Optimized biotransformation of acid-treated water melon peel hydrolyzate into ethanol / Biotransformação otimizada de hidrolisado de casca de melancia tratada com ácido em etanol

Abstract Today, global focus of research is to explore the solution of energy crisis and environmental pollution. Like other agricultural countries, bulk quantities of watermelon peels (WMP) are disposed-off in environment as waste in Pakistan and appropriate management of this waste is the need of hour to save environment from pollution. The work emphasizes the role of ethanologenic yeasts to utilize significant sugars present in WMP for low-cost bioethanol fermentation. Dilute hydrochloric acid hydrolysis of WMP was carried out on optimized conditions employing RSM (response surface methodology) following central composite design (CCD). This experimental design is based on optimization of ethanologenesis involving some key independent parameters such as WMP hydrolysate and synthetic media ratio (X1), incubation temperature (X2) and incubation temperature (X3) for maximal ethanol yield exploiting standard (Saccharomyces cerevisiae K7) as well as experimental (Metchnikowia cibodasensisY34) yeasts. The results revealed that maximal ethanol yields obtained from S. cerevisiae K7 was 0.36±0.02 g/g of reducing sugars whereas M. cibodasensisY34, yielded 0.40±0.01 g ethanol/g of reducing sugars. The yeast isolate M. cibodasensisY34 appeared as promising ethanologen and embodies prospective potential for fermentative valorization of WMP-to-bioethanol.

Resumo Hoje, o foco global da pesquisa é explorar a solução da crise energética e da poluição ambiental. Como em outros países agrícolas, grandes quantidades de cascas de melancia (WMP) são descartadas como resíduos no meio ambiente no Paquistão, mas a gestão adequada desses resíduos é a mais recente solução para salvar o meio ambiente da poluição. O trabalho enfatiza o papel das leveduras etanologênicas para utilizar açúcares significativos presentes no WMP para fermentação de bioetanol de baixo custo. A hidrólise de ácido clorídrico diluído de WMP foi realizada em condições otimizadas empregando RSM (metodologia de superfície de resposta) e seguindo o projeto de composto central (CCD). Este projeto experimental é baseado na otimização da etanologenesis envolvendo alguns parâmetros independentes importantes, como hidrolisado de WMP e razão de meio sintético (X1), temperatura de incubação (X2) e temperatura de incubação (X3) para rendimento máximo de etanol explorando o padrão (Saccharomyces cerevisiae K7) também como leveduras experimentais (Metchnikowia cibodasensis Y34). Os resultados revelaram que os rendimentos máximos de etanol obtidos a partir de S. cerevisiae K7 foi de 0,36 ± 0,02 g / g de açúcares redutores, enquanto M. cibodasensis Y34 rendeu 0,40 ± 0,01 g de etanol / g de açúcares redutores. O isolado de levedura M. cibodasensis Y34 apareceu como um etanologeno promissor e incorpora um potencial prospectivo para a valorização fermentativa de WMP em bioetanol.

Optimized biotransformation of acid-treated water melon peel hydrolyzate into ethanol / Biotransformação otimizada de hidrolisado de casca de melancia tratada com ácido em etanol

Today, global focus of research is to explore the solution of energy crisis and environmental pollution. Like other agricultural countries, bulk quantities of watermelon peels (WMP) are disposed-off in environment as waste in Pakistan and appropriate management of this waste is the need of hour to save environment from pollution. The work emphasizes the role of ethanologenic yeasts to utilize significant sugars present in WMP for low-cost bioethanol fermentation. Dilute hydrochloric acid hydrolysis of WMP was carried out on optimized conditions employing RSM (response surface methodology) following central composite design (CCD). This experimental design is based on optimization of ethanologenesis involving some key independent parameters such as WMP hydrolysate and synthetic media ratio (X1), incubation temperature (X2) and incubation temperature (X3) for maximal ethanol yield exploiting standard (Saccharomyces cerevisiae K7) as well as experimental (Metchnikowia cibodasensisY34) yeasts. The results revealed that maximal ethanol yields obtained from S. cerevisiae K7 was 0.36±0.02 g/g of reducing sugars whereas M. cibodasensisY34, yielded 0.40±0.01 g ethanol/g of reducing sugars. The yeast isolate M. cibodasensisY34 appeared as promising ethanologen and embodies prospective potential for fermentative valorization of WMP-to-bioethanol.

