Response surface methodology as a tool to study the lipase‐catalyzed synthesis of betulinic acid ester

BACKGROUND: The synthesis of betulinic acid ester using betulinic acid and oleyl alcohol catalyzed by Novozym 435 (immobilized Candida antarctica lipase) was carried out. Response surface methodology (RSM) based on a five‐level, three‐variable, central composite rotatable design (CCRD) was employed to evaluate the interactive effects of various parameters. The parameters were reaction time (8–16 h), temperature (20–60 °C) and enzyme amount (120–160 mg). RESULTS: Simultaneously increasing reaction time, temperature and amount of enzyme increased the yields of betulinic acid ester produced. CONCLUSION: The optimum conditions derived via RSM for the reaction were reaction time of 10.2 h, temperature of 53.1 °C and enzyme amount of 138 mg. The actual experimental yield was 48.5% under optimum conditions, which compared well with the maximum predicted value of 47.6%. Copyright © 2008 Society of Chemical Industry     

Chemical and enzymatic interesterification of a beef tallow and rapeseed oil equal‐weight blend

A mixture of beef tallow and rapeseed oil (1:1, wt/wt) was interesterified using sodium methoxide or immobilized lipases from Rhizomucor miehei (Lipozyme IM) and Candida antarctica (Novozym 435) as catalysts. Chemical interesterifications were carried out at 60 and 90 °C for 0.5 and 1.5 h using 0.4, 0.6 and 1.0 wt‐% CH₃ONa. Enzymatic interesterifications were carried out at 60 °C for 8 h with Lipozyme IM or at 80 °C for 4 h with Novozym 435. The biocatalyst doses were kept constant (8 wt‐%), but the water content was varied from 2 to 10 wt‐%. The starting mixture and the interesterified products were separated by column chromatography into a pure triacylglycerol fraction and a nontriacylglycerol fraction, which contained free fatty acids, mono‐, and diacylglycerols. It was found that the concentration of free fatty acids and partial acylglycerols increased after interesterification. The slip melting points and solid fat contents of the triacylglycerol fractions isolated from interesterified fats were lower compared with the nonesterified blends. The sn‐2 and sn‐1,3 distribution of fatty acids in the TAG fractions before and after interesterification were determined. These distributions were random after chemical interesterification and near random when Novozym 435 was used. When Lipozyme IM was used, the fatty acid composition at the sn‐2 position remained practically unchanged, compared with the starting blend. The interesterified fats and isolated triacylglycerols had reduced oxidative stabilities, as assessed by Rancimat induction times. Addition of 0.02% BHA and BHT to the interesterified fats improved their stabilities.     

Biodiesel production from Nannochloropsis microalgal biomass-derived oil: An experimental and theoretical study using the RSM-CCD approach

Biodiesel production from Nannochloropsis is investigated in the current study. Based on the extraction studies, the used biomass is composed of 50% saponifiable lipids, which turns this species to a vulnerable feedstock for biodiesel production. It should also be noted that the acidity of the obtained crude bio-oil is higher than 2 mg KOH/g, hence it was attempted to survey the biodiesel production from the obtained oil through the esterification reaction with the aid of response surface methodology (RSM). According to the data, the highest biodiesel yield (99.5%) from the bio-oil is obtained at a reaction temperature of 69°C, a reaction time of 30 min, a methanol to oil molar ratio of 9:1, and an H₂SO₄ concentration of 0.13 wt.%. In the next step, the in-situ extraction and esterification of Nannochloropsis were investigated at the observed optimum reaction conditions. Based on the analysis, biodiesel yield from the dry cell weight of the biomass is considered to be 16%, which accounts for 99% conversion of bio-oil to biodiesel.     

