响应面法优化产低温脂肪酶工程菌Cl02的发酵条件

目的采用响应面法优化产低温脂肪酶工程菌Cl02的发酵条件。方法通过单因素试验考察培养基中酵母氮源含量、pH值、甘油含量和甲醇含量对酶活的影响,确定产酶的主要影响条件,在此基础上,根据Box-Benheken中心组合试验设计原理,采用三因素三水平的响应面分析法,综合考察各因子对低温脂肪酶酶活的影响,建立低温脂肪酶酶活的二次回归模型。结果培养基中酵母氮源含量、pH值和甲醇含量对酶活的影响显著。最适产低温脂肪酶条件为:酵母氮源含量7.3 g/L、pH 6.0、甲醇含量9.1 g/L,在此条件下,低温脂肪酶酶活可达42.25 IU/ml,比优化前(28.0 IU/ml)提高了50.9%。结论利用响应面法优化的发酵条件可显著提高工程菌的产酶能力,为规模化生产低温脂肪酶奠定了基础。     

表达乙型肝炎表面抗原重组酿酒酵母工程菌发酵培养基配方的优化

目的优化重组酿酒酵母工程菌发酵培养基的配方,提高其表达HBsAg的水平。方法采用正交试验法对培养基配方进行优化,并对最佳培养基配方进行验证。结果最佳培养基配方为:在每50ml基础培养基中添加50%的甘油0.9ml和50%的蔗糖0.4ml,维持pH值为5.0。与基础培养基相比,以此培养基培养的工程菌表达HBsAg的水平平均可提高26%。结论在发酵过程中适当补充甘油和蔗糖,可以提高重组酿酒酵母工程菌表达HBsAg的水平。     

响应面法优化醋酸菌的发酵产酸培养基

参考相关文献,选择10种醋酸菌发酵培养基成分,依次利用Plackett-Burman实验、最陡爬坡实验和Box-Behnken实验进行培养基成分优化,证实乙醇、葡萄糖和谷氨酸钠对醋酸产量影响显著.通过回归拟合方程(R2=97.68%),预测当培养基成分配比为乙醇9.8%、葡萄糖2.7%、谷氨酸钠0.4%、酵母粉1%、乙...     

棒状链霉菌产克拉维酸发酵培养基优化

以克拉维酸生产菌株Streptomyces clavuligerus K6为研究对象,考察了不同氮源对克拉维酸产量的影响,并通过二水平析因实验、快速爬坡实验和响应面法对发酵培养基碳源进行优化。确定最佳氮源为大豆蛋白,最佳发酵培养基碳、氮源配方为:甘油18.19 g·L~(-1)、糊精11.22 g·L~(-1)、三油酸甘油酯30.80 g·L~(-1)、大豆蛋白粉40 g·L~(-1)。在此发酵培养基配方下,克拉维酸产量可达5 220 mg·L~(-1),较出发配方提高58.1%。     

利用响应面分析法优化鸟苷发酵培养基

目的利用响应面分析法对鸟苷发酵培养基进行优化,提高鸟苷产量。方法采用响应面分析法对鸟苷发酵培养基的3种主要成分葡萄糖、酵母粉和豆饼水解液进行优化。用Design-Expert软件对实验数据进行多元回归分析,并建立3种因素与鸟苷产量之间的函数关系。用最终优化的配方进行3次验证试验。结果通过岭脊分析,获得培养基中3因素最佳浓度为:葡萄糖12.0%,酵母粉1.4%,豆饼水解液4.0%,发酵液鸟苷最高产量可达28.3g/L。3次验证试验所测得的鸟苷产量分别为28.14、28.32和27.86g/L,平均值为28.10g/L,与预测结果基本一致。结论利用响应面分析法可优化鸟苷发酵培养基,显著提高鸟苷的产量。     

夫西地酸发酵培养基优化

为了筛选夫西地酸高产培养基配方,提高夫西地酸发酵水平,以F17D2744为试验菌株,通过均匀设计方法,对不同碳氮源及微量元素等的配比实验,确定了最佳发酵培养基配方。试验结果表明:以蔗糖4.3%、糊精2%、酵母粉1%、蛋白胨3%、生物氮素0.59%、MgSO40.2%,KH2PO40.15%为发酵培养基配方,最终发酵效价较初始配方的发酵效价提高了234%。     

