Effects of Extraction Strategies on Yield, Physicochemical and Antioxidant Properties of Pumpkin Seed Oil.

This study investigated the effects of three extraction methods, including cold pressing (CP), microwave pretreatment pressing (MP), and supercritical fluid extraction (SFE), on the yield, physicochemical properties, bioactive compounds content, and antioxidant properties of pumpkin seed oil (PSO). Furthermore, the correlation between bioactive compounds and the antioxidant properties of PSO was determined. The results revealed that the yield of PSO extracted using the three methods was in the order of SFE > MP > CP. Additionally, the PSO generated by SFE showed the highest unsaturated fatty acid content, followed by MP and CP. Additionally, MP-PSO exhibited the highest acid value and saponification value, while SFE-PSO displayed the highest moisture content, peroxide value, and iodine value. Moreover, the PSO generated by MP demonstrated superior antioxidant properties compared to that of PSOs from CP and SFE in the oxidation induction, DPPH, FRAP, and ABTS tests. Finally, the correlation analysis revealed that specific types of bioactive compounds, such as ß-sitosterol and γ-tocopherol, were highly correlated with the antioxidant properties of PSOs. Consequently, this study provides comprehensive knowledge regarding PSO extraction, physicochemical properties, bioactive compound extraction, and the correlated antioxidant properties.

Selection of Salt-Tolerance and Ester-Producing Mutant Saccharomyces cerevisiae to Improve Flavour Formation of Soy Sauce during Co-Fermentation with Torulopsis globosa.

Soy sauce, as a traditional seasoning, is widely favoured by Chinese and other Asian people for its unique colour, smell, and taste. In this study, a salt-tolerance Saccharomyces cerevisiae strain HF-130 was obtained via three rounds of ARTP (Atmospheric and Room Temperature Plasma) mutagenesis and high-salt based screening. The ethanol production of mutant HF-130 was increased by 98.8% in very high gravity fermentation. Furthermore, ATF1 gene was overexpressed in strain HF-130, generating ester-producing strain HF-130-ATF1. The ethyl acetate concentration of strain HF-130-ATF1 was increased by 130% compared to the strain HF-130. Finally, the soy sauce fermentation performance of Torulopsis globosa and HF-130-ATF1 was compared with T. globosa, HF-130, HF-130-ATF1, and Torulopsis and HF-130. Results showed ethyl acetate and isoamyl acetate concentrations in co-fermentation of T. globosa and HF-130-ATF1 were increased by 2.8-fold and 3.3-fold, respectively. In addition, the concentrations of ethyl propionate, ethyl caprylate, phenylethyl acetate, ethyl caprate, isobutyl acetate, isoamyl alcohol, phenylethyl alcohol, and phenylacetaldehyde were also improved. Notably, other three important flavour components, trimethylsilyl decyl ester, 2-methylbutanol, and octanoic acid were also detected in the co-fermentation of T. globosa and HF-130-ATF1, but not detected in the control strain T. globosa. This work is of great significance for improving the traditional soy sauce fermentation mode, and thus improving the flavour formation of soy sauce.

Effects of Structural and Compositional Changes of Nanochloropsis oceania after Enzyme Treatment on EPA-Rich Lipids Extraction.

Improved methods for the extraction of eicosapentaenoic acid (EPA), an essential and economically important polyunsaturated fatty acid, are urgently required. However, lipid extraction rates using food-grade solvents such as ethanol are usually low. To improve the ethanol-based extraction rate, and to elucidate the relevant mechanisms, we used cellulase and laccase to treat powdered Nannochloropsis, one of the most promising microalgal sources of EPA. Cellulase and laccase synergistically increased lipid yields by 69.31% and lipid EPA content by 42.63%, by degrading the amorphous hemicellulose and cellulose, improving crystallinity, and promoting the release and extraction of lysodiacylglyceryltrimethylhomoserine. Scanning electron microscopy showed that cell morphology was substantially altered, with cell-wall rupture, loss of cell boundaries, and the release of intracellular substances. In conclusion, Nannochloropsis lipid yields may be directly linked to cell-wall hemicellulose structure, and enzymatic treatment to alter this may improve lipid yields.

