Adenosine cyclic phosphate with ultrasonic-assisted pectinase extraction alleviated allergic reactions in RBL-2H3 through inhibiting the influx of intracellular Ca^(2+)

Jujube contains abundant cyclic adenosine monophosphate(cAMP)and the ultrasonic-assisted pectinase extraction(UAPE)conditions for obtaining the maximum cAMP yield from jujube were optimized.Orthogonal array design was applied to evaluate the effects of 4 variables by UAPE on cAMP yield.The results showed that the optimal cAMP yield(783.0μg/g)was derived at ratio of liquid to solid 5 mL/g,ratio of pectinase to raw material 1.5%,time 60 min and temperature 40℃.Moreover,the effect of cAMP on the anti-allergic function of action induced by immunoglobulin E(IgE)and its meschanism was investigated through establishing the sensitized cell model in rat basophilic leukemia(RBL-2 H3)cells using dinitrophenylated(DNP)-bovine serum albumin(BSA)-IgE.The results showed that cAMP interfered with sensitized cells,effectively inhibited the occurrence of basophil degranulation in dose dependence,and significantly reduced the activity ofβ-hexosamindase(β-hex),at the optimal concentration of 50μg/mL.The level of anti-inflammatory factor interleukin-10(IL-10)was promoted and the content of pro-inflammatory factor tumor necrosis factor-α(TNF-α)was suppressed by cAMP.In addition,influx of intracellular Ca^(2+) was repressed effectively.Our results demonstrate that jujube cAMP regulated the cytokine balance in the allergy pathway through blocking the influx of extracellular Ca^(2+),with the prevention of allergy symptoms.

Polyphenols extracted from Aronia melanocarpa by combined enzymatic and ultrasonication and their antioxidant potential and antimicrobial activity

The extraction of polyphenols from Aronia melanocarpa was carried out using a combination of enzymatic and ultrasound.After single-factor and orthogonal design and experiment,the optimized polyphenol extraction conditions were 1%enzyme,1:40 material-to-liquid ratio,55℃,60 min ultrasonication,70%ethanol,and the final extraction amount was 88.634 mg/g,which displayed a 25.15%and 34.08%improvement compared with the single ultrasonication and enzymatic extraction methods,respectively.Significant antibacterial effects of polyphenols were shown against Staphylococcus aureus,Escherichia coli and Bacillus subtilis.Further antioxidation effects were evaluated,and the superoxide anion radical scavenging rate,hydroxyl radical scavenging rate and DPPH free radical scavenging rate reached 45.2%,83.5%and 85.4%,respectively.This combined enzymatic and ultrasonic extraction method exhibited the advantages of high extraction rate,saving solvent consumption and extraction time,but also provided a new method for the development and utilization of natural antimicrobial and antioxidant health products.

Antioxidant and hepatoprotective effects of Ajuga nipponensis extract by ultrasonic-assisted extraction

Objective: To investigate suitable condition for extraction of the active components from Ajuga nipponensis(A. nipponensis). Methods: Orthogonal experimental design was used to determine the optimal extraction parameters for ecdysterones and flavonoids. Finally, the hepatoprotective abilities of A. nipponensis extracts were evaluated by CCl_4-induced animal models. Results:Maximum yields of flavonoids(7.87±0.10) mg/g and ecdysterones(0.73±0.02) mg/g could be obtained when the extraction time was 50 min, the extraction temperature was 60 ℃, and the ratio of sample to 70%(v/v) ethanol was 1:20(w/w). The antioxidant property of A. nipponensis was correlated to the concentration of its extracts. At 5 mg/m L, A. nipponensisextract scavenged 84.8% of DPPH radical and had absorbance values of 2.43±0.04 reducing power. Upon CCl_4-induced liver injury, glutamic oxaloacetic transaminase and glutamic pyruvic transaminase decreased significantly after the mice were treated with A. nipponensis. Histological researches also explained that A. nipponensis reduced the extent of liver lesions induced by CCl_4. Conclusions: A. nipponensis exhibited potent antioxidant activity in chemical experimental models and hepatoprotective effect against CCl_4-induced liver damage.