夫西地酸发酵培养基的初步优化

通过均匀设计法对夫西地酸的初始发酵培养基进行优化来获得更好的效价,以进一步降低企业生产成本,提高企业经济效益。经过多次试验,对所得最优解进行验证分析得到最优的发酵配方为:蔗糖4.3%、糊精2%、酵母粉1%、蛋白胨3%、生物氮素0.59%、硫酸镁0.2%、磷酸二氢钾0.15%;确定补料工艺为:96h补糖,补糖量1%,补加速率60mL/L。最终平均效价为1 105.6μg/mL,比最初配方得到的271.8μg/mL提高4倍。该试验验证了新型菌种的生产实用性。     

重组HIV-1 DNA疫苗工程菌中试发酵工艺的优化

目的优化重组HIV-1DNA疫苗工程菌的中试发酵工艺。方法优化质粒转化条件,制备工程菌Jm109/pVAX1-MEGp24种子液,连续传30代,检测其遗传稳定性;优化培养基成分、补料基质、补料方式和培养时间,确定最佳发酵参数;并应用最佳发酵参数,于50L发酵罐进行3批中试规模的发酵,考察在发酵培养过程中质粒的稳定性和超螺旋质粒的比例。结果工程菌在连续传代30次后,质粒拷贝数基本保持稳定;最佳发酵参数为:采用以甘油为碳源的培养基,以梯度恒速流加方式补料,培养时间13h;通过3批稳定发酵,最终可获得湿菌67.0～68.6g/L,质粒含量可达1.62～1.73mg/g菌,超螺旋质粒的比例达93%以上。结论已建立了稳定的重组HIV-1DNA疫苗工程菌中试发酵工艺,为进一步规模化生产奠定了基础。     

重组质粒DNA D-GPEi工程菌发酵工艺的优化

目的优化重组质粒D-GPEi工程菌发酵工艺条件。方法控制相关发酵参数,观察溶解氧浓度、比生长速率和培养时间对工程菌生长和质粒浓度的影响。结果发酵培养对数生长期溶解氧浓度控制在30%~50%。补料前比生长速率为0.5~0.7/h,6 h开始补料,比生长速率下降,12 h后比生长速率为0.1~0.2/h。连续发酵3批工程菌,21 h后收集菌体,得到菌体平均产量为89.4 g/L(A=52.6),质粒平均产量为359.8 mg/L。3批工程菌浓度和质粒拷贝数差异无显著意义。结论此发酵工艺得到工程菌浓度和质粒拷贝数较高,重复性好,适用于大规模工业化生产。     

东方拟无枝酸菌产万古霉素发酵工艺优化

采用东方拟无枝酸菌V19-07发酵产万古霉素,通过单因素实验对发酵工艺进行了优化,确定最优发酵工艺为:发酵温度28℃、发酵液pH值7.2、通气量1.0 vvm、溶解氧浓度10％.该工艺经50 L发酵罐验证,平均发酵效价达14117U·mL-1,产量较优化前提高18.8％,可用于规模化生产.     

响应面法优化以重组大肠杆菌发酵生产类人胶原蛋白过程

In order to improve the production of human-like collagen Ⅲ (HLC Ⅲ) by fed-batch culture of recom-binant Escherichia coli BL21, the Plackett-Burman and Box-Behnken design were applied to optimize the fermen-tation process parameters.Three variables (induction time, inoculum age and pH), which have significant effects on HLC Ⅲ production, were selected from eight variables by Plackett-Burman design.With the regression coefficient analysis in the Box-Behnken design, a relationship between HLC Ⅲ production and three significant factors was obtained, and the optimum levels of the three variables were as follows: induction time 3.2h, inoculum age 12.6 h and pH 6.7.The 3D response surface plots and 2D contour plots created by the Box-Behnken design showed that the interaction between induction time and pH and that between innoculum age and pH were significant.An aver-age 9.68 g·L~(-1)HLC Ⅲ production was attained in the validation experiment under optimized condition, which was 80% higher than the yield of 5.36 g·L~(-1) before optimization.     