Optimized reuse and bioconversion from retentate of pre‐filtered palm oil mill effluent (POME) into microbial protease by Aspergillus terreus using response surface methodology

BACKGROUND: The membrane filtration process enables the treatment of wastewater, producing permeate which is less polluted. However, disposal is usually required for the retentate, which is produced as a concentrated constituent along with the permeate. In this study, the authors explored the possibility of reusing, rather than disposing of, the retentate of pre‐filtered palm oil mill effluent (POME) as a fermentation substrate in protease production by a wild type strain of Aspergillus terreus IMI 282743. In addition, the quantitative and interactive effects of the concentration factor for retentate, temperature, inoculum concentration, and fermentation time on the optimization of protease production were investigated using response surface methodology (RSM). RESULTS: Using RSM, the optimum conditions were found to be a concentration factor of 7.27, temperature of 37.95 °C, inoculum concentration of 1.30% (v/v) and fermentation time of 3.83 days. The protease production was increased 4.37‐fold in comparison with the results obtained under non‐optimized conditions. CONCLUSION: To a certain extent, protease production could be enhanced with an increase in concentration factor and temperature, and a decrease of inoculum concentration and fermentation time. Also, POME retentate was found to be a good substrate for protease production with high product activity and without nutrient supplementation. Copyright © 2009 Society of Chemical Industry     

Optimized synthesis of lipase‐catalyzed biodiesel by Novozym 435

The ability of immobilized lipase from Candida antarctica (Novozym 435) to catalyze the alcoholysis of canola oil and methanol was investigated. Response surface methodology (RSM) and five–level–five–factor central composite rotatable design (CCRD) were employed to evaluate the effects of synthesis parameters, such as reaction time, temperature, enzyme concentration, substrate molar ratio of methanol to canola oil, and added water content on percentage weight conversion of canola oil methyl ester by alcoholysis. Reaction temperature and enzyme concentration were the most important variables. High temperature and superabundant methanol inhibited the ability of Novozym 435 to catalyze the synthesis of biodiesel. Based on the analysis of ridge max, the optimum synthesis conditions were as follows: reaction time 12.4 h, temperature 38.0 °C, enzyme concentration 42.3%, substrate molar ratio 3.5:1, and added water 7.2%. The predicted value was 99.4% weight conversion, and the actual experimental value was 97.9% weight conversion. Copyright © 2004 Society of Chemical Industry     

Modeling and optimization of lipase‐catalyzed synthesis of dilauryl adipate ester by response surface methodology

BACKGROUND: Adipate esters are used as low‐temperature and low‐viscosity plasticizers for polyvinyl chloride and its copolymers. In this work, optimization of lipase‐catalyzed production of dilauryl adipate was carried out using response surface methodology (RSM) based on a four‐factor‐five‐level central composite rotatable design (CCRD). Immobilized lipase from Candida antarctica (Novozym 435) was used as catalyst in this reaction. Various reaction parameters affecting the synthesis of adipate ester, including alcohol/acid molar ratio, amount of enzyme, temperature and reaction time, were investigated. RESULTS: Statistical analysis showed that the amount of enzyme was less significant than the other three factors. The optimal conditions for the enzymatic reaction were obtained at 5.7:1 substrate molar ratio using 0.18 g of enzyme at 53.1 °C for 282.2 min. Under these conditions the esterification percentage was 96.0%. CONCLUSIONS: The results demonstrated that response surface methodology can be applied effectively to optimize the lipase‐catalyzed synthesis of adipate ester. The optimum conditions can obtained be used to scale up the process. Copyright © 2008 Society of Chemical Industry     

A continuous ultrasound‐assisted packed‐bed bioreactor for the lipase‐catalyzed synthesis of caffeic acid phenethyl ester

BACKGROUND: The focus of this paper is the ultrasound‐assisted synthesis of caffeic acid phenethyl ester (CAPE) from caffeic acid and phenyl ethanol in a continuous packed‐bed bioreactor. Immobilized Novozym^® 435 (from Candida antarctica) is used as the catalyst. A three‐level–three‐factor Box–Behnken design and a response surface methodology (RSM) are employed to evaluate the effects of temperature, flow rate, and ultrasonic power on the percentage molar conversion of CAPE. RESULTS: Based on ridge max analysis, it is concluded that the optimum condition for synthesis is reaction temperature 72.66 °C, flow rate 0.046 mL min^?1, and ultrasonic power 1.64 W cm^?2. The expected molar conversion value is 97.84%. An experiment performed under these optimal conditions resulted in a molar conversion of 92.11 ± 0.75%. The enzyme in the bioreactor was found to be stable for at least 6 days. CONCLUSIONS: The lipase‐catalyzed synthesis of CAPE by an ultrasound‐assisted packed‐bed bioreactor uses mild reaction conditions. Enzymatic synthesis of CAPE is suitable for use in the nutraceutical and food production industries. Copyright © 2011 Society of Chemical Industry     

