响应面分析法优化重组大肠杆菌生物合成谷胱甘肽的条件

通过响应面分析法和典型性分析得出重组大肠杆菌酶法合成谷胱甘肽的最优条件:菌体量249 mg/mL,磷酸钾缓冲液145 mmol/L,MgCl243 mmol/L和ATP 34 mmol/L,预测谷胱甘肽最大量为16.50 mmol/L。验证性实验证明在优化条件下,重组大肠杆菌酶法合成谷胱甘肽达16.42 mmol/L。响应面分析还表明,在重组大肠杆菌酶法合成谷胱甘肽各因素中,MgCl2和ATP,以及菌体量与磷酸钾缓冲液之间的交互作用较显著。     

Optimization of low-cost biosurfactant production from agricultural residues through response surface methodology

Biosurfactants are surface-active compounds capable of reducing surface tension and interfacial tension. Biosurfactants are produced by various microorganisms. They are promising replacements for chemical surfactants because of biodegradability, nontoxicity, and their ability to be produced from renewable sources. However, a major obstacle in producing biosurfactants at the industrial level is the lack of cost-effectiveness. In the present study, by using corn steep liquor (CSL) as a low-cost agricultural waste, not only is the production cost reduced but a higher production yield is also achieved. Moreover, a response surface methodology (RSM) approach through the Box–Behnken method was applied to optimize the biosurfactant production level. The results found that biosurfactant production was improved around 2.3 times at optimum condition when the CSL was at a concentration of 1.88 mL/L and yeast extract was reduced to 25 times less than what was used in a basic soybean oil medium (SOM). The predicted and experimental values of responses were in reasonable agreement with each other (Pred-R² = 0.86 and adj-R² = 0.94). Optimization led to a drop in raw material price per unit of biosurfactant from $47 to $12/kg. Moreover, the biosurfactant product at a concentration of 84 mg/L could lower the surface tension of twice-distilled water from 72 mN/m to less than 28 mN/m and emulsify an equal volume of kerosene by an emulsification index of (E24) 68% in a two-phase mixture. These capabilities made these biosurfactants applicable in microbial enhanced oil recovery (MEOR), hydrocarbon remediation, and all other petroleum industry surfactant applications.     

Optimization of laccase production from a novel strain-Streptomyces psammoticus using response surface methodology

Response surface methodology was employed for the optimization of different nutritional and physical parameters for the production of laccase by the filamentous bacteria Streptomyces psammoticus MTCC 7334 in submerged fermentation. Initial screening of production parameters was performed using a Plackett - Burman design and the variables with statistically significant effects on laccase production were identified. Incubation temperature, incubation period, agitation rate, concentrations of yeast extract, MgSO(4)7H(2)O, and trace elements were found to influence laccase production significantly. These variables were selected for further optimization studies using a Box-Behnken design. The statistical optimization by response surface methodology resulted in a three-fold increase in the production of laccase by S. psammoticus MTCC 7334.     

Optimization and evaluation of acetylcholine esterase immobilization on ceramic packing using response surface methodology

In the present attempt a method for the immobilization of acetylcholine esterase (AChE) was developed. In this method, the enzyme was immobilized onto a ceramic cylinder support using a sol–gel–multiwall carbon nanotube (MWCNT) composite. Response surface methodology (RSM) was used for the design and analysis of immobilization experiments. Quadratic mathematical model equations were derived for the prediction of enzyme activity. Then the effects on enzyme activity at 30, 40 and 50 min after process initiation of varying each of two parameters over five levels were investigated. These parameters were the AChE:MWCNT ratio (X₁), and AChE–MWCNT:sol–gel ratio (X₂). The optimum values of X₁ and X₂ for the immobilization of AChE on ceramic packing were found to be 1.07 and 0.43, respectively. Using these optimum parameters it was shown that enzyme immobilization with MWCNTs and sol–gel was more effective than immobilization with sol–gel or graphite and sol–gel. Scanning electron microscopic (SEM) images revealed a porous surface comprised of MWCNT–AChE encapsulated in sol–gel. Furthermore, the system was highly reproducible with standard deviations after three successive assays of 1.88%, 2.11% and 2.13% at 30, 40 and 50 min after process initiation, respectively.     