Spectral noise-to-signal ratio priority method with application for visible and near-infrared analysis of whole blood viscosity.

Whole blood viscosity (WBV) is a group of important clinical indicators of cardio-cerebral vascular diseases. Existing detection methods for WBV are complex, making them inconvenient for large population screening. Blood viscosity is closely related to the deformability and aggregation of erythrocytes, which are associated with haemoglobin. Haemoglobin has obvious near-infrared (NIR) spectral absorption. Scattering occurs when NIR light enters a viscous blood sample, and its scattering degree is correlated with blood viscosity. Based on repeated spectral measurements and spectral similarity, spectral noise-to-signal ratio (NSR) was proposed to quantify the spectral scattering effect in the blood sample. A novel selection method of piecewise-continuous wavelengths, named NSR priority-partial least squares (NSRP-PLS), was proposed and applied for visible-NIR quantitative analysis of WBV with high, medium and low shear rates [WBV(H), WBV(M), WBV(L)]. Modelling was separately performed by gender to allow for systematic gender differences in blood viscosity. For the NIR-predicted and clinically measured values of the three WBV indicators in independent validation, the root mean square errors for prediction (SEP) were 0.498, 0.222 and 0.193 (mPa·s), respectively. And the correlation coefficients (RP) were 0.927, 0.934 and 0.927, respectively. Compared with the three current well-performing methods (MW-PLS, CARS-PLS and SPA-PLS), the proposed NSRP-PLS method achieved better predictive accuracy. Results indicated that visible-NIR spectroscopy combined with the NSRP-PLS method can be used for the quantitative analysis of WBV. The proposed analytical method is rapid, reagent-free and is scientific and meaningful for cardio-cerebral vascular diseases screening in large populations.

Characterization of a salt-tolerant aminopeptidase from marine Bacillus licheniformis SWJS33 that improves hydrolysis and debittering efficiency for soy protein isolate.

An aminopeptidase was isolated from the marine Bacillus licheniformis SWJS33 (BLAP) and purified. According to the tandem mass spectrometry, the enzyme displayed 11% amino acid identity with the aminopeptidase from Bacillus (gi|496687392). BLAP exhibited maximum activity at 60°C and pH 8.0-8.5 and had a molecular mass of 100kDa. The presence of NaCl enabled 50% improvement of enzyme activity with 10-15% NaCl being the best. The observed inactivation by EDTA and bestatin and activation by Co(2+) and Ag(+) indicated that the obtained enzyme was a metalloaminopeptidase. Such an aminopeptidase could further improve the hydrolysis degree of soy protein isolate hydrolysates catalyzed by papain, Alcalase 2.4L or Flavourzyme 500MG from 8.5%, 9.5% or 14.4-18.8%, 18.7% or 20.1%, respectively, while decreasing the bitter intensity score of the SPI hydrolysates catalyzed by Alcalase 2.4L from 3.6 to 0.4.

Evaluation of the Hydrolysis Specificity of an Aminopeptidase from Bacillus licheniformis SWJS33 Using Synthetic Peptides and Soybean Protein Isolate.

The substrate specificity of aminopeptidases has often been determined against aminoacyl-p-nitroanilide; thus, its specificity toward synthetic peptides and complex substrates remained unclear. The hydrolysis specificity of an aminopeptidase from Bacillus licheniformis SWJS33 (BLAM) was evaluated using a series of synthetic peptides and soybean protein isolate. The aminopeptidase showed high specificity for dipeptides with Leu, Val, Ala, Gly, and Phe at the N-terminus, and the specificity was significantly affected by the nature of the penultimate residue. In the hydrolysis of soy protein isolate, BLAM preferred peptides with Leu, Glu, Gly, and Ala at the N-terminus by free amino acid analysis and preferred peptides with Leu, Ala, Ser, Trp, and Tyr at the N-terminus by UPLC-MS/MS. The introduction of complex substrates provides a deeper understanding of the aminopeptidase's specificity, which can instruct the application of the enzyme in protein hydrolysis.