High efficiency bioethanol production from OPEFB using pilot pretreatment reactor

BACKGROUND: Current ethanol production processes using crops such as corn and sugar cane are well established. However, the utilization of cheaper biomasses such as lignocellulose could make bioethanol more competitive with fossil fuels while avoiding the ethical concerns associated with using potential food resources. RESULTS: Oil palm empty fruit bunches (OPEFB), a lignocellulosic biomass, was pretreated using NaOH to produce bioethanol. The pretreatment and enzymatic hydrolysis conditions were evaluated by response surface methodology (RSM). The optimal conditions were found to be 127.64 °C, 22.08 min, and 2.89 mol L^?1 for temperature, reaction time, and NaOH concentration, respectively. Regarding enzymatic digestibility, 50 FPU g^?1 cellulose of cellulase was selected as the test concentration, resulting in a total glucose conversion rate (TGCR) of 86.37% using the Changhae Ethanol Multi Explosion (CHEMEX) facility. Fermentation of pretreated OPEFB using Saccharomyces cerevisiae resulted in an ethanol concentration of 48.54 g L^?1 at 20% (w/v) pretreated biomass loading, along with simultaneous saccharification and fermentation (SSF) processes. Overall, 410.48 g of ethanol were produced from 3 kg of raw OPEFB in a single run, using the CHEMEX_50 L reactor. CONCLUSION: The results presented here constitute a significant contribution to the production of bioethanol from OPEFB. Copyright © 2011 Society of Chemical Industry     

Use optimization of natural antioxidants in refined, bleached, and deodorized palm olein during repeated deep-fat frying using response surface methodology

An optimization study on the use of oleoresin rosemary extract, sage extract, and citric acid added into refined, bleached, and deodorized (RBD) palm olein in deep-fat frying of potato chips was carried out using response surface methodology (RSM). Results showed that oleoresin rosemary extract was the most important factor affecting the sensory acceptability of potato chips. For taste and odor, its effects were highly significant (P<0.01), while for crispiness and overall acceptability, the effects were significant (P<0.05). As for sage extract, the level of this antioxidant had a highly significant (P<0.01) effect on appearance and taste and a significant effect (P<0.05) on odor and overall acceptability, but had no effect on crispiness. Although there was no significant synergistic correlation between citric acid and oleoresin rosemary extract or sage extract at the first order, its second order was significantly (P<0.05) related to taste, crispiness, and overall acceptability. An interaction between oleoresin rosemary and sage extracts also significantly (P<0.05) improved the score of overall acceptability of the potato chips. Contour maps of the sensory scores of potato chips indicated that the optimal points for appearance were achieved using 0.062% oleoresin rosemary extract, 0.066% sage extract, and 0.023% citric acid, while optimal task was achieved with 0.063% oleoresin rosemary extract, 0.075% sage extract, and 0.025% citric acid. With the same sequence of ingredients added into oil, the combinations required to achieve the optimal odor, crispiness, and overall acceptability scores were 0.058-0.046-0.026, 0.060-0.071-0.022, and 0.060-0.064-0.026%, respectively.     

Optimization of acrylonitrile removal by activated carbon‐granular using response surface methodology

A statistical Box–Behnken design of experiments was performed to evaluate the effects of individual operating variables and their interactions on the acrylonitrile (AN) removal of C₀ = 100 mg/L as fixed input parameter. The variables examined in this study included activated carbon‐granular (AC) dosage, w, temperature, T, and time of contact, t. The significant variables and optimum conditions were identified (w = 4 g/L, T = 30°C, and t = 120 min with AN uptake of 23.97 mg/g of AC) from statistical analysis of the experimental results using response surface methodology (RSM).     

