Optimization of Xylanase Production by Sporotrichum thermophile Using Corn Cobs and Response Surface Methodology

A 3² central composite experimental design was performed with the aim of optimizing the production of xylanase by Sporotrichum thermophile grown on corn cobs in submerged cultures. Various carbon and nitrogen sources were consecutively optimized, and corn cobs and ammonium phosphate concentrations were selected as substrates to test the effect of two variables, i.e., both substrate concentrations, on xylanase production. A second‐order quadratic model and a response surface method showed that the optimum conditions for xylanase production were 2.7 % [w/v] corn cobs and 0.7 % [w/v] ammonium phosphate. These optimum conditions were transferred to 7 L bioreactors, where activities as high as 56 U/mL were obtained.     

响应面法对小球藻Chlorella zofingiensis高产虾青素条件的优化

采用析因设计法(Plackett-burman)对影响Chlorella zofingiensis高产虾青素的相关因素进行评价,发现硝酸钠、光照强度、二价铁离子及醋酸钠浓度对虾青素产量影响显著.利用中心组合设计(central composite design)及响应面分析对影响虾青素产量的关键因素做进一步的优化,得到较佳的试验点为二价铁离子浓度0.41 mmol/L,硝酸钠浓度0.8 mmol/L,醋酸钠浓度37.1 mmol/L,光照强度650 E/m2×s.优化后虾青素产量从7.890mg/L提高到19.81mg/L,比优化前提高了2.5倍.     

响应面法优化酿酒酵母产油脂条件

运用响应面法对酿酒酵母(Saccharomyces cerevisiae)产油脂以及发酵条件优化进行了研究。首先根据单因素实验结果,利用Plackett-Burman设计对影响其产油脂相关因素进行评估并筛选出具有显著效应的3个因素:柠檬酸,CaCl2和初始pH值。接着用最陡爬坡试验逼近以上3个因子的最大响应区域后,采用Box-Behnken设计以及响应面分析法,确定其优化后发酵条件为(w/v):葡萄糖15%,蛋白胨0.2%,酵母浸粉0.4%,柠檬酸0.471%,MgSO4·7H2O0.1%,ZnSO4·7H2O0.2%,CaCl20.025%,FeSO4·7H2O0.005%,初始pH值为6.74,180r/min,30°C培养96h。优化后的油脂产率(干重)达到14.55%,比在种子培养基中油脂产率4.76%提高了2倍左右。     

Enhanced chrysene degradation by halotolerant Achromobacter xylosoxidans using Response Surface Methodology

Degradation of chrysene, a four ring High Molecular Weight (HMW) Polycyclic Aromatic Hydrocarbon (PAH) is of intense environmental interest, being carcinogenic, teratogenic and mutagenic. Multiple PAH degrading halotolerant Achromobacter xylosoxidans was isolated from crude oil polluted saline site. Response Surface Methodology (RSM) using Central Composite Design (CCD) of Bushnell-Haas medium components was successfully employed for optimization resulting 40.79% chrysene degradation on 4th day. The interactions between variables as chrysene and glucose concentrations, pH and inoculum size on degradation were examined by RSM. Under optimum conditions, A. xylosoxidans exhibited 85.96% chrysene degradation on 5th day. The optimum values predicted by RSM were confirmed through confirmatory experiments. It was also noted that pH and glucose as co-substrate play a dynamic role in enhancement of chrysene degradation. Hence, A. xylosoxidans can be further used for subsequent microcosm and in situ experiments for its potential to remediate PAH contaminated saline and non-saline soils.     

响应面法优化枯草芽孢杆菌产脂肪酶的合成培养基

对枯草芽孢杆菌(Bacillus subtilis)CICC20034利用合成培养基液体发酵产脂肪酶的条件进行了优化。首先采用单因子实验筛选出最适诱导剂为三丁酸甘油酯,氮源为尿素,碳源为葡萄糖,无机盐为MgSO4。在此基础上,利用Plackett-Burman设计对影响产酶因素的效应进行评价,筛选出具有显著效应的三丁酸甘油酯、尿素、KH2PO4和培养基起始pH值4个最显著的因素。用最陡爬坡路径逼近最大产酶区域后,利用响应面中心组合设计对显著因素进行优化,获得最适合成培养基组分为:葡萄糖8g/L,尿素8.57g/L,三丁酸甘油酯2.62%,KH2PO42.59g/L,MgSO4.7H2O0.5g/L,TritonX-1000.5g/L,pH9.47。优化后的B.subtilis CICC 20034胞外脂肪酶活力达0.483U/ml,比初始酶活力0.072U/ml提高了6.7倍。     

丛梗孢酵母发酵产赤藓糖醇的响应面优化

为了提高丛梗孢酵母发酵产赤藓糖醇的产量,在前期单因素实验结果的基础上,利用Plackett-Burman实验设计对影响其产赤藓糖醇的发酵条件进行评估并筛选出了影响显著的3个因素:葡萄糖、初始pH和温度.采用响应面法进行实验方案设计,利用SAS软件对其结果进行二次回归分析,确定了优化后的发酵条件为:葡萄糖260g/L、酵...     