Optimization of fermentation parameters in phage production using response surface methodology

Previously, we used computer-controlled fermentation technology to improve the yield of filamentous phage produced in Escherichia coli by 10-fold (Grieco et?al., Bioprocess Biosyst Eng 32:773-779, 2009). In the current study, three major fermentation parameters (temperature, dissolved oxygen [DO], and pH) were investigated using design of experiments (DOE) methodology. Response surface methodology (RSM) was employed to create a process model and determine the optimal conditions for maximal phage production. The experimental data fitted best to a quadratic model (p?相似文献   

响应面法对红法夫酵母合成虾青素主要影响因素的优化

在单因素试验确定了红法夫酵母生物合成虾青素培养基组份的基础上,用响应面法对其浓度进行优化。首先用分式析因设计评价了培养基的各组份对虾青素产量的影响,并找出主要影响因子为蔗糖和酵母粉,二者分别达到了极显著和显著水平。用最陡爬坡路径逼近最大响应区域后,运用旋转中心复合设计及响应面分析,确定了主要影响因子的最佳浓度。其中,蔗糖的最佳浓度为49.8g/L,酵母粉的浓度为9.6g/L。菌株在优化培养基中的虾青素产量为9861μg/L,比优化前增加了近1倍。     

产S-酰胺酶培养基统计学筛选与响应面优化

利用Design Expert软件中的两水平实验设计和响应面法,对发酵生产S-酰胺酶(可用于拆分2,2-二甲基环丙甲酰胺外消旋体)的培养基进行了优化。采用Plackett-Burman(PB)设计对培养基中相关影响因素的效应进行评价并筛选出了有显著效应的葡萄糖、酵母粉及2,2-二甲基环丙甲酰胺浓度,其他因素对酰胺酶产量的影响不显著。然后用旋转中心组和实验设计及响应面分析确定了主要影响因素的最佳条件,在优化的培养基中,酰胺酶产量达到168 U/L,比优化前的80 U/L提高了110.0%。     

响应面-满意度函数优化重组大肠杆菌产NMN转移酶发酵条件

重组大肠杆菌Escherichaia coli能高效表达NMN转移酶,以此为出发菌株,以菌体生长量OD600和NMN转移酶的活力为响应值,对重组大肠杆菌产NMN转移酶的发酵条件进行优化.首先以Plackett-Burman实验设计优化筛选出3个主要影响因子:胰蛋白胨、甘油、MgSO4;随后以Box-Behnken中心组合设计建立上述3个因子对OD600和NMN转移酶活力水平的数学模型;最后通过满意度函数获得最佳发酵条件为:酵母粉30 g/L,胰蛋白胨10.5 g/L,甘油3.49 mL/L,MgSO40.45 g/L,K2 HPO440.5 g/L,KH2 PO46.0 g/L,NH4 Cl 1.5 g/L,NaCl 0.6 g/L,接种量1.5%,诱导时间12 h.在该优化条件下,菌体生长和产酶水平均获得了显著的提升.重组NMN转移酶的活力水平从8.85 U/mg提高到15.48 U/mg,菌体生长量OD600从4.85提高到6.01,提高幅度分别为74.92%和23.92%.     

Optimization of microbiological parameters for enhanced griseofulvin production using response surface methodology

Central composite design was used to determine the optimal levels of microbiological parameters, viz., slant age, seed age and inoculum level, for enhanced griseofulvin production by Penicillium griseofulvum MTCC 1898 and Penicillium griseofulvum MTCC 2004 in shake flask fermentation. The optimal levels of slant age, seed age and inoculum level for Penicillium griseofulvum MTCC 1898 were found to be 8.8772 days, 4.2093 days, 12% (v/v) (᷁.56 kg dry cell mass/m³) and for Penicillium griseofulvum MTCC 2004, 8.221 days, 3.4875 days and 9% (v/v) (̀.09 kg dry cell mass/m³) respectively. The yield of griseofulvin under optimal conditions was found to be 1.65 times for Penicillium griseofulvum MTCC 1898 and 1.07 times for Penicillium griseofulvum MTCC 2004 higher than that obtained using unoptimized conditions. The fermentation time for maximum production of griseofulvin by Penicillium griseofulvum MTCC 1898 and Penicillium griseofulvum MTCC 2004 decreased by 4 days and 2 days respectively.     

Optimization of growth medium for the production of spores from Bacillus thuringiensis using response surface methodology

The effects of cultivation medium compositions including tapioca, fishmeal, CaCO₃ and (NH₄)₂SO₄ for the growth of Bacillus thuringiensis YMB 96-1988 were accessed by using response surface methodology (RSM). The two-level (2^4–1) fractional factorial designs (FFD) which involve two concentrations of each nutrient, and the paths of steepest ascent were effective in searching for the major factors of the bacteria growth. This allows the fitting of a first order linear model to the data. In this study, supplementary CaCO₃ showed a negative effect on the spore production based on the first order regression coefficients derived from SAS programme. Subsequently, a 2³ central composite design (CCD) was used for allocation of treatment combinations. Preliminary studies showed that tapioca and fishmeal is believed to be the major factors for the growth of B. thuringiensis. Estimated optimum compositions for the production of spores by B. thuringiensis are as follows: tapioca, 5.01%; fishmeal 5.86%; (NH₄)₂SO₄ 0.06% and resulted in a maximum spore count of 8.56?×?10⁸/ml was obtained. This value is close to the 8.35?×?10⁸/ml spore density as counted from actual experimental observations.     