Statistical optimization of process parameters for the production of vanillic acid by solid‐state fermentation of groundnut shell waste using response surface methodology

BACKGROUND: Vanillic acid is a flavoring agent and also serves as precursor for vanillin production. Culture medium and fermentation condition for the single step production of vanillic acid from Phanerochaete chrysosporium using lignocellulosic waste as a substrate under solid state fermentation (SSF) were optimized using response surface methodology. RESULTS: The process parameters were chosen by borrowing methodology, and L‐asparagine, pH and moisture content of the solid medium during SSF were identified as the most significant variables. The optimum value of selected variable and their mutual interactions were determined by response surface methodology. The result demonstrated that a yield of 73.58 mg vanillic acid g^?1 substate was predicted under optimum conditions (L‐asparagine 5.98 mmol L^?1 (2.37 mg g^?1 groundnut shell), pH of solid medium 4.51 and moisture content 74.83%). The predicted response was experimentally validated and resulted in a maximum vanillic acid yield of 73.69 mg g^?1 after 8 days of SSF. CONCLUSION: The optimization of fermentation variables resulted in a maximum 10‐fold increase in vanillic acid yield compared with that observed under sub‐optimal conditions (from 7.2 mg g^?1 to 73.69 mg g^?1). Copyright © 2011 Society of Chemical Industry     

生物表面活性剂生产Bacillus subtilis-1101发酵过程优化

应用中心组合试验设计和响应面分析方法对影响枯草芽孢杆菌Bacillus subtilis-1101产生表面活性剂的发酵过程进行优化。结果表明,枯草芽孢杆菌Bacillus subtilis-1101产生表面活性剂的最佳发酵条件为发酵温度29.1℃,初始pH值为4.9,装液量为56mL。在此条件下进行实验,结果最大排油圈为7.08cm,与模型预测值接近。说明响应面分析方法是优化表面活性剂生产的有力工具。     

短乳杆菌产胸苷磷酸化酶发酵培养基的优化

胸苷磷酸化酶在核苷类物质合成中具有重要作用,本研究以短乳杆菌为胸苷磷酸化酶生产菌种,对短乳杆菌产胸苷磷酸化酶发酵培养基进行优化。首先通过Plackett-Burman设计筛选出影响短乳杆菌产胸苷磷酸化酶的3个较为重要因素：发酵时间（P=0.030）、接种量（P=0.033）和葡萄糖浓度（P=0.019）。在此基础上采用最陡爬坡路径逼近最大响应区域,并利用响应面中心组合设计对影响显著因素进行优化,得到最适培养基组成成分和培养条件为：发酵初始pH 8.0,葡萄糖18 g/L,酵母膏15 g/L,NaCl 7.5 g/L,蛋白胨10 g/L,胸苷15 mmol/L,摇床转速110 r/min,发酵温度38 ℃,发酵时间10.57 h,接种量1.54%。在此优化条件下,短乳杆菌产胸苷磷酸化酶能力得到了很大提高,短乳杆菌胸苷磷酸化酶活从0.400 U/mg湿菌体提高到1.172 U/mg湿菌体,比优化前提高了2.93倍。蛋白质凝胶电泳分析显示经优化后每克湿菌体胸苷磷酸化酶的含量明显高于优化前。     

响应面分析优化霉酚酸发酵培养基

借助Design Expert 7.0软件,对霉酚酸发酵培养基的主要成分进行优化研究。首先采用全因子设计找出玉米浆和蔗糖为影响霉酚酸产量的主要因素,通过最陡爬坡法逼近最大响应区域,再利用中心组合设计及响应面分析法进行回归分析,求得两因素的最优水平：玉米浆98 g/L,蔗糖53 g/L。经验证,发酵培养基优化后菌株产量提高了38.2%。     