Ectoine Production by Halomonas boliviensis: Optimization Using Response Surface Methodology

Two cultivation steps were used for production of biomass and ectoine by Halomonas boliviensis, respectively. The optimization of some nutrient parameters in each step was investigated by using response surface methodology. Twenty and 12 experiments were performed to attain optimal conditions for biomass and ectoine production, respectively. The model predicted a maximum biomass concentration of 3.34 g/L on optimization of NH₄Cl, K₂HPO₄, and MgSO₄•7H₂O concentrations during the first cultivation, while a maximum ectoine concentration of 1.27 g/L was predicted on optimizing NaCl and monosodium glutamate concentrations in the second cultivation. The experimental values obtained (3.36 g biomass/L and 1.25 g ectoine/L) were in good agreement with the predicted values. The optimized conditions were also used for two-step 1.5-L fed-batch fermentations. In the first step, biomass concentration of 28.7 g/L was obtained while in the second step biomass concentration increased to 63 g/L. Ectoine concentration of 9.2 g/L was obtained, and the overall ectoine productivity was 6.3 g/L/day, being among the highest reported so far.     

Optimization of Chlorpyrifos Degradation by Assembled Bacterial Consortium Using Response Surface Methodology

Chlorpyrifos is a commonly used organophosphate pesticide. Its extensive use and associated serious soil and water contamination have gained increasing environmental concern. Biodegradation is a promising way to remediate chlorpyrifos contamination. There are many reports on various chlorpyrifos degrading microorganisms, but only a few on biodegradation of chlorpyrifos by consortia. Hence, the present study attempted to assemble a novel bacterial consortium C5 for the biodegradation of chlorpyrifos. The 16S rRNA gene-based molecular analysis revealed that the bacterial consortium consisted of Staphylococcus warneri CPI 2, Pseudomonas putida CPI 9 and Stenotrophomonas maltophilia CPI 15. Optimization of chlorpyrifos degradation by the consortium C5, using a Box–Behnken design, was carried out taking into account four important variables: temperature, pH, the initial concentration of chlorpyrifos and time of incubation. C5 is capable of giving 90% degradation of chlorpyrifos (125 ppm) in 8 days of incubation under optimized conditions of pH (7) and temperature (30°C). Growth curve and degradation study under optimized conditions confirmed that consortium could improve the biodegradation potential. From these results, we conclude that the novel consortium C5 of three species can be used to eliminate chlorpyrifos from various environmental compartments and can be implemented in bioreactors in a cost-effective, safe and environmentally friendly manner.     

Enhanced Sono-Fenton-Like Oxidation of PAH-Contaminated Soil Using Nano-Sized Magnetite as Catalyst: Optimization with Response Surface Methodology

In the current study, Fe₃O₄ NPs were synthesized and used as catalysts in a sono-Fenton-like process for remediation of polycyclic aromatic hydrocarbon (PAH)-contaminated soil. The effects of operational variables were studied using central composite design (CCD) optimization approach. Results indicated that the effects of H₂O₂ concentration, Fe₃O₄ NPs dosage, ultrasonic power and pH were significant for pyrene removal as a contaminant model. In optimum experimental conditions, including H₂O₂ concentration of 78 mM, Fe₃O₄ NPs dosage of 18 mM, ultrasonic power of 313 W and pH value of 3.46, the observed pyrene removal was obtained 98.37%, which was verified through the additional experimental tests (99.33%). Pseudo first-order kinetic model was well fitted with the experimental data of pyrene removal with significant coefficient of correlation (R²: 0.9672). Accordingly, an unwashed real soil sample containing diffident PAHs (pyrene, flurene, acenaphthylene, phenenthrene, chrysene, etc) was subjected to sono-Fenton-like process based on optimized conditions. The obtained findings revealed that the removal (%) ranged between 37.7% and 85.19% for different PAHs.     

Enhancement of Copper Dissolution from a Sulfide Ore by Using Thiobacillus thiooxidans

Copper dissolution from a sulfide ore (with covellite as the main copper phase) was investigated in cultures of Thiobacillus ferrooxidans or Thiobacillus thiooxidans and in abiotic controls. In unsupplemented media, T. ferrooxidans was more efficient than T. thiooxidans. In the presence of ferric iron, the dissolution of covellite was not significantly different in cultures inoculated with T. ferrooxidans or T. thiooxidans. However, the most extraction was found in T. thiooxidans cultures supplemented with ferrous sulfate. The first results were explained by the mechanism proposed by Schippers and Sand (Appl Envir Microbiol 65:319-321, 1999), which involves polysulfides and sulfur as intermediates. This mechanism was extended to explain the behavior of T. thiooxidans culture supplemented with ferrous iron.     