Optimization of fermentation conditions for the production of ethanol from sago starch using response surface methodology

The quantitative effects of temperature, pH and time of fermentation were investigated on simultaneous saccharification and fermentation (SSF) of ethanol from sago starch with glucoamylase (AMG) and Zymomonas mobilis ZM4 using a Box–Wilson central composite design protocol. The SSF process was studied using free enzyme and free cells and it was found that with sago starch, maximum ethanol concentration of 70.68 g/l was obtained using a starch concentration of 140 g/l, which represents an ethanol yield of 97.08%. The optimum conditions for the above yield were found to be a temperature of 36.74 °C, pH of 5.02 and time of fermentation of 17 h. Thus by using the central composite design, it is possible to determine the accurate values of the fermentation parameters where maximum production of ethanol occurs.     

Optimization of galacto-oligosaccharide production by Bifidobacterium infantis RW-8120 using response surface methodology

Oligosaccharide (OS) production, cell concentration (2×10⁹ colony-forming unit/ml), lactose concentration (25% wt/vol), reaction time (6 h), and temperature (50°C) were chosen as the central condition of the central composite design (CCD) for optimizing the production process using Bifidobacterium infantis RW-8120 in skim milk. Statistical analysis (P<0.01) revealed that the most relevant variable concerning OS production and yield was the lactose concentration. The coefficient of determination (R ²) is good for the second-order OS production model (0.92) and fairly good for the second-order nonlinear OS yield model (0.816). An increase of lactose concentration and temperature resulted in a higher OS production. The optimal values for OS production appear to be near the area associated with the central points of the modeling design except for the lactose concentration, which was 40% (wt/vol) of the final volume. Received 29 March 2002/ Accepted in revised form 09 August 2002     

Optimization of mycophenolic acid production in solid state fermentation using response surface methodology

Mycophenolic acid (MPA) can be produced in solid state fermentation. An isolate of Penicillium brevi-compactum ATCC 16024 grown on moist wheat bran produced a titre of 425 mg per kg of wheat bran. Central composite rotatable design and response surface methodology were employed to derive a statistical model for media optimization towards production of mycophenolic acid. Five levels with a five factorial design were adopted. The correlation coefficient was 0.82, ensuring a satisfactory adjustment of the model to the experimental values. This statistical design was very effective in improving the titre of mycophenolic acid up to 3286 mg per kg of wheat bran. Received 24 July 1998/ Accepted in revised form 4 December 1998     

响应面法优化Zn-Mn-Co/HZSM-5催化乙醇脱水制乙烯

采用响应面法研究温度、乙醇浓度、质量空速对锌、锰、钴改性的HZSM-5催化乙醇脱水制备乙烯过程中乙烯收率的影响。结果表明反应温度对乙烯收率影响最大,并且各因素之间存在交互作用。用响应面方法确定乙醇脱水制备乙烯的最佳工艺条件是：温度261.3 ℃,乙醇浓度34.4％,质量空速1.18 h?1,在该条件下乙烯收率达到98.69％。     

Optimization of culture medium for novel cell-associated tannase production from Bacillus massiliensis using response surface methodology

Naturally immobilized tannase (tannin acyl hydrolase, E.C. 3.1.1.20) has many advantages, as it avoids the expensive and laborious operation of isolation, purification, and immobilization, plus it is highly stable in adverse pH and temperature. However, in the case of cell-associated enzymes, since the enzyme is associated with the biomass, separation of the pure biomass is necessary. However, tannic acid, a known inducer of tannase, forms insoluble complexes with media proteins, making it difficult to separate pure biomass. Therefore, this study optimizes the production of cell-associated tannase using a "protein-tannin complex" free media. An exploratory study was first conducted in shake-flasks to select the inducer, carbon source, and nitrogen sources. As a result it was found that gallic acid induces tannase synthesis, a tryptose broth gives higher biomass, and lactose supplementation is beneficial. The medium was then optimized using response surface methodology based on the full factorial central composite design in a 3 l bioreactor. A 2(3) factorial design augmented by 7 axial points (alpha = 1.682) and 2 replicates at the center point was implemented in 17 experiments. A mathematical model was also developed to show the effect of each medium component and their interactions on the production of cell-associated tannase. The validity of the proposed model was verified, and the optimized medium was shown to produce maximum cell-associated tannase activity of 9.65 U/l, which is 93.8% higher than the activity in the basal medium, after 12 h at pH 5.0, 30 degrees C. The optimum medium consists of 38 g/l lactose, 50 g/l tryptose, and 2.8 g/l gallic acid.     