Hoje, o foco global da pesquisa é explorar a solução da crise energética e da poluição ambiental. Como em outros países agrícolas, grandes quantidades de cascas de melancia (WMP) são descartadas como resíduos no meio ambiente no Paquistão, mas a gestão adequada desses resíduos é a mais recente solução para salvar o meio ambiente da poluição. O trabalho enfatiza o papel das leveduras etanologênicas para utilizar açúcares significativos presentes no WMP para fermentação de bioetanol de baixo custo. A hidrólise de ácido clorídrico diluído de WMP foi realizada em condições otimizadas empregando RSM (metodologia de superfície de resposta) e seguindo o projeto de composto central (CCD). Este projeto experimental é baseado na otimização da etanologenesis envolvendo alguns parâmetros independentes importantes, como hidrolisado de WMP e razão de meio sintético (X1), temperatura de incubação (X2) e temperatura de incubação (X3) para rendimento máximo de etanol explorando o padrão (Saccharomyces cerevisiae K7) também como leveduras experimentais (Metchnikowia cibodasensis Y34). Os resultados revelaram que os rendimentos máximos de etanol obtidos a partir de S. cerevisiae K7 foi de 0,36 ± 0,02 g / g de açúcares redutores, enquanto M. cibodasensis Y34 rendeu 0,40 ± 0,01 g de etanol / g de açúcares redutores. O isolado de levedura M. cibodasensis Y34 apareceu como um etanologeno promissor e incorpora um potencial prospectivo para a valorização fermentativa de WMP em bioetanol.

Optimized biotransformation of acid-treated water melon peel hydrolyzate into ethanol

Abstract Today, global focus of research is to explore the solution of energy crisis and environmental pollution. Like other agricultural countries, bulk quantities of watermelon peels (WMP) are disposed-off in environment as waste in Pakistan and appropriate management of this waste is the need of hour to save environment from pollution. The work emphasizes the role of ethanologenic yeasts to utilize significant sugars present in WMP for low-cost bioethanol fermentation. Dilute hydrochloric acid hydrolysis of WMP was carried out on optimized conditions employing RSM (response surface methodology) following central composite design (CCD). This experimental design is based on optimization of ethanologenesis involving some key independent parameters such as WMP hydrolysate and synthetic media ratio (X1), incubation temperature (X2) and incubation temperature (X3) for maximal ethanol yield exploiting standard (Saccharomyces cerevisiae K7) as well as experimental (Metchnikowia cibodasensisY34) yeasts. The results revealed that maximal ethanol yields obtained from S. cerevisiae K7 was 0.36±0.02 g/g of reducing sugars whereas M. cibodasensisY34, yielded 0.40±0.01 g ethanol/g of reducing sugars. The yeast isolate M. cibodasensisY34 appeared as promising ethanologen and embodies prospective potential for fermentative valorization of WMP-to-bioethanol.

Resumo Hoje, o foco global da pesquisa é explorar a solução da crise energética e da poluição ambiental. Como em outros países agrícolas, grandes quantidades de cascas de melancia (WMP) são descartadas como resíduos no meio ambiente no Paquistão, mas a gestão adequada desses resíduos é a mais recente solução para salvar o meio ambiente da poluição. O trabalho enfatiza o papel das leveduras etanologênicas para utilizar açúcares significativos presentes no WMP para fermentação de bioetanol de baixo custo. A hidrólise de ácido clorídrico diluído de WMP foi realizada em condições otimizadas empregando RSM (metodologia de superfície de resposta) e seguindo o projeto de composto central (CCD). Este projeto experimental é baseado na otimização da etanologenesis envolvendo alguns parâmetros independentes importantes, como hidrolisado de WMP e razão de meio sintético (X1), temperatura de incubação (X2) e temperatura de incubação (X3) para rendimento máximo de etanol explorando o padrão (Saccharomyces cerevisiae K7) também como leveduras experimentais (Metchnikowia cibodasensis Y34). Os resultados revelaram que os rendimentos máximos de etanol obtidos a partir de S. cerevisiae K7 foi de 0,36 ± 0,02 g / g de açúcares redutores, enquanto M. cibodasensis Y34 rendeu 0,40 ± 0,01 g de etanol / g de açúcares redutores. O isolado de levedura M. cibodasensis Y34 apareceu como um etanologeno promissor e incorpora um potencial prospectivo para a valorização fermentativa de WMP em bioetanol.