响应面分析法优化超声提取蓖麻碱工艺

以蓖麻叶为原料,对蓖麻碱的超声提取工艺优化进行研究,在单因素试验的基础上,选择超声时间、超声功率、料液比为自变量,以蓖麻碱提取率为影响值,采用响应面试验设计方法,研究各自变量及其交互作用对蓖麻碱提取率的影响。利用Design Expert8软件得到回归方程得模型并进行响应面分析,确定超声提取蓖麻碱的最佳工艺条件为料水比为1∶25 g/mL,超声时间为103.03 min,超声功率为621.05 W,此条件下蓖麻碱的提取率为2.63‰。     

Optimization of Phosphatidylinositol-specific Phospholipase C Production Using Response Surface Methodology

Summary Response surface methodology was employed in optimizing the nutrient levels needed towards the optimal production of phosphatidylinositol-specific phospholipase C enzyme by Bacillus thuringiensis serovar. kurstaki. A 2³ factorial central composite experimental design was used. The multiple regression equation, relating the enzyme activity to the nutrient medium, was used to find the optimum values of glucose, peptone and dipotassium hydrogen phosphate. The optimum values of these variables for maximal enzyme production were found to be: glucose, 6.5 g l⁻¹; peptone, 5.38 g l⁻¹ and dipotassium hydrogen phosphate, 6.36 g l⁻¹ with the predicted enzyme activity of 0.96 U ml⁻¹.     

Optimization of 1-Deoxynojirimycin Extraction from Mulberry Leaves by Using Response Surface Methodology

Mulberry 1-deoxynojirimycin (DNJ, a potent α-glycosidase inhibitor) has therapeutic potency against diabetes mellitus. However, the amount of DNJ in mulberry leaves is low (about 0.1%), and a more effective extraction method is needed. Ultrasound-assisted extraction (UAE) was applied in this study for mulberry DNJ extraction, and five factors, the percentage of ethanol in the extraction solvent (x ₁), ratio of the extraction solvent to mulberry sample (x ₂), ultrasonic power (x ₃), extraction temperature (x ₄) and extraction time (x ₅), were investigated by fractional factorial 2^(5-1) design (FFD) to obtain the optimum extraction efficiency (DNJ yield, Y ₁) and extraction productivity (total yield, Y ₂). The results showed that x ₂, x ₃ and x ₅ had significant impact on Y ₁ and Y ₂, and were further optimized by response surface methodology (RSM). Under the optimized conditions (x ₂, x ₃ and x ₅ of 7 ml/g, 180 W and 260 s, respectively), DNJ-enriched powder (0.8%) was produced with high extraction efficiency (98%) and productivity (20%), enabling this product to be used for nutraceutical purposes.     

利用响应面法优化红谷霉素发酵培养基*

在摇瓶条件下,对链霉菌702发酵生产过程中的主要培养基组成对产红谷霉素影响进行的研究。试验采用响应面法优化摇瓶发酵培养基,利用全因子实验设计筛选出对链霉菌702产红谷霉素重要影响因子黄豆饼粉和工业蛋白胨,应用最陡爬坡实验法接近重要因子的最优水平,然后应用中心复合设计确定重要因子的最优水平。优化后的培养基组成为：玉米淀粉20g,玉米粉20g,葡萄糖20g,磷酸二氢钾0．3g,蛋白胨9g,黄豆饼粉23g,硝酸钾2．5g,硫酸铵2．5g,豆油5mL,氯化钠3g,碳酸钙6g,定容至1L。实验结果表明,采用优化后的培养基,其发酵液红谷霉素效价达到1,500μg／mL,比优化前提高了3.08倍。     

响应面法对Acidithiobacillus ferrooxidans发酵液脱除H2S的优化研究

目的:利用Acidithiobacillus ferrooxidans(A.f)发酵液对H2S的脱除条件进行优化.方法:在前期单因素影响脱硫率的基础上,利用Design expert软件中Box-Benhnker中心组合实验设计和响应面分析法对A.f发酵液脱除H2S的气体流量、温度和进行进一步优化.结果:获得了优化的A.f发酵液脱除H2S工艺,反应温度为31.02℃,气体流量为40.03 ml/min,H2S浓度为8.17g/m3.结论:在该条件下脱硫率达99.37％,表明响应面法可有效用于A.f发酵液脱除H2S工艺的优化.     