Purification and characterization of a new neutral metalloprotease from marine Exiguobacterium sp. SWJS2.

Among the protease-producing bacterial strains isolated from deep-sea sediments, SWJS2 was finally selected and identified as genus Exiguobacterium. Plackett-Burman and orthogonal array designs were applied to optimize the fermentation conditions, and the results are as follows: Glucose 5g, yeast extract 15g, glycerin 2g and CaCl2 ⋅2H2 O 0.5 g dissolved in 1 L artificial seawater; temperature 25 °C, original pH 7, inoculum rate 2%, seed age 12 H, loading volume 25 mL (250-mL Erlenmeyer flask), shaking speed 150 rpm, and fermentation time 44 H. The protease activity production was improved from about 80 to 660 U/mL under the optimized parameters. The protease was purified fourfold with specificity activity of 30,654.1 U/mg protein and a total yield of 16.2%. The protease exhibited the maximum activity at 40-45 °C and pH 7. Moreover, the enzyme activity was found to be inhibited by Cu(2+) , Ba(2+) , Cd(2+) , Hg(2+) , and Al(3+) at 5 mM, whereas it can be increased by Mg(2+) , Mn(2+) , and Ca(2+) at 0.5-5 mM. The enzyme was totally inactivated by 1 or 5 mM ethylenediaminetetraacetic acid but not by phenylmethanesulfonyl fluoride, tyrpsin inhibitor from Glycine max (STI), benzamidine, 5,5'-dithio-bis-(2-nitro benzoic acid), or pepstatin A, suggesting that it belonged to metalloprotease.

Evaluation of the hydrolysis specificity of protease from marine Exiguobacterium sp. SWJS2 via free amino acid analysis.

This study evaluates the hydrolysis specificity of newly developed protease from Exiguobacterium sp. SWJS2 (EP) based on the released free amino acid (FAA) during enzymolysis and through comparing with commercially available papain and Alcalase 2.4L. Results showed that EP had great potential in producing hydrolysates with better nutrition and less bitterness. The percentages of essential amino acids in the EP-treated Coilia mystus and soybean protein were 72 and 70 %, respectively. And, corresponding hydrophobic amino acids were 74 and 72 %, respectively (i.e. <60 % for papain- or Alcalase 2.4L-treated samples). The differences in FAA releasing rates between EP and the two commercial proteases suggested that EP could become a new commercial protease that offers different reaction rates and extends protease application scope. Track changes in FAA profiles throughout EP hydrolysis revealed that the release of each amino acid exhibited its distinct regularity as the hydrolysis proceeded, which also varied with the substrate proteins. EP hydrolysis of C. mystus and soybean protein led to greater production rates of Phe, Leu, Val, Ile and Ala, suggesting that EP might have higher preference to these amino acids.

GC/MS analysis of volatiles obtained by headspace solid-phase microextraction and simultaneous-distillation extraction from Rabdosia serra (MAXIM.) HARA leaf and stem.

Volatiles in Rabdosia serra were investigated by headspace solid phase microextraction (HS-SPME) and simultaneous-distillation extraction (SDE). The HS-SPME technique was previously evaluated to optimise sampling conditions. A total of 56 and 48 compounds including alcohols, aldehydes, hydrocarbons, ketones, carboxylic acid, ester, and aromatics were identified in leaf and stem by optimised HS-SPME method (CAR/PDMS fibre; incubation time, 10 min; extraction temperature, 50°C; extraction time, 40 min), respectively. 1-Octen-3-ol and (2E)-hexenal had significant contribution to R. serra aroma. Cluster analysis indicated that leaf and stem exhibited different volatile diversity. Air drying was favourable for the retention of the volatiles, while freeze- and sun-drying led to the loss of volatiles. SDE method preferred to the analysis of compounds with low volatility including fatty acids and esters. HS-SPME was a useful technique for the analysis of readily volatile components for the characteristics of R. serra aroma.