Optimization of extraction process of prescription medicinal materials of compound Yangshe granules / 药学实践杂志

Objective Optimizing the extraction process of prescription medicinal materials of hospital preparation of compound Yangshe granules. Methods A high performance liquid chromatograph (HPLC) quantitative method was established for deacetyl asperulosidicacid methyl ester (DME) and ferulic acid (FC) of the active ingredient. Based on the content of DME, FC and the yield of extract, the extraction process of compound Yangshe granule extract was optimized using central composite design-response surface methodology. Results The established HPLC method of quantification of active components in compound Yangshe granules met the requirements of method validation. The optimal extraction process optimized by central composite design-response surface methodology were as follows: the weight of extraction solvent was 12 times of the medicinal slices, the alcohol concentration was 73% and the extraction time was 60 min. Conclusion In this study, the quantitative method of active components in compound Yangshe granule by HPLC has been successfully established, and the optimized extraction process is simple and easy to operate with good repeatability.

Optimization of the extraction process of Shangke Huoxue Granule by central composite design-response surface methodology / 国际中医中药杂志

Objective:To optimize the extraction process of Shangke Huoxue Granule.Methods:Taking the factors of extraction solvent multiple, extraction time and extraction times as investigation factors, and extraction amount of ferulic acid, paeoniflorin and the ratio of extraction as comprehensive evaluation indices, one-factor experimental design and central composite design-response surface methodology were adopted to optimize the extraction process of Shangke Huoxue Granule.Results:The binomial fitting equation was Y=96.16+2.42 A+0.63 B-3.76 AB-1.57 A2-1.87 B2 ( P<0.01). The optimal extraction process parameters were confirmed to be adding 16 times of water, 64 minutes each time, twice. The deviation rates between the measured values of three verification experiments and the predicted value were 2.00%, 3.23% and 0.66%. Conclusion:The established model of central composite design-response surface methodology has high predictability and the optimized extraction process is stable and feasible.

Optimization of the preparation process of ethanol extracts from Centipeda minima and study on its anti-inflammatory activity of different extraction sites / 中国药房

OBJECTIVE To optimize the preparation technology of ethanol extracts from Centipeda minima， and investigate the anti-inflammatory activities of different extraction sites. METHODS Single factor test and response surface methodology were adopted to investigate the effects of ethanol volume fraction， extraction time， solid-liquid ratio and extraction times on the heating reflux extraction technology of total triterpenoids ethanol extract using the extraction rate of total triterpenoids of C. minima as indexes， optimize the extraction technology and carry out validation. Using dexamethasone as positive control drug， the effects of different extraction sites of C. minima （petroleum ether part， ethyl acetate part， n-butanol part， water part） on nitric oxide （NO） production in mononuclear macrophage RAW 264.7 cells of mice induced by lipopolysaccharide （LPS） were compared； the half inhibitory concentration （IC50） was calculated. RESULTS The optimal extraction technology of total triterpenoids ethanol extracts of C. minima was as follows： ethanol volume fraction of 70%， solid-liquid ratio of 1∶40 （g/mL）， extraction time of 2.0 h and extraction times of 3 times. The 3 times of validation tests showed that average extraction rate of total triterpenoids of C. minima was 1.134%， relative error of which with the predicted value was 0.02%. The petroleum ether part and ethyl acetate part of C. minima could inhibit the generation of NO in RAW 264.7 cells induced by lipopolysaccharide to different degrees. IC50 values of NO production were 2.44 μg/mL and 2.22 μg/mL， respectively， and both of them were lower than those of positive control drug dexamethasone （7.65 μg/mL）. CONCLUSIONS The optimized preparation process of ethanol extracts from C. minima is stability and feasibility. The petroleum ether part and ethyl acetate part have obvious anti-inflammatory effects.

Preparation of Huoluo Xiaoling gel plaster and its transdermal penetration in vitro / 中国中药杂志

Huoluo Xiaoling Dan is a classical prescription commonly used for blood circulation and pain relief in clinic with obvious effects. To make it directly treat lesion and improve the effect, this research optimized the preparation process of Huoluo Xiaoling gel paste and further evaluated its in vitro transdermal absorption performance, so as to provide a scientific basis for its development and utilization. Using primary viscosity, holding viscosity, and sensory score as evaluation indexes, the matrix amount of gel paste was determined by the single factor test and Box-Behnken response surface method. The ultra-performance liquid chromatography(UPLC) method was established to determine the content of eight active ingredients, including Danshensu, ferulic acid, salvianolic acid B, salvianolic acid A, ligustilide, tanshinone Ⅱ_A, 11-keto-β-boswellic(KBA), and 3-acetyl-11-keto-β-boswellic acid(AKBA). A mo-dified Franz diffusion cell method was used to evaluate and compare the absorption properties of the gel paste without volatile oil and with volatile oil microemulsion. The results showed that the optimal prescription for Huoluo Xiaoling gel paste matrix was NP700(1.35 g), glycerol(7.00 g), micropowder silica gel(1.25 g), sodium carboxymethyl cellulose(0.20 g), tartaric acid(0.06 g), and glyceryl aluminum(0.04 g). The mass fractions of eight active ingredients in the paste were successively 0.48, 0.014, 0.95, 0.39, 0.57, 0.055, 0.35, and 0.97 mg·g~(-1). The results of the in vitro transdermal absorption test showed that the addition of the volatile oil or the volatile oil microemulsion promoted the transdermal absorption of the active ingredients, and the law of drug penetration conformed to the zero equation or the Higuchi equation. The gel paste prepared by the optimal prescription has good appearance and adhesion, with no residue, and has the characteristics of skeletal slow-release preparation, which is easy to reduce the number of administration, la-ying a foundation for the development of new external dosage forms of Huoluo Xiaoling Dan.