响应面分析法优化(R)-扁桃酸发酵培养基

采用响应面分析法对Bacillussp.HB20菌株合成(R)-扁桃酸的培养基成分进行优化。首先利用Plackett-Burman试验设计筛选出影响(R)-扁桃酸产率的三个主要因素:麦芽糖、蛋白胨和牛肉膏。在此基础上用最陡爬坡路径逼近最大响应区域,再利用Box-Behnken试验设计及响应面分析法进行回归分析。结果表明,麦芽糖、蛋白胨和牛肉膏浓度与(R)-扁桃酸产率存在显著的相关性,通过求解回归方程得到最佳质量浓度:蛋白胨11.507g/L,牛肉膏6.708g/L,麦芽糖10.907g/L,(R)-扁桃酸产率理论最大值达到66.87%。经模型验证,预测值与验证试验平均值接近,在优化条件下(R)-扁桃酸产率提高了25.87%。     

Optimization of PEGylation Conditions for BSA Nanoparticles Using Response Surface Methodology

Chemical coupling of polyethylene glycol (PEG) to proteins or particles (PEGylation), prolongs their circulation half-life by greater than 50-fold, reduces their immunogenicity, and also promotes their accumulation in tumors due to enhanced permeability and retention effect. Herein, phase separation method was used to prepare bovine serum albumin (BSA) nanoparticles. PEGylation of BSA nanoparticles was performed by SPA activated mPEG through their free amino groups. Effect of process variables on PEGylation efficiency of BSA nanoparticles was investigated and optimized through response surface methodology with the amount of free amino groups as response. Optimum conditions was found to be 32.5 g/l of PEG concentration, PEG-nanoparticle incubation time of 10 min, incubation temperature of 27°C, and pH of 7 for 5 mg of BSA nanoparticles in 1 mL phosphate buffer. Analysis of data showed that PEG concentration had the most noticeable effect on the amount of PEGylated amino groups, but pH had the least. Mean diameter and zeta potential of PEGylated nanoparticles under these conditions were 217 nm and −14 mV, respectively. In conclusion, PEGylated nanoparticles demonstrated reduction of the negative surface charge compared to the non modified particles with the zeta potential of −31.7 mV. Drug release from PEGylated nanoparticles was almost slower than non-PEGylated ones, probably due to existence of a PEG layer around PEGylated particles which makes an extra resistance in opposition to drug diffusion.     

利用响应面法优化Bacillus natto TK-1产脂肽发酵培养基

用响应面法对Bacillus natto TK-1发酵产脂肽的培养基进行了优化。首先用Plackett-Burman法筛选出三个影响较大的重要因素,分别为：蛋白胨、酵母粉、CaCl2 ,然后进行最陡爬坡实验逼近最佳响应面区域,最后通过Box-Behnken设计,利用SAS软件进行回归分析,得到各因素的最佳浓度。在优化培养基条件下,发酵液的溶血圈直径较初始培养基提高了29.3 %,脂肽的产量提高了约30%。     

A glycerol-free process to produce biodiesel by supercritical methyl acetate technology: An optimization study via Response Surface Methodology

In this study, fatty acid methyl esters (FAME) have been successfully produced from transesterification reaction between triglycerides and methyl acetate, instead of alcohol. In this non-catalytic supercritical methyl acetate (SCMA) technology, triacetin which is a valuable biodiesel additive is produced as side product rather than glycerol, which has lower commercial value. Besides, the properties of the biodiesel (FAME and triacetin) were found to be superior compared to those produced from conventional catalytic reactions (FAME only). In this study, the effects of various important parameters on the yield of biodiesel were optimized by utilizing Response Surface Methodology (RSM) analysis. The mathematical model developed was found to be adequate and statistically accurate to predict the optimum yield of biodiesel. The optimum conditions were found to be 399 °C for reaction temperature, 30 mol/mol of methyl acetate to oil molar ratio and reaction time of 59 min to achieve 97.6% biodiesel yield.     

Optimization of Culture Conditions for Fermentation of Soymilk Using Lactobacillus casei by Response Surface Methodology

Soymilk was fermented with Lactobacillus casei, and statistical experimental design was used to investigate factors affecting viable cells of L. casei, including temperature, glucose, niacin, riboflavin, pyridoxine, folic acid and pantothenic acid. Initial screening by Plackett-Burman design revealed that among these factors, temperature, glucose and niacin have significant effects on the growth of L. casei. Further optimization with Box-Behnken design and response surface analysis showed that a second-order polynomial model fits the experimental data appropriately. The optimum conditions for temperature, glucose and niacin were found to be 15.77 °C, 5.23 and 0.63 g/L, respectively. The concentration of viable L. casei cells under these conditions was 8.23 log₁₀ (CFU/mL). The perfect agreement between the observed values and the values predicted by the equation confirms the statistical significance of the model and the model’s adequate precision in predicting optimum conditions.