响应面法优化壳聚糖酶产生菌Mitsuaria sp.K1的产酶发酵条件

壳聚糖（chitosan）及其降解产物因具有优良的物理特性和生物活性而被广泛关注。通过平板透明圈初筛、摇瓶复筛方法,从青岛海岸土壤中分离筛选到1株产壳聚糖酶活性较高的细菌Mitsuaria sp.K1,并对其产酶发酵条件进行了单因素试验和响应面优化分析试验。结果表明：在最适培养基组成（1%粉末壳聚糖、0.5%硝酸钾、0.22%KH2PO4、0.1%Na2HPO4、0.15%KCl、0.05%MgSO4?7H2O）和最佳培养条件（培养温度25.2 ℃,培养时间25.4 h,起始pH值6.5,接种量3%,装液量100 mL/500 mL摇瓶,160 r/min）下,Mitsuaria sp.K1的发酵粗酶液最高酶活平均达11.56 U/mL,比优化前的2.17 U/mL提高了4.32倍。与前人研究结果相比,该菌发酵产酶温度降低了5～10 ℃,产酶周期缩短了23～47 h,因此具有工业发酵应用价值。     

Medium optimization for lipid production through co‐fermentation of glucose and xylose by the oleaginous yeast Lipomyces starkeyi

Co‐fermentation of lignocellulose‐based carbohydrates is a potential solution to improve the economics of microbial lipid production. In the present paper, experiments were performed to optimize the media composition for lipid production by the oleaginous yeast Lipomyces starkeyi AS 2.1560 through co‐fermentation of glucose and xylose (2 : 1 wt/wt). Statistical screening of nine media variables was performed by a Plackett–Burman design. Three factors, namely mixed sugar, yeast extract and FeSO₄, were found as significant components influencing cellular lipid accumulation. Further optimization was carried out using a Box–Behnken factorial design to study the effects of these three variables on lipid production. A mathematical model with the R² value at 96.66% was developed to show the effect of each medium composition and their interactions on the lipid production. The model estimated that a maximal lipid content of 61.0 wt‐% could be obtained when the concentrations of mixed sugar, yeast extract and FeSO₄ were at 73.3 g/L (glucose 48.9 g/L, xylose 24.4 g/L), 7.9 g/L and 4.0 mg/L, respectively. The predicted value was in good accordance with the experimental data of 61.5%. Compared with the initial media, the optimized media gave 1.59‐fold and 2.03‐fold increases for lipid content and lipid productivity, respectively.     

响应面优化木糖母液发酵产丁二酸

对产琥珀酸放线杆菌（Actinobacillus succinogenes）GXAS137发酵木糖母液产丁二酸的条件进行优化,探索利用废弃木糖母液合成高附加值丁二酸的可行性。首先通过Plackett-Burman实验设计确定影响丁二酸发酵的显著因子,然后采用最陡爬坡实验逼近各显著因子的最优区域,最后通过Box-Behnken实验设计确定各因子的最优水平。影响木糖母液发酵产丁二酸的显著因子及最优浓度分别为：木糖母液64.75g/L,玉米浆15.71g/L,碱式碳酸镁46.39g/L。在最优发酵培养条件下,丁二酸产量达到38.01g/L,比优化前提高了20.7%,与模型预测值（38.41g/L）基本一致。进一步利用2L发酵罐进行了放大试验,发酵72h丁二酸产量最高可达48.99g/L,较厌氧瓶发酵提高了28.9%,丁二酸得率为0.80g/g总糖。结果表明,采用低价的木糖母液作为底物,可为未来低成本、高效产业化生产丁二酸奠定坚实的基础。     

响应面法优化达托霉素发酵培养基

采用响应面法对玫瑰孢链霉菌发酵产达托霉素的培养基进行了优化。首先通过Placket-Burman设计法筛选出影响达托霉素产量的4个重要因素:初始pH值、葡萄糖、L-天冬氨酸(L-Asp)、硫酸钾。其中初始pH值的影响极为显著(p<0.01),因此,对初始pH值进行了单因素试验,得到最优初始pH值为8.6。在此基础上对其余3个因素用最陡爬坡路径逼近最大响应区域后,利用响应面中心组合设计对显著因素进行优化,得到最适培养基组成为:葡萄糖13.0 g/L、L-Asp 2.6 g/L、硫酸钾4.1 g/L。在此优化条件下,达托霉素产量达373.98 mg/L,与预测值(365.76 mg/L)非常接近,比优化前产量提高了2.25倍。