Extraction process of chlorogenic acid in flos lonicerae by enzymatic treatment

A new method of extracting chlorogenic acid from flos lonicerae, and treating the materials with enzyme before being extracted by ethanol is developed, and the optimum conditions are also investigated in detail. Three important factors, enzyme dosage, treatment time and treatment temperature are adapted to optimize the extraction process. The experimental results show that the extract yield of flos lonicerae and chlorogenic acid can be obviously increased by the cellulase treatment, 61.5 mg chlorogenic acid is obtained from 1.00 g flos lonicerae at most. The optimal temperature of enzymatic treatment is 40 50 ℃. Compared with the use of single cellulase, the combined treatment of cellulase and pectinase increase the extract yield obviously but fail to improve that of chlorogenic acid.

Optimization of Ultrasonic-Assisted Extraction Process of Polysaccharides from American Ginseng and Evaluation of Its Immunostimulating Activity

Ultrasonic-assisted extraction （UAE） of American ginseng polysaccharides （AGP） was investigated using response surface methodology. Three-factor-three-level Box-Behnken design was employed to optimize the ultrasonic power, extraction time and ratio of water to raw material to obtain a high AGP yield. The analysis of variance and response surface plots indicated that ultrasonic power was the most important factor affecting the extraction yield. The optimal conditions were ultrasonic power 400 W, extraction time 71 min, and ratio of water to raw material 33 mL g-1. Under these conditions, the yield of AGP was 8.09%, which was agreed closely to the predicted value. Gas chromatography （GC） analysis showed that AGP was composed of arabinose, rhamnose, galactose, glucose, and galacturonic acid. Fourier transform infrared spectra revealed the general characteristic absorption peaks of AGP. In addition, AGP exhibited good immunostimulating activities by up-regulating the production of nitric oxide and cytokines. Compared with hot water extraction, UAE required shorter extraction time and gave a higher extraction yield, without changing the structure and immunostimulating activity of AGP. The results indicated that UAE could be an effective and advisable technique for the large scale production of plant polysaccharides.

Enzymatic Aqueous Extraction of Soybean Oil

The procedure of enzymatic aqueous extraction of soybean oil was assessed when two-step controlled enzymatic hydrolysis was applied. With aqueous extraction of soybean oil-containing protein, the highest yield of oil was 96.1% at the optimized conditions studied. Soybean oil-containing protein was hydrolyzed and resulted in releasing part of oil. The separated protein that contained 40% oil was enriched due to its adsorption capacity of released oil, the average oil extraction yeild reached 93.5%. Then the high oil content protein was hydrolyzed again to release oil by enzyme, the oil extraction yeild was 80.4%. As a result, high quality of soybean oil was obtained and the content of total oil yield was 74.4%.

Extraction Hydrolysates from Larimichthys Polyactis Swim Bladder Using Enzymatic Hydrolysis

As a kind of biopolymer,hydrolysates of fish swim bladder,safer than those of land mammals,are widely used in food,cosmetics as well as pharmaceutical and biomedical fields for their biocompatibility,biodegradability,and weak antigenicity.To enhance hydrolysate production,in this paper,the papain and alcalase hydrolysis processes of larimichthys polyactis swim bladder were optimized with orthogonal experiments.With 89.5%hydrolysate yield,the optimal processing conditions for alcalase were solid-liquid ratio of 1:30,enzyme concentration of 0.7%,and extraction time of 6 h.As for papain,under the optimal processing conditions:solid-liquid ratio of 1:20,enzyme concentration of 0.5%,and extraction time of 8 h,the hydrolysate yield was 65.1%.To obtain higher hydrolysate yields,the ultrasonic pretreatments were implemented before the optimal enzyme hydrolysis processes.With ultrasonic waves of 100 W for 50 min,the hydrolysate yields were increased 2.1%(alcalase)and 4.5%(papain),respectively.The Fourier Transform Infrared(FTIR)spectroscopic analysis revealed that the hydrolysates extracted by papain exist in triple-helical forms.The Ultra-Violet(UV)absorption spectra indicated that the aromatic amino acids in the hydrolysates had strong absorptions in the wavelength range of 240 nm–300 nm.The results of this research demonstrate that the alcalase hydrolysates have better solubility in water and the solution is more stable under ambient temperature.However,the hydrolysates extracted by papain have a gel property and are insoluble in weak acid at room temperature,which is more suitable for applications in feedstock of biomedical.

