Effective extraction of elastase from Bacillus sp. fermentation broth using aqueous two-phase system

INTRODUCTION Aqueous two-phase system (ATPS) is widely used in biochemistry and biotechnology for purifica- tion of proteins (Diamond and Hsu, 1992; Balasubramaniam et al., 2003), enzymes (Pan et al., 2001; Bim and Franco, 2000), amino acids (Li et al., 2002), antibiotics (Yang et al., 1994; Guan et al., 1994), nucleic acids (Ohlsson et al., 1978; Kimura, 2000), aroma compounds (Marco et al., 2000) and lactic acid (Planas et al., 1998). ATPS is mainly composed of two-incompatible p…     

空气质量响应曲面模型在杭州市的应用

基于CMAQ模拟结果,统计、归纳出各种污染物排放控制因子与污染物浓度之间的函数关系,建立空气质量响应曲面模型RSM,通过留一法交叉验证和外部验证对模型进行可靠性验证。使用RSM,研究不同污染物的PM_2.5响应值,评估现有政策不变（Ⅰ）和政策收紧（Ⅱ）两种减排情景,发现NO_X、SO₂、NH₃和VOC对PM_2.5浓度体现明显的非线性响应特征,情景Ⅱ的减排效果明显好于情景Ⅰ,夏季的减排效果好于冬季。     

Effects of roller burnishing process parameters on surface roughness of A356/5%SIC composite using response surface methodology

In this study, a simple roller burnishing tool was made to operate burnishing processes on A356/5%SIC metal matrix composite fabricated by electromagnetic stir casting under different parameters. The effects of burnishing speed, burnishing force and number of burnishing passes on the surface roughness and tribological properties were measured. Scanning electron microscopy （SEM） graphs of the machined surface with PCD （insert-10） tool and roller burnished surface with tungsten carbide （WC） roller were taken into consider- ation to observe the surface finish of metal matrix composites. The mechanical properties （tensile strength, hardness, duc- tility） of A356/5%SIC metal matrix composites were studied for both unburnished samples and burnished samples. The results revealed that the roller burnished samples of A356/ 5%SIC led to the improvement in tensile strength, hardness and ductility. In order to find out the effects of roller bur- nishing process parameters on the surface roughness of A356/ 5%SIC metal matrix composite, response surface methodol- ogy （RSM） （Box-Behnken design） was used and a prediction model was developed relevant to average surface roughness using experimental data. In the range of process parameters, the result shows that roller burnishing speed increases, and surface roughness decreases, but on the other hand roller burnishing force and number of passes increase, and surface roughness increases. Optimum values of burnishing speed （1.5 m/s）, burnishing force （50 N） and number of passes （2） during roller burnishing of A356/5%SIC metal matrix com- posite to minimize the surface roughness （predicted 1.232 μm） have been found out. There was only 5.03% error in the experimental and modeled results of surface roughness.     

Influence of medium components on elastase production using crude sources by<Emphasis Type="Italic">Bacillus</Emphasis> sp. EL31410

A newly isolated strain EL31410, producing elastase (E. C3. 4. 4. 7) with high elastolytic activity was identified asBacillus sp. In the medium optimization, it was found that wheat bran and soybean flour hydrosate were the best crude carbon and nitrogen source for enzyme production, respectively. Addition of corn steep flour can affect the bacterium growth and elastase production. A fractional factorial design was applied to study the main factors that affect the enzyme production, and central composite experimental design and response surface methodology were adopted to derive a statistical model for the effect of wheat bran and soybean flour hydrosate on elastase production. The experimental results showed that wheat bran had positive effect but soybean flour hydrosate had negative effect, on enzyme production. An initial concentration of 3.4% (w/v) wheat bran and 9.4% (v/v) soybean flour hydrosate were found to be optimal for enzyme production in batch culture. The time course of elastase production in the optimized medium composition was also described. Project (No. 300024) supported by the Zhejiang Provincial Natural Science Foundation of China     

Optimization of succinic acid fermentation with Actinobacillus succinogenes by response surface methodology (RSM)

Succinic acid is considered as an important platform chemical. Succinic acid fermentation with Actinobacillus succinogenes strain BE-1 was optimized by central composite design (CCD) using a response surface methodology (RSM). The optimized production of succinic acid was predicted and the interactive effects between glucose, yeast extract, and magnesium carbonate were investigated. As a result, a model for predicting the concentration of succinic acid production was developed. The accuracy of the model was confirmed by the analysis of variance (ANOVA), and the validity was further proved by verification experiments showing that percentage errors between actual and predicted values varied from 3.02% to 6.38%. In addition, it was observed that the interactive effect between yeast extract and magnesium carbonate was statistically significant. In conclusion, RSM is an effective and useful method for optimizing the medium components and investigating the interactive effects, and can provide valuable information for succinic acid scale-up fermentation using A. succinogenes strain BE-1.     

