响应面法优化重组工程菌的发酵工艺条件

目的：对基因改造运动发酵单胞菌的发酵工艺条件进行优化,提高重组菌发酵乙醇产量。方法：使用分子克隆实验操作技术构建重组运动发酵单胞菌,以单因素实验为基础,利用Box-Behnken中心组合实验和响应面分析法,确定了影响重组菌高产乙醇的三个重要因素。结果：成功构建含有YfdZ、MetB基因和Hsp基因的重组菌Zymomonas mobilis HYM,发酵主要影响因素的最佳条件分别为温度28℃,葡萄糖浓度24%（W/V）,pH7.4。在此优化条件下,Zymomonas mobilis HYM的乙醇产量可高达105.0735g/l,比原始菌株乙醇产量提高16.4%。结论：用中心组合设计和响应面分析法优化重组运动发酵单胞菌的发酵工艺条件,显著提高乙醇产量。     

响应面法优化脯氨酸羟化酶转化反应工艺条件

通过优化脯氨酸羟化酶表达条件和转化反应条件,提高其转化反应效率。采用单因素法筛选脯氨酸羟化酶的最佳诱导温度、诱导剂浓度和转化反应条件,并采用响应面法预测影响转化反应各因素的最佳条件。结果显示,经过筛选和验证,蛋白表达最适诱导温度为28℃,IPTG浓度为0.2 mmol/L;转化反应最佳条件为:120 mmol/L 2-(N-吗啡啉)乙磺酸(pH 6.6)、1.5% Nonidet P-40、200 mmol/L L-脯氨酸、200 mmol/L α-酮戊二酸,6 mmol/L L-抗坏血酸、6.0 mmol/L 硫酸亚铁,最适反应温度为27℃,振荡速率为152 r/min。在最佳条件下,转化反应进行48 h后产物反式-4-羟基-L-脯氨酸的转化率可达100%,为合成反式-4-羟基-L-脯氨酸奠定了坚实的实验基础。     

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.     

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。     

利用响应面法优化石榴酒发酵工艺条件

攀枝花的石榴品种繁多，籽粒味甜汁多，为了充分利用这一优势资源，本文进行石榴酒发酵，并对其发酵工艺利用响应面分析法进行优化。以酒精度为指标，考虑发酵温度、二氧化硫添加量、酵母接种量对石榴酒发酵酒精度的影响，然后根据中心组合(Box-Benhnken)原理采用三因素三水平的分析法，依据回归确定各工艺条件的影响因素，以石榴酒发酵酒精度为响应面和等高线，分析各个因素的显著性和交互作用，结果表明优化得到石榴酒发酵的最佳工艺条件为二氧化硫添加量51.3 mg/kg，酵母菌接种量5.33%，发酵温度24.98℃，酒酒精度的理论值为9.58%。通过响应面分析优化发酵工艺，为今后石榴酒发酵的更进一步开发提供一定的理论依据。     

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.     

利用响应面法优化L-组氨酸摇瓶发酵培养基

通过单因素实验对L-组氨酸产生菌LGS4的发酵培养基成分进行筛选,确定了3个影响较大的重要因素,即酵母膏、尿素、硫酸镁。在此基础上,采用二次响应面分析法进行回归分析,得到各因素的最佳水平值。经5批培养验证,预测值与验证试验平均值接近。优化后培养基组成为(g.L-1):蔗糖150,硫酸铵50,酵母膏10,尿素1.2,MgSO42.2,KH2PO41.5,K2HPO40.5,Na2HPO40.5。优化后的发酵培养基使LGS4菌株的L-组氨酸产量提高了15.25%。     

响应面法优化灵芝菌丝体胞内灵芝酸的提取

利用单因子试验和响应面法优化了影响灵芝菌丝体灵芝酸的提取过程。基于3因素3水平的中心组合设计,得到了描述胞内灵芝酸提取得率与操作参数之间的二次响应面模型。在乙醇浓度为93.6%(v/v)、提取温度79.1℃、提取液固比42.2∶1、提取次数2次、每次提取时间2 h的条件下,最大灵芝酸理论提取得率为28.36 mg胞内灵芝酸每克干菌丝。模型优化条件下的实际灵芝酸提取得率(28.72 mg/g干菌丝)与理论灵芝酸提取得率(28.36 mg/g干菌丝)相符。     

Optimization Studies on Gastroretentive Floating System Using Response Surface Methodology