Ultrasound-Assisted Aqueous Enzymatic Extraction of Oil from Perilla(Perilla frutescens L.)Seeds

In the present study the effects of ultrasonic pretreatment and the types of enzyme on oil yield were investigated.The optimum ultrasonic pretreatment parameters were found to be 250 W of ultrasonic power,30 min of ultrasonic time,and 50℃of ultrasonic temperature.Five types of enzyme,Cellulase,Viscozyme L,Alcalase 2.4L,Protex 6L,and Protex 7L,were evaluated for their effectiveness in releasing oil from ultrasonic pretreated perilla seeds.The highest oil yield of 81.74%was observed in cellulase treated perilla seed samples.The physicochemical properties of the control,hexane,and enzyme extracted perilla seed oils were compared.No significant(P>0.05)differences were observed in iodine value,refractive index,unsaponifiable matter,saponification value,peroxide value,and acid value.

Effect of enzymatic extraction conditions on the yield and the polymerization degree of procyanidins

Grape seeds are rich sources of procyanidin(PCs)known for potential health benefi ts.In this study,PCs were extracted from defatted grape seeds by enzymatic method in which pectinase and cellulase were used.The enzyme extraction process was further optimized by single factor experiment and response surface methodology.The optimal conditions were as follows:ethanol concentration of 70%,extraction time of 70 min,extraction temperature of 35℃,liquid/solid ratio of 103:1(mL/g),pectinase/cellulase ratio of 1:1,enzyme/solid ratio of 1:314 w/w.Under the above conditions,the extraction yields and mean degree of polymerisation(mDP)of PCs reached 47.18 mg/g dry material weight and 11.2,respectively.Compared with other extraction methods,enzyme extraction can obtain PCs with higher yield and lower mDP.According to the antioxidant activity test,PCs extracts with lower mDP showed better ability to clear 1,1-Diphenyl-2-picrylhydrazyl radical(DPPH).Enzymatic extraction was an effi cient method to obtain oligomeric procyanidin which has stronger antioxidant activity.

Simplex - Centroid Mixture Design Applied to theAqueous Enzymatic Extraction of Fatty Acid - BalancedOil from Mixed Seeds

A one-step method was developed to extract oil from a mixture of soybeans,peanuts,linseeds,and tea seeds using an aqueous enzymatic method.The proportion of the four seeds was targeted in accordance with a fatty acid ratio of 0.27(SFA,saturated fatty acid(s))∶1(MUFA,monounsaturated fatty acid(s))∶1(PUFA,polyunsaturated fatty acid(s)),and the oil extraction yield was maximized by applying the simplex-centroid mixture design method.Three models were developed for describing the relationship between the proportion of the individual seeds in the mixture,the fatty acid ratio in the extracted oil,and the oil extraction yield,respectively.The developed models were then analyzed using an ANOVA and were found to fit the data quite well,with R2 values of 0.98,0.93,and 0.93,respectively.The three models were validated experimentally.The results indicated that the ratio of fatty acids in the oil ranged between 0.98 and 1.12(MUFA∶PUFA)and between 0.26 and 0.28(SFA∶MUFA),which were quite close to the target values of 1 and 0.27,respectively.The oil extraction yield of 62.13%was slightly higher than the predicted value(60.32%).

Research on Technology Development of Traditional Chinese Medicine Extraction Based on Patent Information

Objective To understand the development status of traditional Chinese medicine extraction technology,so as to provide a reference for enterprises and scientific research institutions to determine the direction of technological innovation.Besides,the government can formulate scientific and technological innovation strategies.Methods Based on patent information,statistical methods were applied to analyze the development of traditional Chinese medicine extraction technology.Then,the core technology was identified by constructing a patent citation network.Finally,the main path analysis method was applied to discover the technological evolution path.Results and Conclusion The extraction technology of traditional Chinese medicine is still in the growth stage.At present,its R&D is based on enterprises,supplemented by scientific research institutions.The core technologies are ultrasonic extraction and enzymatic extraction.The main path represented by enzymatic extraction technology reveals the evolution process of traditional Chinese medicine extraction technology.Based on this,relevant suggestions are put forward.Firstly,technological innovation cooperation should be strengthened to explore new models of achievement transformation and cooperation.Secondly,key enterprises of technology clusters should be cultivated to accelerate breakthroughs in core technologies.Thirdly,combined technologies can achieve high-efficiency enzyme-assisted extraction and open up new ways of extracting traditional Chinese medicine.Fourthly,traditional techniques should be improved to develop innovative modern techniques.