Processing Magnoliae Officinalis Cortex with ginger juice: process optimization based on AHP-CRITIC weighting method and composition changes after processing / 中国中药杂志

The weight coefficients of appearance traits, extract yield of standard decoction, and total content of honokiol and magnolol were determined by analytic hierarchy process(AHP), criteria importance though intercrieria correlation(CRITIC), and AHP-CRITIC weighting method, and the comprehensive scores were calculated. The effects of ginger juice dosage, moistening time, proces-sing temperature, and processing time on the quality of Magnoliae Officinalis Cortex(MOC) were investigated, and Box-Behnken design was employed to optimize the process parameters. To reveal the processing mechanism, MOC, ginger juice-processed Magnoliae Officinalis Cortex(GMOC), and water-processed Magnoliae Officinalis Cortex(WMOC) were compared. The results showed that the weight coefficients of the appearance traits, extract yield of standard decoction, and total content of honokiol and magnolol determined by AHP-CRITIC weighting method were 0.134, 0.287, and 0.579, respectively. The optimal processing parameters of GMOC were ginger juice dosage of 8%, moistening time of 120 min, and processing at 100 ℃ for 7 min. The content of syringoside and magnolflorine in MOC decreased after processing, and the content of honokiol and magnolol followed the trend of GMOC>MOC>WMOC, which suggested that the change in clinical efficacy of MOC after processing was associated with the changes of chemical composition. The optimized processing technology is stable and feasible and provides references for the modern production and processing of MOC.

Trametes versicolor (L.) Lloyd as a source of thermostable serine protease: production and characterization

Proteases are ubiquitously present and are among the largest groups of commercially important enzymes. Here, we investigated a wood-rot basidiomycete Trametes versicolor (L.) Lloyd [Syn. Coriolus versicolor (L.) Quél.; Polyporus versicolor (L.) Fr.] as a source of the enzyme serine protease, its production, and optimized to obtain a higher yield of the enzyme.. The significant variables with optimized values for maximum production of the enzyme were temperature (30?C), incubation time (120 h) and wheat bran (10 g). The yield increased by 30.76% by statistically optimizing the media. The optimized temperature and pH for the maximum protease activity was 50?C and pH 7.0, respectively. The enzyme was purified through ion exchange (using DEAE cellulose 52 resin) and gel filtration chromatography (using Superdex 200 column). The purified enzyme had a retention time of 7 min in RP-HPLC. The enzyme was stable at a broad range of temperature (30-60?C) and pH (5.0-8.0) with a half-life of 58.72 min, Vmax of 37.17 ?M min/mL and Km of 0.657 mg/mL. Its activity was enhanced by Na+, Ca2+, Mg2+ ions and SDS surfactant. These properties make this enzyme a valuable candidate for industrial applications

Bioconversion of glycerol waste to ethanol by Escherichia coli and optimisation of process parameters

Biofuel is one of the best ways to reduce our dependence on fossil fuels. Ever since commercial biodiesel production began, waste glycerol, the biodiesel byproduct, has gained researchers’ interest, especially its recycling. Here, we explored using glycerol residue (carbon source) as a substrate in the fermentation process for ethanol production by Escherichia coli K12 in anaerobic conditions. The factors affecting the ethanol production was optimised by response surface methodology (RSM). Significant variables that impact the ethanol concentration were pH, temperature and the substrate, with a statistically significant effect (P <0.05) on ethanol formation. The significant factor was analyzed by the Box-Behnken design. The optimum conditions for bioethanol formation using glycerol as substrate was obtained at pH 7 and temperature 37°C. The ethanol productivity was 0.77 g/L/h. The ethanol concentration of 9.2 g/L achieved from glycerol residue was close to the theoretical value with the fermentation achieved at optimised terms.