Conjugated linoleic acid (CLA) production and lipase‐catalyzed interesterification of purified CLA with canola oil

In this study, two important isomers of CLA, i.e. c9,t11 and t10,c12, were produced up to ca. 73% of total fatty acids, employing alkali isomerization of safflower oil, followed by purification with only one‐step urea crystallization to 85.6%, while the recovery of the purification process was 35%. Interesterification (acidolysis) of purified CLA with canola oil was then conducted by Thermomyces lanuginosus lipase. The CLA content incorporated into the triacylglycerols (TG) was 26.6 mol‐% after 48 h of reaction time. Physical and chemical properties of the TG were then changed according to the degree of substitution of oleic acid in canola oil with CLA.     

Response Surface Optimization of the Critical Media Components for the Production of Surfactin

Optimization of the fermentation media for maximization of surfactin production was carried out. The carbon source (glucose), the nitrogen source (ammonium nitrate) and the mineral salts ferrous and manganous sulphates were the critical components of the medium optimized. A 2⁴ full factorial central composite experimental design followed by multi-stage Monte-Carlo optimization was used in the design of experiments and in the analysis of results. This procedure limited the number of actual experiments performed while allowing for possible interactions between the four components. The optimum values for the tested variables for the maximal production of surfactin were (in g dm⁻³): glucose = 36·5; NH₄NO₃ = 4·5; FeSO₄ = 4×10⁻³ and MnSO₄ = 27·5 ×10⁻². Relative surfactant concentrations were expressed as the reciprocal of the critical micelle concentration (CMC⁻¹) and the maximum predicted yield of surfactin in terms of CMC⁻¹ was 45·5. © 1997 SCI.     

响应面法优化麦芽糖水解制备5-羟甲基糠醛

以麦芽糖为原料,水为溶剂,考察了磷酸、硫酸、乙酸锌和氢氧化钠为催化剂对5-羟甲基糠醛制备的催化效果。研究结果表明：氢氧化钠的催化能力远高于其他催化剂,且用量较少。以5-羟甲基糠醛的收率为衡量标准,采取响应面法对反应条件进行了优化,得到最佳条件为反应温度95.20 ℃、催化剂氢氧化钠的质量2.25 g、反应时间6.17 h。在此反应条件下,5-羟甲基糠醛的收率达61.13 %。红外谱图分析结果与标准样品谱图一致,表明由麦芽糖成功制备了5-HMF。     

Lipase‐catalyzed ethanolysis of soybean oil in a solvent‐free system using central composite design and response surface methodology

BACKGROUND: In this work we describe the synthesis of ethyl esters, commonly known as biodiesel, using refined soybean oil and ethanol in a solvent‐free system catalyzed by lipase from Thermomyces lanuginosus. Central composite design and response surface methodology (RSM) were employed to optimize the biodiesel synthesis parameters, which were: reaction time, temperature, substrate molar ratio, enzyme content, and added water, measured as percentage of yield conversion. RESULTS: The optimal conditions obtained were: temperature, 31.5 °C; reaction time, 7 h; substrate molar ratio, 7.5:1 ethanol:soybean oil; enzyme content, 15% (g enzyme g⁻¹ oil); added water, 4% (g water g⁻¹ oil). The experimental yield conversion obtained under these conditions was 96%, which is very close to the maximum predicted value of 94.4%. The reaction time‐course at the optimal values indicated that 5 h was necessary to obtain high yield conversions. CONCLUSION: A high yield conversion was obtained under the optimized conditions, with relative low enzyme content and short time. Comparison of predicted and experimental values showed good correspondence, implying that the empirical model derived from RSM can be used to adequately describe the relationship between the reaction parameters and the response (yield conversion) in lipase‐catalyzed biodiesel synthesis. Copyright © 2008 Society of Chemical Industry     

Optimization of biodiesel production from crude palm oil using ultrasonic irradiation assistance and response surface methodology

