【方法】  1) 田间试验以‘碧清菜心’为材料,于2017年分别在广州 (华南主产区) 和宁夏 (供港澳有机蔬菜种植基地) 开展,土壤水分设为正常供水 (70%～75%田间土壤最大持水量) 和干旱 (50%～55%田间土壤最大持水量) 两个水平,设置清水对照 (CK)、海藻提取物 (SE)、水溶化肥 (NPK)、海藻肥 (NPK+SE) 4个处理。在菜心移栽7、14和21天时进行冲施,每次用量10 L/m2。移栽28天后收获,测定海藻肥对菜心经济产量和品质的影响。2) 盆栽试验于2018年在华南农业大学遮荫网室开展,正常供水只设清水对照 (CK),干旱条件下施肥处理同田间试验,施肥量改为100 mL/株。在移栽后14、21和28天时,测定菜心最大叶长、最大叶宽、株高,移栽后28天（收获时）测定叶片相对含水量,叶绿素和自由基含量,抗氧化酶活性和养分吸收量。3) 利用不同孔径的醋酸纤维超滤膜将海藻提取物分为分子量 > 10 kDa、5～10 kDa、3～5 kDa和 < 3 kDa等4种海藻提取物组分,测定其体外自由基清除能力及其对菜心抗旱性的影响。  【结果】  1) 田间试验结果表明,广州试验点正常供水条件下,与NPK处理相比,NPK+SE处理的菜心经济产量差异不显著,但干旱条件下显著增产;宁夏试验点在干旱和正常供水条件下,NPK+SE处理的菜心经济产量均显著高于NPK处理。广州试验点干旱和正常供水条件下,NPK+SE处理的各品质指标均高于NPK处理。宁夏试验点正常供水条件下,NPK+SE处理的菜心可溶性糖和可溶性蛋白含量较CK增加显著,而干旱条件下所有品质指标均显著增加。2) 盆栽试验结果显示,干旱条件下,SE处理的部分菜心品质指标值高于正常供水对照。收获时与NPK处理相比,NPK+SE处理显著提高了菜心株高、最大叶长和叶宽,鲜重显著增加12.60%,菜心叶绿素含量提高了10.24%,抗氧化酶活性提高27.84%～43.40%,叶片自由基含量降低了24.88%～41.56%。此外,NPK+SE处理的菜心氮、磷和钾吸收量分别较NPK处理增加了14.48%、16.41%和35.37%。3) 4个分子量不同的海藻提取物组分中,主要活性成分海藻酸、褐藻多酚、甘露醇在 < 3 kDa组分中的含量高于其他3个分子量组分。对超氧阴离子和羟基自由基的清除能力由大到小依次为< 3 kDa、> 10 kDa、5～10 kDa、3～5 kDa,以 < 3 kDa组分对干旱条件下菜心的促生作用最强。  【结论】  两个试验点的结果都表明,海藻提取物与水溶肥配合冲施可以显著提高菜心的经济产量和品质,干旱条件下的效果更显著。海藻提取物能够提高菜心的叶绿素含量、降低叶片自由基累积、增加氮磷钾的吸收,从而提高菜心抗旱性。< 3 kDa海藻提取物中的活性成分含量最高,因此,分子量 < 3 kDa的海藻提取物的自由基清除能力最强,对菜心的抗旱性提高效果最好。     

Enzymatic production of xylooligosaccharides from cotton stalks

Xylooligosaccharide (XO) production was performed from xylan, which was obtained by alkali extraction from cotton stalk, a major agricultural waste in Turkey. Enzymatic hydrolysis was selected to prevent byproduct formation such as xylose and furfural. Xylan was hydrolyzed using a commercial xylanase preparation, and the effects of pH, temperature, hydrolysis period, and substrate and enzyme concentrations on the XO yield and degree of polymerization (DP) were investigated. Cotton stalk contains about 21% xylan, the composition of which was determined as 84% xylose, 7% glucose, and 9% uronic acid after complete acid hydrolysis. XOs in the DP range of 2-7 (X6 approximately X5>X2>X3) were obtained with minor quantities of xylose in all of the hydrolysis conditions used. Although after 24 h of hydrolysis at 40 degrees C, the yield was about 53%, the XO production rate leveled off after 8-24 h of hydrolysis. XO yield was affected by all of the parameters investigated; however, none of them affected the DP of the end product significantly, except the hydrolysis period. Enzyme hydrolysis was maintained by the addition of fresh substrate after 72 h of hydrolysis, indicating the persistence of enzyme activity. The optimal hydrolysis conditions were determined as 40 degrees C, pH 5.4, and 2% xylan. The obtained product was fractionated via ultrafiltration by using 10, 3, and 1 kDa membranes. Complete removal of xylanase and unhydrolyzed xylan was achieved without losing any oligosaccharides having DP 5 or smaller by 10 kDa membrane. After a two-step membrane processing, a permeate containing mostly oligosaccharides was obtained.     