High protein, gluten-free cookies made with rice, beans and avocado

Cookies are versatile foods that can supply specific needs and can be used as food vehicles to increase the intake of various nutrients. The objective was to create a gluten-free butter cookie based on rice flour (Oryza sativa). A central composite design 22 was used for analyzing the substitution of rice flour for bean flour (Phaseolus vulgaris, L.) (0-60%) and the substitution of butter for avocado puree (Persea americana) (0-100%). Response Surface Methodology were used to analyze the data with a significance of 10% (p<0.1) and a minimum R2 of 0.6. Variables analyzed were protein content (%), fat content (%), spread ratio, weight loss (%), water activity, moisture content (%), and ash content (%). Increases in the percentage of substitution of bean flour and avocado puree increased the amount of protein, ash, and moisture significantly. Fat content increased significantly following the decrease in avocado substitution. The minimum weight loss was obtained on intermediate values of bean flour. Water activity and spread ratio were not affected by changes in the variables measured. Three different formulas were obtained for the optimization: 46% bean flour and 86% of avocado puree, a formulation with an appropriate moisture range by reducing the lipid oxidation (59% bean flour and 82% avocado puree), and a formulation with 100% avocado (40% bean flour and 100% avocado puree). Sensory evaluation results of optimized treatments indicated that the formula with 46% bean flour and 86% avocado puree presented the highest global acceptance. Results from this study showcase the possibility of producing gluten-free cookies with good protein content.

Las galletas son alimentos versátiles que pueden suplir necesidades específicas y ser utilizados como vehículos para aumentar la ingesta de distintos nutrientes. El objetivo fue elaborar galletas de mantequilla libres de gluten a base de harina de arroz. Se realizó un diseño central compuesto 22 donde se estudió la sustitución parcial de harina de arroz por harina de frijol (Phaseolus vulgaris, L.) (0-60%) y la sustitución de la mantequilla por puré de aguacate (Persea americana) (0-100%). Los resultados fueron analizados por la Metodología de Superficie de Respuesta con un nivel de significancia del 10% (p<0.1) y con un mínimo R2 de 0.6. Las variables de respuesta fueron: proteína (%), grasa (%), diámetro/altura, pérdida de peso (%), humedad (%) y cenizas (%). A valores altos de las dos variables estudiadas la cantidad de proteína, de cenizas y de humedad aumentó significativamente. La cantidad de grasa se incrementó al utilizar bajos niveles de puré de aguacate. Los menores valores de pérdida de peso se obtuvieron en niveles intermedios de harina de frijol. La relación diámetro/altura no fue afectada por las variables estudiadas. Al optimizar se obtuvieron 3 formulaciones diferentes: 46% de harina de frijol y 86% puré de aguacate; 59% de frijol y 82% de puré de aguacate y otra (no estadística) 40% de frijol y 100% de puré de aguacate. Los resultados del análisis sensorial de las muestras optimizadas indicaron que la formulación con mayor aceptación global fue la elaborada con 46% de harina de frijol y 86% de puré de aguacate, concluyendo que es posible elaborar una galleta libre de gluten y con una adecuada cantidad de proteína.

Decolorization of anthraquinone-based dye (Vat Brown R) by Pseudomonas aeruginosa NCH - Optimization and kinetic study

The objective of this research work is related to the fact that the source of isolation and acclimatization process influences the microorganism’s potential for the decolorization of various substances. Some of the widely used anthraquinone vat dyes decolorization by the pure bacterial strain is a significant aspect that will assist in the in-situ bioremediation of the ecosystem.The present study is to evaluate the enhanced decolorization of Vat Brown R by an isolated bacterium, Pseudomonas aeruginosa NCH, from textile dye wastewater under aerobic conditions. The effect of pH, temperature, and inoculum size was optimized using response surface methodology with the box-behnken experimental design. The strain NCH showed maximum decolorization efficacy under optimum conditions at pH9.76, temperature 34.69?C, and an inoculum size of 9.51% (v/v), respectively. A decolorization of 90.34% was observed with 100 mg L?1of Vat Brown R within 18h under these conditions. Confirmatory experiments have verified the optimum combination of the three variables predicted by RSM. Kinetics study was carried out using various approaches: Michaelis-Menten (Vmax = 29.1 mg L?1 h?1 and Km = 25.2 mg L?1), Lineweaver-Burk (Vmax = 30.12 mg L?1 h?1 and Km = 26.91 mg L?1), and Eadie-hofstee model (Vmax = 30.23 mg L?1h?1and Km = 27.29 mg L?1), and the results showed that the degradation followed a first-order reaction kinetics. The subsequent degradation of the dye and the formation of metabolites were studied using analytical techniques such as UV-vis spectroscopy and FT-IR analysis. UV-vis spectroscopy validated the detoxification of the dye and confirmed that Pseudomonas sp. NCH overcomes this decolorizing activity through biodegradation. This study investigated the highest decolorization efficiency of strain NCHused in the biodegradation of wastewaters containing anthraquinone dyes.