Improvement of xylanase production by Aspergillus niger XY-1 using response surface methodology for optimizing the medium composition

Objective: To study the optimal medium composition for xylanase production by Aspergillus niger XY-1 in solid-state fermentation (SSF). Methods: Statistical methodology including the Plackett-Burman design (PBD) and the central composite design (CCD) was employed to investigate the individual crucial component of the medium that significantly affected the enzyme yield. Results: Firstly, NaNO3, yeast extract, urea, Na2CO3, MgSO4, peptone and (NH4)2SO4 were screened as the significant factors positively affecting the xylanase production by PBD. Secondly, by valuating the nitrogen sources effect, urea was proved to be the most effective and economic nitrogen source for xylanase production and used for further optimization.Finally, the CCD and response surface methodology (RSM) were applied to determine the optimal concentration of each sig-nificant variable, which included urea, Na2CO3 and MgSO4. Subsequently a second-order polynomial was determined by mul-tiple regression analysis. The optimum values of the critical components for maximum xylanase production were obtained as follows: x1 (urea)=0.163 (41.63 g/L), x2 (Na2CO3)=-1.68 (2.64 g/L), x3 (MGSO4)=1.338 (10.68 g/L) and the predicted xylanase value was 14374.6 U/g dry substrate. Using the optimized condition, xylanase production by Aspergillus niger XY-1 after 48 h fermentation reached 14637 U/g dry substrate with wheat bran in the shake flask. Conclusion: By using PBD and CCD, we obtained the optimal composition for xylanase production by Aspergillus niger XY-1 in SSF, and the results of no additional expensive medium and shortened fermentation time for higher xylanase production show the potential for industrial utilization.     

Regression-Based Artificial Neural Network Methodology In Response Surface Methodology

Response surface methodology (RSM) is an important tool for process parameter optimization, robust design and other quality improvement efforts. When the relationship between influential input variables and output response is very complex, it‘ s hard to find the real response surface using RSM. In recent years artificial neural network (ANN) has been used in RSM. But the classical ANN does not work well under the constraints of real applications. An algorithm of regression-based ANN(R-ANN) is proposed in this paper, which is a supplement to the classical ANN methodology. It makes network closer to the response surface, so that training time is reduced and robustness is strengthened. The procedure of improving ANN by regressions is described and the comparisons among R-ANN, RSM and classical ANN are computed graphically in three examples. Our research shows that the R-ANN methodology is a good supplement to the RSM and classical ANN methodology, which can yield lower standard error of prediction under conditions that the scope of experiment is rigidly restricted.     

嗜酸乳杆菌高密度发酵条件的研究

主要研究了嗜酸乳杆菌生长繁殖的环境条件(温度、接种量、起始pH等)和培养基组成对其影响.优化后确定了嗜酸乳杆菌的高密度培养条件为:起始pH值为5.8,培养温度为37℃,培养条件为亨-盖特厌氧培养,接种量为4%,培养时间为22 h.并且筛选了碳源,氮源及氮源的浓度.基配比为:2%乳清粉(w/v),1.5%牛肉膏(w/v),1.5%蛋白胨(w/v),0.058%硫酸镁(w/v),0.025%硫酸锰(w/v),0.22%乙酸钠(w/v),0.2%磷酸氢二钾(w/v).将该菌在此增菌培养基上培养,在37℃下培养22 h,菌数可达到1.17×1011cfu/mL.     

高校图书馆RSM体系的建构

引入工商企业界的CSM(用户满意管理),探索符合我国高校图书馆实际的RSM(读者满意管理)创新,并对高校图书馆为高等教育事业创新发展提供优质服务进行了探讨。     

Optimization of antioxidant activity by response surface methodology in hydrolysates of jellyfish (Rhopilema esculentum) umbrella collagen

To optimize the hydrolysis conditions to prepare hydrolysates of jellyfish umbrella collagen with the highest hydroxyl radical scavenging activity, collagen extracted from jellyfish umbrella was hydrolyzed with trypsin, and response surface methodology (RSM) was applied. The optimum conditions obtained from experiments were pH 7.75, temperature (T) 48.77 °C, and enzyme-to-substrate ratio ([E]/[S]) 3.50%. The analysis of variance in RSM showed that pH and [E]/[S] were important factors that significantly affected the process (P<0.05 and P<0.01, respectively). The hydrolysates of jellyfish umbrella collagen were fractionated by high performance liquid chromatography (HPLC), and three fractions (HF-1>3000 Da, 1000 Da<HF-2<3000 Da, and HF-3<1000 Da) were collected. The HF-2 fraction had the highest hydroxyl radical scavenging activity with the highest yield compared with the other two fractions. Furthermore, HF-2 also showed the strongest Cu²⁺-chelating ability and the best tyrosinase-inhibitory activity. Project (No. 2008BAD94B07) supported by the Hi-Tech Research and Development Program (863) of China