A novel angiotensin I converting enzyme inhibitory peptide from Alaska pollack (Theragra chalcogramma) frame protein hydrolysate

Alaska pollack frame protein, which is normally discarded as an industrial byproduct in the processing of fish in plants, was hydrolyzed with pepsin. This was fractionated into five major types of Alaska pollack frame protein hydrolysates (APH-I, 10-30 kDa; APH-II, 5-10 kDa; APH-III, 3-5 kDa; APH-IV, 1-3 kDa; and APH-V, below 1 kDa) using an ultrafiltration membrane bioreactor system. Angiotensin I converting enzyme (ACE) inhibitory activities of the fractionated hydrolysates were investigated, and the fraction that exhibited the highest ACE inhibitory activity was further purified using consecutive chromatographic methods on SP-Sephadex C-25 column, Sephadex G-25 column, and high-performance liquid chromatography (HPLC) on an octadecylsilane column. Finally, we purified a novel ACE inhibitory peptide with an IC50 value of 14.7 microM, and the sequence of the peptide was Phe-Gly-Ala-Ser-Thr-Arg-Gly-Ala. In addition, the ACE inhibition pattern of the peptide was found to be noncompetitive.     

Photochemical Degradation of Organic Pollutants in Solutions of Soil Humic Acids

Eurasian Soil Science - Chernozem humic acid (HA) fractions with nominal molecular sizes (MS) &gt;100, 30–100, 5–30 and &lt;5 kDa were obtained, using a combination of...     

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。     

Emulsifying and Foaming Properties of Okara Protein Hydrolysates

Okara is a low‐value coproduct of soy milk production. Its dry matter contains 25–30% protein that is of high nutritive quality, has an excellent efficiency ratio, and thus holds promise for applications in food systems. However, okara protein has low solubility. We here optimized its extraction and isolation from okara by using dilute sodium hydroxide and subsequent isoelectric precipitation. The obtained okara protein isolate (OPI) was hydrolyzed with different enzymes into a range of hydrolysates with different degrees of hydrolysis. Most hydrolysates had better emulsifying activity and produced more stable emulsions than OPI. In contrast, hydrolysis had no positive effect on foam‐forming and foam‐stabilizing activity of OPI proteins. Hydrolysis of OPI enhances the emulsifying capacity of the proteins. Furthermore, the emulsifying and foam‐forming capacities of most of the OPI hydrolysates were similar to or even better than those of the commercial (soy) protein hydrolysates used in this study.     

Antioxidant activity and emulsion-stabilizing effect of pectic enzyme treated pectin in soy protein isolate-stabilized oil/water emulsion

The antioxidant activity of pectic enzyme treated pectin (PET-pectin) prepared from citrus pectin by enzymatic hydrolysis and its potential use as a stabilizer and an antioxidant for soy protein isolate (SPI)-stabilized oil in water (O/W) emulsion were investigated. Trolox equivalent antioxidant capacity (TEAC) was found to be positively associated with molecular weight (M(w)) of PET-pectin and negatively associated with degree of esterification (DE) of PET-pectin. PET-pectin (1 kDa and 11.6% DE) prepared from citrus pectin after 24 h of hydrolysis by commercial pectic enzyme produced by Aspergillus niger expressed higher α,α-diphenyl-β-picrylhydrazyl (DPPH) radical scavenging activity, TEAC, and reducing power than untreated citrus pectin (353 kDa and 60% DE). The addition of PET-pectin could increase both emulsifying activity (EA) and emulsion stability (ES) of SPI-stabilized O/W emulsion. When the SPI-stabilized lipid droplet was coated with the mixture of PET-pectin and pectin, the EA and ES of the emulsion were improved more than they were when the lipid droplet was coated with either pectin or PET-pectin alone. The amount of secondary oxidation products (thiobarbituric acid reactive substances) produced in the emulsion prepared with the mixture of SPI and PET-pectin was less than the amount produced in the emulsion prepared with either SPI or SPI/pectin. These results suggest that PET-pectin has an emulsion-stabilizing effect and lipid oxidation inhibition ability on SPI-stabilized emulsion. Therefore, PET-pectin can be used as a stabilizer as well as an antioxidant in plant origin in SPI-stabilized O/W emulsion and thus prolong the shelf life of food emulsion.     

Assessment of the production of antioxidants from winemaking waste solids

Winemaking waste solids (WS, resulting from red grapes after fermentation and distillation to recover spirits) were subjected to various processing schemes for isolating fractions with antioxidant activity. The liquors entrapped in WS as received were separated by pressing and freeze-dried to yield a fraction with antioxidant activity (measured as DPPH radical scavenging capacity) comparable to those of synthetic antioxidants. A second approach based on the direct processing of raw WS in sulfuric acid medium under fixed operational conditions and further extraction of hydrolysis liquors with ethyl acetate enabled the isolation of a fraction with higher antioxidant ability at an improved yield. The most favorable approach started with a washing stage leading to liquors (which were directly freeze-dried to yield 1.20 g of extract/100 g of oven-dry WS and presented an EC50 of 0.41 g of extract/L) and washed solids, which were dried and subjected to hydrolytic processing (i) with water as a reactive in an autocatalyzed reaction (autohydrolysis) or (ii) with sulfuric acid solutions to give an ethyl acetate-soluble fraction with improved antioxidant properties (EC50 in the range of 0.18-0.40 g/L). Samples from washing liquors and processing of washed solids in aqueous medium were subjected to chromatographic fractionation and analysis to give isolates with remarkable antioxidant activity (with EC50 as low as 0.07 g/L) and to identify their major components.     