Assessment of Beauveria bassiana and Their Enzymatic Extracts against Metamasius spinolae and Cyclocephala lunulata in Laboratory

The application of enzymatic extracts and conidia of Beauveria bassiana in Metamasius spinolae and Cyclocephala lunulata was evaluated. The variables were mortality and time of death. In M. spinolae, mortality with extracts 29%, conidia 27% and the combination of both 31%, all had a time of death of four days. Although with different symptoms, used enzymatic extracts: contraction and softening of the joints;by conidia: mycelium in the joints;in the combination of conidia and enzymatic extracts: abundant aerial mycelium. In C. lunulata, 100% mortality in all treatments;Time of death: enzymatic extracts and extracts with conidia 1.2 days;conidia 2.8 days. Symptoms were different, enzymatic extracts: melanization and internal tissue lysis;enzymatic extract and conidia: mycelium emerged and melanization;conidia: mycelium emerged. Enzymatic extracts showed insecticidal activity in M. spinolae and C. lunulata. These results suggest the potential of enzymatic extracts as biocontrol agents to improve the use of entomopathogenic fungi.

Alleviation of Drought Stress in Wheat Using Exogenous Ulva prolifera Extract Produced by Enzymatic Hydrolysis

Drought is one of the major abiotic stresses that affect plant growth and reduce agricultural productivity.Use of algal extract as a biostimulant is gaining increased attention from researchers.This study aimed to investigate the potential of Ulva prolifera extract(UE)as a biostimulant when enzymatically extracted under conditions of water deficit.UE treatments(0.02%,0.06%,and 0.1%)significantly improved the shoot length,root length,and dry weight of roots after 120 h of drought stress relative to that in treatment with the negative control.An increase in catalase(CAT)and peroxidase(POD)activity was also observed that resulted in improved antioxidant capacity.Application of 0.1%UE reduced the malondialdehyde(MDA)content by 30.06%compared with that in the negative control.In addition,the soluble sugar and protein content in wheat treated with 0.1%UE was increased by 23.10%and 93.51%,respectively,resulting in adjustment of the osmotic pressure.Results suggest that UE could significantly enhance the drought tolerance of wheat.This study provides a basis for increasing the value of UE as a biostimulant.

Isolation and purification of carbazole contained in anthracene slag by extraction combined with medium pressure liquid chromatography

Ultrasonic-assisted extraction(UAE)combined with medium pressure liquid chromatography(MPLC)was designed for carbazole separation from anthracene slag(AS).The effects of liquid/solid ratio,temperature,and extraction times on carbazole separation were investigated.When using CC14 and ethyl acetate as extraction solvents and combining with MPLC,carbazole recovery and purity are 75.1%and 95.4%,respectively.The mechanism for carbazole separation were presumed by examining intermolecular interactions such as N-H…π,π-π,and C-Cl…πinteractions.These results demonstrate that UAE/MPLC has a considerable potential as a green and promising strategy for separating and purifying carbazole and other chemicals from AS.

Optimization of Extraction Process of Quercetin from Flos Sophorae Immaturus Using Orthogonal Design

[Objectives]This study aimed to optimize the ultrasonic-assisted extraction process of quercetin from Flos Sophorae Immaturus.[Methods]The natural product quercetin in Flos Sophorae Immaturus was extracted by ultrasonic-assisted method,and the content of quercetin in the extract was determined by ultraviolet spectrophotometry.The effect of ethanol concentration,solid/liquid ratio,temperature,time and ultrasonic power on the yield of quercetin in Flos Sophorae Immaturus was investigated by using single-factor experiments.Based on the results of single-factor experiments,an orthogonal experiment was designed to optimize the extraction process.The experimental data obtained were subjected to range analysis,analysis of variance and SSR test using SPSS 20.0 software to obtain the optimal extraction process.The results concluded were verified.[Results]The optimal ultrasonic-assisted extraction process for quercetin in Flos Sophorae Immaturus was as follows:temperature of 85℃,time of 30 min,solid/liquid ratio of 1∶20 g/mL,ethanol concentration of 50%.Under the optimal extraction conditions,the yield of quercetin in Flos Sophorae Immaturus was the highest.[Conclusions]This study provides a theoretical basis for the application of quercetin in Flos Sophorae Immaturus.