The aim of the present investigation was to develop and optimize gastroretentive floating system of amoxicillin for the efficient treatment of peptic ulcer induced by Helicobacter pylori infection. Floating microballoons were developed using central composite design (CCD), and optimization was done by employing response surface methodology. The selected independent variables were cellulose acetate phthalate, drug–Eudragit S100 ratio, and the ratio of dichloromethane/ethanol/isopropyl alcohol. The selected dependent variables were yield, mean particle size, buoyancy, encapsulation efficiency, and drug release within 8 h. A quadratic polynomial model was generated which had linear, interaction, and quadratic terms to predict and evaluate the independent variables with respect to the dependent variables. Results showed that selected independent variables significantly affect the yield (30.53–82.71%), particle size (31.62–47.03 μm), buoyancy (42.68–95.75%), encapsulation efficiency (56.96–93.13%), and cumulative drug release from the microballoons (34.01–74.65%). The interaction and quadratic terms were also found to affect the process variables. An excellent agreement was found between the actual value and predicted value. In conclusion, it can be said that CCD is a valuable second-degree design to develop and optimize GFS of amoxicillin which in turn provides a basis to localize the drug release in the gastric region for effective treatment of H. pylori-mediated infection.     

响应面法优化荞麦胰蛋白酶抑制剂固定化条件

目的:以大肠杆菌表达的重组荞麦胰蛋白酶抑制剂为原材料,研究其固定化方法及条件。方法:以0.2%聚乙烯醇-3%海藻酸钠溶液为载体,CaCl2为固定剂,用物理包埋法对荞麦胰蛋白酶抑制剂进行固定化;在CaCl2浓度、载体与抑制剂体积比以及固定化时间3个单因素基础上,利用响应面法对荞麦胰蛋白酶抑制剂固定化的影响因素进行优化。结果:建立了响应面法优化固定荞麦胰蛋白酶抑制剂的模型,经优化后得到如下最佳固定化条件:CaCl2浓度为5.5%,载体与抑制剂体积比为1.6∶1,固定化时间为31min。在此条件下实际测得固定化抑制剂抑制率为72.4%,而模型预测此条件下的抑制率为74.3%,实测值与理论值相差很小。结论:所建模型拟合程度较高,用该模型优化荞麦胰蛋白酶抑制剂固定化的工艺条件参数准确可信,可为进一步开发胰蛋白酶的应用提供重要参考。     

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。     

响应面法优化紫甘薯花色苷水解条件

使用响应面法优化了紫甘薯花色苷的水解条件.使用高效液相色谱-多波长检测器测定花青素含量.采用3因素3水平Box-Behnken中心组合设计,优化了水解时间、盐酸浓度和水解温度对矢车菊素和芍药素峰面积之和的综合影响.结果显示,紫甘薯花色苷的最佳水浴水解条件为:盐酸浓度4.54 mol/L,水浴温度100℃,水解时间45.8 min.在最优条件下进行验证试验,响应值为理论值的96.5％,说明该响应面法优化方法可行.     

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.     

响应面优化-超声提取黄姜中总黄酮工艺研究

目的:利用响应面法对黄姜总黄酮的提取工艺进行优化.方法:在单因素的基础上以超声功率、超声时间、料液比、乙醇浓度为响应因素,黄姜总黄酮的提取率为响应值,根据中心组合实验原理,采用四因素三水平的响应面分析法.结果:确定提取最佳工艺参数为超声时间29.53 min、超声功率为400.76w、料液比为1:42.19、乙醇浓度为79.60％,在此条件下总黄酮提取率的预测值为11.15 (mg/g).结论:考虑到实际操作的问题,将提取最佳工艺参数修正为超声时间30 min、超声功率400w、料液比1:40、乙醇浓度80％,进行3次平行试验,结果总黄酮提取率为11.09 mg/g,绝对误差为0.057％,说明实验结果与模型吻合很好.     

响应面分析法优化当归粗多糖提取工艺

选取岷县当归药材为原料,采用水提醇沉法进行多糖提取,以当归粗多糖得率为指标,探讨加水量、回流提取时间、水提液的浓缩比、醇沉后所达含醇比例对当归多糖得率的影响。在单因素分析的基础上,采用响应面分析法(RSM)确定了当归粗多糖最佳提取条件为:加水量837.6 mL/100 g,浓缩后溶液体积为228.12 mL,最终含乙醇浓度为65.80%,回流提取时间2 h,在此条件下预测当归粗多糖得率理论的最佳值为10.44%,实际验证值为10.40%,两者相符,说明RSM法分析的可靠性。     

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.     

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.