Structural characteristics of water-soluble polysaccharides from Rabdosia serra (MAXIM.) HARA leaf and stem and their antioxidant capacities.

Water-soluble polysaccharides of Rabdosia serra leaf and stem were fractionated by ultrafiltration and DEAE-Sepharose fast flow chromatogram to obtain water (RSLP-I and RSSP-I), 0.1M NaCl (RSLP-II and RSSP-II) and 0.2M NaCl (RSLP-III and RSSP-III) eluates. Their molecular weights were determined by high performance gel permeation chromatography. Monosaccharide composition analysis indicated that the water eluates comprised of rhamnose, arabinose, xylose, mannose, glucose and galactose. The ascending percentage of galactose and descending percentage of glucose in the eluates were observed with the increase of NaCl concentration. The branched RSLP-I and RSSP-I were composed mainly of →6)-Glcp-(1→, →6)-Galp-(1→, and →5)-Araf-(1→ residues with the ratio of 15.0:4.2:3.8 and 5.5:6.4:3.5, respectively. However, a low level of 1,6-linked glucosyl was observed in RSLP-II, RSSP-II, RSLP-III and RSSP-III. The structural characteristics were further analysed by infrared spectrophotometry. The purified leaf and stem polysaccharides possessed moderate antioxidant capacities.

Multiple headspace solid-phase microextraction using a new fiber for avoiding matrix interferences in the quantitative determination of ethyl carbamate in pickles.

Multiple headspace solid-phase microextraction (HS-SPME) using a novel fiber coated with anilino-methyl triethoxy silicane-methacrylic acid/terminated silicone oil has been introduced as a useful pretreatment technique coupled to gas chromatography-flame ionization detector for the detection of ethyl carbamate in pickles. Anilino-methyl triethoxy silicane and methacrylic acid are put into use simultaneously with the aim to increase the hydrogen interaction strength between ethyl carbamate and the coating. In addition, the new fiber exhibits high thermal stability, good reproducibility, and long lifetime. Extraction temperature, extraction time, amount of desiccant, and amount of sample were well optimized to guarantee the suitability of multiple HS-SPME. Significant matrix interference was observed among various types of pickles and the multiple HS-SPME procedure was proved to be effective in avoiding the matrix effect by a complete recovery of the analyte. The method showed satisfactory linearity (0.1-100 mg kg(-1)), precision (4.25%, n = 5), and detection limit (0.038 mg kg(-1)). The accuracy of the method was evaluated by comparison with standard addition method and the results were statistically equivalent. The study indicates that the multiple HS-SPME procedure is simple, convenient, accurate, and low-cost, and most of all, can be used for quantitative analysis in complex matrix without matrix effect.

Thermal inactivation kinetics of Rabdosia serra (Maxim.) Hara leaf peroxidase and polyphenol oxidase and comparative evaluation of drying methods on leaf phenolic profile and bioactivities.

Inactivation kinetics of peroxidase and polyphenol oxidase in fresh Rabdosia serra leaf were determined by hot water and steam blanching. Activation energy (52.30 kJ mol(-1)) of polyphenol oxidase inactivation was higher than that (20.15 kJ mol(-1)) of peroxidase. Water blanching at 90 °C or steam blanching at 100 °C for 90 s was recommended as the preliminary treatment for the retention of phenolics. Moreover, comparative evaluation of drying methods on the phenolics profiles and bioactivities of R. serra leaf were conducted. The results indicated that only intact leaf after freeze drying retained the initial quality. The sun- and air-dried leaves possessed identical phenolic profiles. The homogenised leaf (after freeze-drying) possessed a lower level of phenolics due to enzymatic degradation. Good antioxidant activities were detected for the sun- and air-dried leaves. There was insignificant difference in anti-tyrosinase and anti-α-glucosidase activities among sun-, air-, and freeze-dried leaves.