Chemical heterogeneity of humic substances: characterization of size fractions obtained by hollow-fibre ultrafiltration

To investigate the chemical heterogeneity of humic substances in relation to molecular size, fulvic and humic acids were extracted and purified from the surface horizon of a Humic Gleysol in northern Switzerland. A fractionation scheme using hollow‐fibre ultrafiltration cartridges was developed and used to obtain four size fractions of the humic acid with nominal molecular weight ranges > 300 kDa, 100–300 kDa, 30–100 kDa, and 10–30 kDa. The fulvic acid and all humic acid fractions were characterized by size exclusion chromatography, elemental analysis (C, H, N, S), as well as spectroscopic techniques including UV‐VIS, CP‐MAS ¹³C‐NMR, FT‐IR, and fluorescence spectroscopy. Clear chemical differences between the humic acid size fractions were observed. Smaller size fractions of the soil humic acid contained more chargeable functional groups and a larger percentage of aromatic carbon than the larger size fractions. Conversely, the percentage of aliphatic carbon increased with increasing apparent molecular weight. The chemical composition of the smallest humic acid fraction differed clearly from the fulvic acid fraction, despite similar apparent molecular size and carboxyl carbon content. Small humic acids contained much more aromatic carbon and less aliphatic carbon than the fulvic acid fraction. Apparently, humic size fractions differ in their chemical composition, which can have important implications for their environmental behaviour.     

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.     

Chemical characteristics and influences of two fractions of Chinese lignite humic acids on urease

Soil urease is crucial for the nitrogen cycle and its association with humic acids (HAs) is a fundamental requirement for its stability. In this work, the chemical characteristic of two HA fractions (HA1, ≥50 kDa; HA2, 10–50 kDa) extracted from lignite was evaluated, and their effects on the activity and stability of Jack Bean urease were also studied. HA1 and HA2 exhibited different structural properties in the micro-FT-IR and ¹³C NMR spectra and influences on urease stability during 12 days of incubation: HA1 stabilized the urease activity. After 12 days, the residual activity of urease, at pH 6.0, 7.0 and 8.0, was 2.1, 2.6 and 3.9 times higher in the treatment of HA1-urease than in the free urease, respectively. With pH values increasing, the stability of free urease decreased and that of HA1-urease increased, which indicated that HA1 improved the stability of urease in the solution, especially at the alkaline condition.     

Antioxidative properties of press juice from herring (Clupea harengus) against hemoglobin (Hb) mediated oxidation of washed cod mince

The antioxidative effect of herring (Clupea harengus) light muscle press juice (PJ) against hemoglobin-(Hb-) mediated oxidation of washed cod mince during ice storage was tested. The PJ was fractionated into low-molecular-weight (LMW; <1 [corrected] kDa) and high-molecular-weight (HMW; >1, >3.5, and > 50 kDa) fractions; it was preheated (10 min, 100 degrees C) and tested with or without removing heat coagulated proteins. Its antioxidative effect was compared with that given by endogenous levels of two tentative antioxidant candidates: ascorbic acid and uric acid. Oxidation was followed by determining rancid odor, peroxide value, and redness. Whole herring PJ and the LMW-PJ fraction significantly (p < 0.001) extended the oxidation lag phase of controls, from 2 up to 8 and 7 days, respectively. The HWM-PJ fractions were significantly (p < 0.05) less efficient than the whole and LMW-PJ samples, giving only 3.5-4.5 days of lag phase. Heat-treated PJ, with and without the heat-coagulated proteins, gave 7 and 5 days of oxidation lag phase, respectively. Heating different batches of the LMW-PJ fraction grouped the results into two categories: one where heating almost fully destroyed the antioxidative activity (fractions prepared from spring-caught herring) and another where heating had no or a minor effect (fractions prepared from fall-caught herring). The spring LMW-PJ had low ascorbic acid levels (18-42.6 microM), and 50-100% were destroyed by the heating. In fall LMW-PJ, the levels were 76.2-137.6 microM, and only 43-51% were destroyed. Ascorbic acid fortification of heated spring LMW-PJ to reach the levels found in the corresponding unheated spring LMW-PJ sample and the heated fall LMW-PJ gave back most of the antioxidative activity, which proved an important role of ascorbic acid for the antioxidative activity of LMW-herring PJ. This conclusion is drawn despite the fact that pure solutions with endogenous levels of ascorbic acid (giving 8.4-19.6 microM in final model) only very slightly delayed Hb-mediated oxidation of the washed cod mince.