Optimization of preparation technology of baicalin lipid nano foam aerosol / 中国药房

OBJECTIVE To optimize the pr eparation technology of the baicalin lipid nano foam aerosol （BC-LN-FA）. METHODS Baicalin lipid nanoparticle （BC-LN）and BC-LN-FA were prepared by the thin film dispersion method and homogeneous emulsification method ，respectively，using baicalin （BC） as the model drug. The preparation technology was optimized by Box-Behnken design-response surface methodology using particle size and encapsulation efficiency （EE）as indexes ，with dosage ， emulsifier dosage ，co-emulsifier dosage and homogenization time as factors. The morphology ，particle size ，polymerdispersity index（PDI），EE，the viscosity ，the foam dissolution rate and in vitro transdermal release of BC-LN-FA were characterized. RESULTS The optimal technology included 25 mg BC ，40 mg emulsifier （mass ratio of stearic acid-soybean lecithin-glycerol was 1∶1∶1），30 mg co-emulsifier （mass ratio of octadecanol-lactic acid was 1∶1），homogenization time of 20 min. Results of 3 times of validation tests showed that particle size of prepared BC-LN-FA was （151.70±2.40）nm，EE was （68.62±1.16）％；the deviation of them from the predicted value （particle size of 150.80 nm，EE of 67.02％）were 0.60％ and 2.39％ respectively. The BC-LN-FA prepared by the optimal process was light yellow opalescence ，uniform in particle size and round-like in shape. The viscosity，the foam dissolution rate ，the content of BC and PDI were （122.92±5.09）mPa·s，（65.32±3.22）％，（7.01±0.12）％ and（0.199±0.006），respectively. At 48 h，the cumulative release rates of BC-LN-FA in phosphate buffer saline （PBS）at pH 7.4， 6.8，5.0 were（54.12±2.69）％，（57.85±4.25）％ and（59.47±1.83）％，respectively；those of free BC in PBS at pH 7.4 was only （15.04±1.43）％. CONCLUSIONS The optimized technology is stable and feasible. Prepared BC-LN-FA has a uniform particle size，high digestion rate and certain viscosity.

Acetylation of Rehmannia glutinosa polysaccharides and antioxidant activity of acetylated derivatives / 中国中药杂志

This study aims to acetylate Rehmannia glutinosa polysaccharides by acetic anhydride method, optimize process parameters and evaluate their antioxidant activity. With the degree of substitution(D_s) as a criterion, the effects of reaction time, acetic anhydride-to-polysaccharides ratio and temperature were investigated. Process parameters were optimized by single-factor experiment and response surface methodology. The infrared spectroscopy(IR) and scanning electron microscopy(SEM) proved the successful acetylation and were employed to preliminarily analyze the structural characteristics of acetylated derivatives. The results showed that the D_s was 0.327 under the optimal technological conditions, including m(acetic anhydride):m(R. glutinosa polysaccharides)=2.70, reaction time 3.0 h and temperature 48 ℃. Further, the antioxidant properties of acetylated derivatives were investigated in vitro and acetylation was found effective to improve the antioxidant activity of R. glutinosa polysaccharides. This study provides a reference for the further development and application of R. glutinosa polysaccharides.

Formulation optimization of emodin nanostructured lipid carriers by Box-Behnken response surface method and in vitro quality evaluation / 中国中药杂志