Optimization of L-asparaginase production from novel Enterobacter sp., by submerged fermentation using response surface methodology

The culture conditions and nutritional rations influencing the production of extra cellular antileukemic enzyme by novel Enterobacter aerogenes KCTC2190/MTCC111 were optimized in shake-flask culture. Process variables like pH, temperature, incubation time, carbon and nitrogen sources, inducer concentration, and inoculum size were taken into account. In the present study, finest enzyme activity achieved by traditional one variable at a time method was 7.6 IU/mL which was a 2.6-fold increase compared to the initial value. Further, the L-asparaginase production was optimized using response surface methodology, and validated experimental result at optimized process variables gave 18.35 IU/mL of L-asparaginase activity, which is 2.4-times higher than the traditional optimization approach. The study explored the E. aerogenes MTCC111 as a potent and potential bacterial source for high yield of antileukemic drug.     

Optimization of isoamyl acetate production by using immobilized lipase from Mucor miehei by response surface methodology

Immobilized lipase from Mucor miehei was employed for the esterification of isoamyl alcohol with acetic acid in n-heptane solvent. The important process variables studied were enzyme/substrate (E/S) ratio, alcohol (acid) concentration, and incubation period. Based on Box-Behnken design of experiments, a second order response function was developed. The percentage esterification increased with both E/S ratio and time and decreased with alcohol (acid) concentration. The model indicated optimum conditions for maximum esterification ranging from 20 to 99.6% in the alcohol (acid) concentration range of 0.031 to 0.3 M for a range of E/S ratios 8.33 to 50 g/mol, which were in good agreement with the experimental yields.     

Medium optimization for the production of recombinant nattokinase by Bacillus subtilis using response surface methodology

Nattokinase is a potent fibrinolytic enzyme with the potential for fighting cardiovascular diseases. Most recently, a new Bacillus subtilis/Escherichia coli (B. subtilis/E. coli) shuttle vector has been developed to achieve stable production of recombinant nattokinase in B. subtilis (Chen; et al. 2007, 23, 808-813). With this developed B. subtilis strain, the design of an optimum but cost-effective medium for high-level production of recombinant nattokinase was attempted by using response surface methodology. On the basis of the Plackett-Burman design, three critical medium components were selected. Subsequently, the optimum combination of selected factors was investigated by the Box-Behnken design. As a result, it gave the predicted maximum production of recombinant nattokinase with 71 500 CU/mL for shake-flask cultures when the concentrations of soybean hydrolysate, potassium phosphate, and calcium chloride in medium were at 6.100, 0.415, and 0.015%, respectively. This was further verified by a duplicated experiment. Moreover, the production scheme based on the optimum medium was scaled up in a fermenter. The batch fermentation of 3 L was carried out by controlling the condition at 37 degrees C and dissolved oxygen reaching 20% of air saturation level while the fermentation pH was initially set at 8.5. Without the need for controlling the broth pH, recombinant nattokinase production with a yield of 77 400 CU/mL (corresponding to 560 mg/L) could be obtained in the culture broth within 24 h. In particular, the recombinant B. subtilis strain was found fully stable at the end of fermentation when grown on the optimum medium. Overall, it indicates the success of this experimental design approach in formulating a simple and cost-effective medium, which provides the developed strain with sufficient nutrient supplements for stable and high-level production of recombinant nattokinase in a fermenter.     

Optimization of culture conditions for production of cis-epoxysuccinic acid hydrolase using response surface methodology

Response surface methodology, which allows for rapid identification of important factors and optimization of them to enhance enzyme production, was employed here to optimize culture conditions for the production of cis-epoxysuccinic acid hydrolase from Bordetella sp. strain 1–3. In the first step, a Plackett–Burman design was used to evaluate the effects of nine variables (yeast extract, cis-epoxysuccinic acid, KH₂PO₄, K₂HPO₄ · 3H₂O, MgSO₄ · 7H₂O, trace minerals solution, culture volume, initial pH and incubation time) on the enzyme production. Yeast extract, cis-epoxysuccinic acid and KH₂PO₄ had significant influences on cis-epoxysuccinic acid hydrolase production and their concentrations were further optimized using central composite design and response surface analysis. A combination of adjusting the concentration of yeast extract to 7.8 g/l, cis-epoxysuccinic acid to 9.8 g/l, and KH₂PO₄ to 1.12 g/l would favor maximum cis-epoxysuccinic acid hydrolase production. An enhancement of cis-epoxysuccinic acid hydrolase production from 5.6 U/ml to 9.27 U/ml was gained after optimization.