BACKGROUND: Production of biodiesel from crude palm oil (CPO) with 6 wt% of free fatty acid (FFA) using a low‐frequency ultrasonic irradiation (40 kHz) technique was investigated in the present work. The objective of this study was to determine the relationship between various important parameters of the alkaline catalyzed transesterification process to obtain a high conversion to biodiesel. Response surface methodology (RSM) was used to statistically analyze and optimize the operating parameters of the process. A central composite design (CCD) was adopted to study the effects of the methanol to oil molar ratio, the catalyst concentration, reaction temperature, and irradiation time on conversion to biodiesel. RESULTS: The result from the RSM analysis indicated that the methanol to oil molar ratio, catalyst concentration and irradiation time have the most significant effects on the conversion to biodiesel. Moreover, a coefficient of determination (R²) value of 0.93 shows the fitness of a second‐order model for the present study. Based on this second‐order model, the optimum conditions for alkaline catalyzed transesterification of CPO were found to be a methanol to oil molar ratio of 6.44:1, catalyst concentration 1.25 wt%, reaction temperature 38.44 °C and irradiation time 25.96 min. At the calculated optimum condition, the conversion to biodiesel reached 97.85%. Under these same conditions, the experimental value was 98.02 ± 0.6%. CONCLUSIONS: The mathematical model developed has been proven to adequately describe the range of the experimental parameters studied and provide a statistically accurate prediction of the optimum conversion to biodiesel. Copyright © 2011 Society of Chemical Industry     

Multi response optimisation of polyphenol extraction conditions from grape seeds by using ultrasound assisted extraction (UAE)

Multi response optimisation conditions were investigated in grape seeds’ phenolic compounds extraction by using ultrasound-assisted extraction (UAE) methodology. The effect of independent process variables such as EtOH concentration (0–100%), extraction time (0–40 min), solvent:solid ratio (4.5–38.5 mL/g) and extraction temperature (20–60°C) on total phenolic content (TPC) and total antioxidant activity (TAA) of the extracts were studied. The optimum conditions of UAE were determined as follows: EtOH concentration, 61.76%; extraction time 20 min., solvent:solid ratio, 30 mL/g; extraction temperature 50°C. The estimation results of the model and the experimental results for TPC and TAA showed a great similarity.     

响应面法优化油菜秸秆真空热解液化工艺及生物油分析1

以油菜秸秆为原料,采用真空热解系统进行了制取生物油的中心组合实验研究,以热解终温、体系压力和升温速率为实验因子,生物油产率为实验指标,利用响应面法(RSM)对制备生物油的工艺参数进行了优化,并对在最优条件下制取的生物油进行了理化特性和化学组成分析。研究结果表明,热解终温、体系压力和升温速率对生物油产率有显著影响,热解终温和升温速率之间的交互作用显著;获得最佳热解液化工艺条件为:热解终温490.0℃、体系压力5.0 k Pa、升温速率20.0℃·min-1,在此条件下,生物油产率可达41.65%。与预测值42.00%较为接近。油菜秸秆真空热解所得生物油的含水量为33.85%,热值为18.65 MJ·kg-1,常温下的运动黏度为4.16 mm2·s-1,密度和p H值分别为1.14g·cm·3和2.32;生物油成分较为复杂,其中多种有机物可被进一步提取用作化工原料;生物油中羧酸、醛、酮类等腐蚀性和不稳定组分含量较高,需对其进一步精制,以提高其稳定性。     

Utilization of response surface methodology to optimize the culture media for the production of rhamnolipids by Pseudomonas aeruginosa AT10

Pseudomonas aeruginosa AT10 produced a mixture of surface‐active rhamnolipids when cultivated on mineral medium with waste free fatty acids as carbon source. The development of the production process to an industrial scale included the design of the culture medium. A 2⁴ full factorial, central composite rotational design and response surface modelling method (RSM) was used to enhance rhamnolipid production by Pseudomonas aeruginosa AT10. The components that are critical for the process medium were the carbon source, the nitrogen source (NaNO₃), the phosphate content (K₂ HPO₄/KH₂PO₄ 2:1) and the iron content (FeSO₄·7H₂O). Two responses were measured, biomass and rhamnolipid production. The maximum biomass obtained was 12.06 g dm^?3 DCW, when the medium contained 50 g dm^?3 carbon source, 9 g dm^?3 NaNO₃, 7 g dm^?3 phosphate and 13.7 mg dm^?3 FeSO₄·7H₂O. The maximum concentration of rhamnolipid, 18.7 g dm^?3, was attained in medium that contained 50 g dm^?3 carbon source, 4.6 g dm^?3 NaNO₃, 1 g dm^?3 phosphate and 7.4 mg dm^?3 FeSO₄·7H₂O. © 2002 Society of Chemical Industry