Emodin nanostructured lipid carriers(ED-NLC) were prepared and their quality was evaluated in vitro. Based on the results of single-factor experiments, the ED-NLC formulation was optimized by Box-Behnken response surface method with the dosages of emodin, isopropyl myristate and poloxamer 188 as factors and the nanoparticle size, encapsulation efficiency and drug loading as evaluation indexes. Then the evaluation was performed on the morphology, size and in vitro release of the nanoparticles prepared by emulsification-ultrasonic dispersion method in line with the optimal formulation, i.e., 3.27 mg emodin, 148.68 mg isopropyl myristate and 173.48 mg poloxamer 188. Under a transmission electron microscope(TEM), ED-NLC were spherical and their particle size distribution was uniform. The particle size of ED-NLC was(97.02±1.55) nm, the polymer dispersion index 0.21±0.01, the zeta potential(-38.96±0.65) mV, the encapsulation efficiency 90.41%±0.56% and the drug loading 1.55%±0.01%. The results of differential scanning calorimeter(DSC) indicated that emodin may be encapsulated into the nanostructured lipid carriers in molecular or amorphous form. In vitro drug release had obvious characteristics of slow release, which accorded with the first-order drug release equation. The fitting model of Box-Behnken response surface methodology was proved accurate and reliable. The optimal formulation-based ED-NLC featured concentrated particle size distribution and high encapsulation efficiency, which laid a foundation for the follow-up study of ED-NLC in vivo.

Optimization of molding process of Jinpuju Qingrelishi Granules by Response Surface Methodology / 国际中医中药杂志

Objective:To optimize the best molding process of Jinpuju Qingrelishi Granules.Methods:Based on the single factor test, the relative density of clear ointment and the amount of diluent (dextrin∶lactose=2∶1) are used as investigating factors, and the overall evaluation of the molding rate and angle of repose overall desirability (OD) is used as the evaluation index. The effect surface method is used to optimize the best molding process of Jinpuju Qingrelishi Granules.Results:The best molding process conditions: the relative density of the clear paste is 1.20 (60 ℃) and the amount of diluent is 3 times that of the clear paste. After mixing the clear paste and diluent, make soft material, pass through a 14-mesh sieve to granulate, dry in an oven (55 ℃) for 1 hour, and sizing to obtain. The molding rates of the three batches of verification test granules were 93.73%, 93.03%, 95.59%, respectively, the predicted OD value was 0.928, the verification value was 0.936, and the deviation from the predicted value was -0.86%.Conclusion:The molding process of this experiment is stable and reliable, with good repeatability, which can provide a reference for the follow-up research of Jinpuju Qingrelishi Granules.

Optimization of glycerol-based medium composition for antifungal metabolites production by Bacillus subtilis

Abstract Bacillus species are promising producers of various compounds that have pronounced antimicrobial, antiviral and antitumor activities. Due to its GRAS status, Bacillus subtilis represents an excellent candidate for the usage in plant pathogens biocontrol. In this research, evaluation of antifungal metabolites biosynthesis by Bacillus subtilis ATCC 6633 and optimization of glycerol-based medium composition, using response surface methodology, for the production of compounds effective against Neurospora crassa were investigated. The results of disc-diffusion method indicate that applied Bacillus strain produces compounds with antifungal activity against tested fungus. In order to find optimal cultivation medium composition, the experiments were carried out in accordance with Box-Behnken design, and optimization was performed using the concept of desirability function combined with previously defined mathematical equation, which describes examined bioprocess. The optimization model predicts that maximum inhibition zone diameter against Neurospora crassa of 32.24 mm is achieved when initial content of glycerol, NaNO2 and K2HPO4 were 49.68 g/L, 2.90 g/L and 6.49 g/L, respectively. Additionally, the second optimization set was made to minimize the consumption of medium components and costs of medium preparation. The obtained results are the basis for further research aimed to develop medium appropriate for economically justified production of bioactive compounds at industrial scale.

Optimization of ozone concentration and storage time in green asparagus (Asparagus officinalis L. ) using response surface methodology / Optimización de concentración de ozono y tiempo de almacenamiento en espárrago (Asparagus officinalis L. ) verde utilizando metodología de superficie de respuesta

Background: Asparagus (Asparagus officinalis L.) green is a vegetable with a great demand worldwide, and likewise, it is highly perishable, due to its high respiration rate that accelerates its senescence. Disinfection of vegetables after their harvest is an obligatory practice that can reduce losses by decomposition due to the attack of microorganisms. Therefore, it is vital to preserving its microbiological and sensory characteristics to reach the final consumer. Objective: to evaluate the effect of gaseous ozone (0 to 10 ppm) and storage time (0 to 30 days) on phenol content, overall appearance, count of molds, psychrophilic bacteria, and viable mesophilic aerobes. Methods: the response surface methodology was used, applying a rotatable central composite design. Results: the results indicated that there was a significant influence (p <0.05) of the independent variables on the characteristics studied, as well as an adequate lack of fit of the quadratic regression model (p> 0.05). By means of the contour superposition technique, it was determined that the optimal conditions for the highest retention of phenol content (16.99 mg/g) and overall appearance (7.61 points) and lower counts of viable aerobic mesophilic bacteria (5.3 x 103 CFU/g) they corresponded to 10 ppm of gaseous ozone up to 25.91 days of storage, with adequate quality characteristics in the spears. Conclusion: the region of interest was determined for optimal retention of phenol content and overall appearance, and a lower count of viable aerobic mesophilic bacteria in green asparagus during postharvest, suggesting to use the initial application of ozone gas at 10 ppm allowing 25.9 days storage at 1 °C. The results indicate that this technology is a good alternative in the conservation of fresh vegetables

