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.

Optimization of Enzymatic Extraction Process of Polysaccharides from Pseudostellaria heterophylla Fibrous Roots by Response Surface Methodology and Its Pilot Application

[Objectives]To study and optimize the process conditions of enzymatic hydrolysis technology for extracting polysaccharides from Pseudostellaria heterophylla fibrous roots and its application in workshop pilot tests.[Methods]P.heterophylla fibrous roots were taken as the matrix material,and Box Behnken design was used to analyze the extraction time,composite enzyme addition amount,and liquid-solid ratio for response surface optimization experiments,and then applied to the pilot extraction of P.heterophylla fibrous roots.[Results]Response surface analysis showed that all factors had a significant impact on the experimental indicators.The optimal extraction process conditions for polysaccharides from P.heterophylla fibrous roots were extraction time of 2.7 h,compound enzyme addition of 2.5%,and liquid-solid ratio of 32.The yield of polysaccharides from P.heterophylla fibrous roots was 4.83%.The water extraction process of P.heterophylla fibrous roots extraction pilot was used as the control group for response surface optimization of the pilot experiment.The optimization results showed that the extraction time was 3 h,the amount of composite enzyme added was 2.5%,and the liquid-solid ratio was 28.The polysaccharide yield was 4.75%,an increase of 4.63%compared to the control group.[Conclusions]This paper could provide feasibility for the innovation of enzy-matic hydrolysis technology for P.heterophylla fibrous roots and its workshop pilot practice application,as well as a reference for the industrial application of its medicinal resources.

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.

Ultrasound-assisted Enzymatic Extraction of SDF on Passion Fruit Seeds

Ultrasound-assisted enzymatic extraction of soluble dietary fiber(SDF) from passion fruit seeds was carried out in order to shorten the extraction time and increase the yield of SDF. The effects of cellulase concentration, ultrasonic power, extraction temperature, solid-liquid ratio and water bath time on the SDF yield of passion fruit were studied by single factor analysis. And the orthogonal experiment was designed to optimize the extraction conditions. The results showed that the optimum conditions for the extraction of SDF from passion fruit by ultrasoundassisted enzymatic method were cellulase concentration 1.5%, ultrasonic power 120 W, solid-liquid ratio(g/mL) 1 ∶ 15, extraction temperature 55 ℃, and water bath time 160 min. The SDF extraction rate was determined to be 5.12%.

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.

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%).

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.

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.

Comparative Evaluation of Peptidases Produced by Penicillium corylophilum and Penicillium waksmanii in Solid State Fermentation Using Agro-industrial Residues

In this present work, the best conditions for production of peptidases under solid state fermentation by the fungi Penicillium corylophilum and Penicillium waksmanii, partial purification using Sephadex G-75 gel filtration column, as well as the biochemical characterization of the partial purified enzymes were investigated. P. corylophilum showed the best production in medium containing wheat bran, agro-industrial residue, without additives （egg albumin or casein）, in which peptidase activity reached 520 U mL^-1 and the enzyme displayed the optimum activity between pH range from 7 to 8 and 60 ℃. It also showed high stability in wide pH range and temperature until 45 ℃ for 60 min of incubation. On the other hand, P. waksmanii, the best production was noted in a medium containing wheat bran （95%） and casein （5%）, reaching 545 U mL^-1, with proteolytic optimum activity at pH 7.5 and 55 ℃. The enzyme was mainly stable in pH range from 8 to 9 and at temperatures until 45 ℃ for 60 rain of incubation. The peptidases secreted by both fungi were inhibited in the presence of phenylmethane sulfonyl fluoride, showing that they belong to the subclass of serine peptidases.

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.

EXTRACTION AND EVALUATION OF EDIBLE OIL FROM SCHIZOCHYTRIUM SP.USING AN AQUEOUS ENZYMATIC METHOD

Schizochytrium sp.,a marine microalga,is a potential source of edible oil due to its short growth cycle and rapid lipid accumulation,especially of docosahexaenoic acid.An approach to isolate edible microalgal oil from Schizochytrium sp.using aqueous enzymatic extraction(AEE)was developed.Parameters were optimized by single-factor experiments followed by Box-Behnken design.Proteases were effective in extracting oil.The maximum free oil recovery(49.7%±0.58%)and total oil recovery(68.1%±0.94%)were obtained under optimum conditions of liquid-to-solid ratio of 4.8:1,a 2.5%enzyme concentration of papain and an extraction time of 2.2 h.There was a significant difference(P<0.05)in polyunsaturated fatty acid composition between microalgal oil obtained by AEE and by Soxhlet extraction,with the former having superior physiochemical properties and higher concentrations of bioactive components including total phenolic compounds and total tocopherols.These findings indicate a potential application of AEE for extraction of oil from Schizochytrium sp.