Enzyme from Banana(Musa sp.)Extraction Procedures for Sensitive Adrenaline Biosensor Construction

For building a biosensor, at least two enzyme sources can be employed. The pure enzyme has features with better selectivity and low stability. Crude extract presents better stability and wrong selectivity. Thus, one intermediate condition can be feasible joining both benefits of crude extract and pure enzymes. For that result, several procedures of extraction and semi purification of polyphenol oxidase (PPO) enzyme from banana (Musa sp.) were studied. The results showed that cleaned enzymatic extracts presented higher specific activities than crude extracts (2.8 up to 5.3 fold), despite the total protein concentration diminishing from 27% up to 72%, indicating that polyphenol oxidase (PPO) enzyme was preserved in them. The biosensors with 125 AU mL-1 containing cleaned enzymatic extracts performed better by grinding or grinding plus sonication for 30 s. They were linear over the ranges of 5.9 × 10-6 mol·L-1 to 1.4 × 10-3 mol·L-1 and 7.9 × 10-8 mol·L-1 to 4.0 × 10-3 mol·L-1, respectively. The limit of Detection (LOD) was 5.9 × 10-6 mol·L-1 and 7.9 × 10-8 mol·L-1, respectively. The LOD obtained is adequate to adrenaline determination on blood and medicinal samples, and were applied in medicinal samples with satisfactory results.

超声辅助酶法提取玉米胚芽粕水解蛋白及其抗氧化活性

以玉米胚芽粕为原料,研究超声波辅助碱性蛋白酶提取玉米胚芽粕水解蛋白的工艺条件。通过单因素试验初步确定超声波辅助酶法提取玉米胚芽粕水解蛋白的条件,在单因素试验基础上进行正交试验设计,并测定其抗氧化活性。结果表明,最优提取条件为超声温度50℃、超声时间50 min、超声功率140 W,在此条件下,玉米胚芽粕的水解蛋白提取率为90.6%。对DPPH自由基和羟自由基的IC_(50)分别为17.56µg/mL和28.41µg/mL。

熵权-正交法优化赤苍藤中多糖、多酚和黄酮的提取工艺

优化赤苍藤中多糖、多酚和黄酮的提取工艺,为综合评价其质量提供参考。以赤苍藤茎为研究对象,采用酶解法,选取适宜的酶质量分数、酶解p H、酶解温度及酶解时间为考察因素,以多糖、多酚、黄酮和浸膏提取率为评价指标。基于熵权法客观赋权结合L_9(3~4)正交试验,优选赤苍藤的多指标提取工艺。最佳提取工艺条件为:酶质量分数4%,酶解p H值6.0,酶解温度50℃,酶解时间45 min。考察多指标后,确认优选的提取工艺稳定可行,可为赤苍藤的质量评价提供实验依据。

山楂多糖酶解法和水提法的工艺优化和比较

研究酶解法和水提法对山楂多糖提取率的影响,为进一步开发利用山楂多糖提供技术依据。分别对酶解法和水提法进行优化,选择最佳条件提取山楂多糖。结果表明,酶解法提取山楂多糖的最适条件为酶用量3‰、酶解温度50℃、酶解时间1.0 h、酶解pH=8、料液比1∶10,多糖提取率为11.72%;水提法提取山楂多糖的最适条件为提取次数3次、提取时间120 min、料液比1∶10,多糖提取率为9.47%。酶解法提取率高,优于水提法。结论:山楂中多糖含量较高,通过酶解法可进行高效提取,方法简便,实用性强。

超声辅助水酶法提取花生油脂工艺研究及油脂品质分析

为了实现花生油脂的高效提取和利用,采用超声辅助水酶法提取花生油脂,研究超声功率、作用时间、作用温度、料液比及酶用量对花生油脂提取率的影响,采用响应面试验优化提取工艺,并分析花生油脂的脂肪酸组成、酸价、过氧化值和抗氧化能力。结果表明:花生油脂的最优提取工艺条件为料液比1∶5 g/mL、纤维素酶用量1.0%、超声功率247 W、作用时间41 min、作用温度51℃,此条件下的油脂提取率为92.64%±0.73%;从提取的花生油脂中共检出8种脂肪酸,其中油酸含量最高,为40.44%±0.57%,亚油酸含量为36.97%±0.05%;花生油脂的酸价为(0.38±0.04)mg/g,过氧化值为(0.15±0.00)g/100 g,DPPH和ABTS自由基清除率分别为64.82%±2.51%、82.56%±6.47%。超声辅助水酶法有效缩短了提取时间,降低了酶用量,并且制备的花生油脂中不饱和脂肪酸含量高,具有良好的抗氧化能力。