Antecedentes: El espárrago (Asparagus officinalis L.) verde; es una hortaliza con una gran demanda a nivel mundial, y, asimismo, es altamente perecible, por su elevada velocidad de respiración que, acelera su proceso de senescencia. La desinfección de los vegetales después de su cosecha es una práctica obligada que puede disminuir las pérdidas por descomposición debido al ataque de microrganismos. Por lo tanto, es muy importante conservar sus características microbiológicas y sensoriales para llegar al consumidor final. Objetivo: evaluar el efecto del ozono gaseoso (0 a 10 ppm) y tiempo de almacenamiento (0 a 30 días) sobre el contenido de fenoles, apariencia general, recuento de mohos, bacterias psicrófilas y aerobias mesófilas viables. Métodos: se utilizó la metodología de superficie de respuesta, aplicando un diseño compuesto central rotable. Resultados: los resultados indicaron que existió influencia significativa (p<0.05) de las variables independientes sobre las características estudiadas, así como, una adecuada bondad de ajuste del modelo de regresión cuadrático (p>0.05). Mediante la técnica de superposición de contornos se determinó que las condiciones óptimas para la mayor retención de contenido de fenoles (16.99 mg/g) y apariencia general (7.61 puntos) y menor recuentos de bacterias aerobias mesófilas viables (5.3 x 103 UFC/g) correspondieron a 10 ppm de ozono gaseoso hasta los 25.91 días de almacenamiento, con adecuadas características de calidad en los turiones. Conclusión: se determinó la región de interés para una óptima retención de contenido de fenoles y apariencia general, así como, menor recuento de bacterias aerobias mesófilas viables en el espárrago verde durante la postcosecha, sugiriendo utilizar la aplicación inicial de ozono gaseoso a 10 ppm permitiendo 25.9 días de almacenamiento a 1 °C. Los resultados indican que esta tecnología es una buena alternativa en la conservación de hortalizas frescas

Statistical optimization of cellulases by Talaromyces thermophilus utilizing Saccharum spontaneum, a novel substrate

BACKGROUND: At present, cellulases are the most important enzymes worldwide, and their demand has been increasing in the industrial sector owing to their notable hydrolysis capability. RESULTS: In the present study, contrary to conventional techniques, three physical parameters were statistically optimized for the production of cellulase by thermophilic fungi by using response surface methodology (RSM). Among all the tested thermophilic strains, the best cellulase producing fungus was identified as Talaromyces thermophilus ­ both morphologically and molecularly through 5.8S/ITS rDNA sequencing. The central composite design (CCD) was used to evaluate the interactive effect of the significant factors. The CCD was applied by considering incubation period, pH, and temperature as the model factors for the present investigation. A second-order quadratic model and response surface method revealed that the independent variables including pH 6, temperature 50 C, and incubation period 72 h significantly influenced the production of cellulases. The analysis of variance (ANOVA) indicated that the established model was significant (P 0.05) and showed the high adequacy of the model. The actual and predicted values of CMCase and FPase activity showed good agreement with each other and also confirmed the validity of the designed model. CONCLUSIONS: We believe the present findings to be the first report on cellulase production by exploiting Kans grass (Saccharum spontaneum) as a substrate through response surface methodology by using thermophilic fungus, Talaromyces thermophilus.

Optimization of polysaccharide extraction from Hippocampus by deep neural network and Box-Behnken design-response surface methodology / 中国中药杂志

In this paper, the extraction rate of crude polysaccharides and the yield of polysaccharides from Hippocampus served as test indicators. The comprehensive evaluation indicators were assigned by the R language combined with the entropy weight method. The Box-Behnken design-response surface methodology(BBD-RSM) and the deep neural network(DNN) were employed to screen the optimal parameters for the polysaccharide extraction from Hippocampus. These two modeling methods were compared and verified experimentally for the process optimization. This study provides a reference for the industrialization of effective component extraction from Chinese medicinals and achieves the effective combination of modern technology and traditional Chinese medicine.