响应面法优化波赛链霉菌JMC 06001发酵培养基

采用响应面法对产生抑茵活性物质的波赛链霉茵(Streptomyces peucetius)茵株JMC06001的发酵培养基进行优化.首先采用Minnimum Run EquirepIicated Res IV设计对初始发酵培养基的8个营养因素进行筛选,获得影响产生抑茵活性物质的3个主要影响因素:葡萄糖、大豆粉和NaCl;然后用最陡爬坡实验快速逼近最大响应区域;最后,结合Box-Behnken设计及响应面分析,确定主要影响因素的最佳浓度,得出该菌株产抑茵活性物质的最优发酵培养基配方为:葡萄糖1.2%,麦芽糖0.7%,蛋白胨0.9%,大豆粉1.4%,NaCl 3.7%,CaCO3 0.1%,复合盐A液2.0%,复合盐B液0.1%,起始pH值7.0.用优化后的培养基发酵所得发酵液对敏感指示茵藤黄八叠球茵的抑茵圈直径达31.5 mm,与预测值的相对偏差仅为1.59%,与用初始发酵培养基发酵所得发酵液的抑茵效果(抑茵圈直径26.5 mm)相比提高了18.9%.     

响应面法优化纤维堆囊菌SoF5-76产埃博霉素B发酵培养基

以纤维堆囊菌SOF5-76为试验菌株,用响应面分析法对其产埃博霉素B的培养基进行优化,以提高埃博霉素B的产量。在单因素试验的基础上,利用Plackett-Burman筛选出对埃博霉素B产量有显著影响的3个因素为马铃薯淀粉、脱脂奶粉和无水氯化钙,在此基础上通过最陡爬坡试验逼近最佳响应面区域;再运用Box-Behnken试验设计和响应面分析法进行回归分析,确定重要因素的最优浓度。得到最佳发酵培养基为:马铃薯淀粉3.9 g/L、脱脂奶粉2.2 g/L、无水氯化钙1.3 g/L、葡萄糖1 g/L,豆饼粉1.5g/L,七水硫酸镁2.5 g/L,EDTA-Fe3+3 mL/L,微量元素(TE)0.5 mL/L,VB121 mL/L。在此最优条件下发酵埃博霉素B的产量为29.95 mg/L,与模型预测值接近,发酵产量比优化前提高了1.1倍。     

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。     

响应面法优化赖氨酸发酵培养基

运用Ｂｏｘ－Ｂｅｈｋｅｎ设计的响应面试验（Ｒｅｓｐｏｎｓｅ－ｓｕｒｆａｃｅ ｅｘｐｅｒｉｍｅｎｔ）对黄色短杆菌（Ｂｒｅｕｉｂａｃｔｅｒｉｕｍ ｆｌａｕｕｍ）ＦＢ４２进行赖氨酸发酵的培养基进行了优化。响应面法对７２小时分批发酵结果的优化分析表明：硫酸铵、豆饼水解液及玉米浆三种因素对赖氨醚发酵无明显的交互作用,在硫酸铵、豆饼水解液和玉米浆的浓度分别为５％、２％和３％时,发酵液中的产酸达到５４．０６ｇ／     

响应面法优化叶酸高产菌发酵培养基及发酵条件

采用响应面法对叶酸产生菌产朊假丝酵母Y2.12的发酵发酵培养基及发酵条件进行优化.首先,采用Plackett-Burman方法对影响发酵各因素的效应进行评价,筛选出有显著效应的3个因素：碳源乳糖、pH值和摇床转速;并通过Box-Benhnken设计和响应面分析法对这3个主要影响因素进行分析.结果表明,优化后这3个因素的最佳值为摇床转速196r/min、pH7和碳源乳糖含量为6.25%.采用优化后的发酵培养基及发酵条件进行摇瓶发酵,叶酸的含量可达23.14±0.13mg/L,取得了很好的优化效果.     

响应面设计优化绿僵菌固体发酵条件

【目的】为了提高绿僵菌分生孢子产量及孢子质量,应用响应面设计对金龟子绿僵菌菌株CY-1(Metarhizium anisopliae)进行固体发酵培养基的优化。【方法】单因素试验基础上,采用响应面试验设计方法优化培养基组分。【结果】添加了碳、氮营养的最佳固体发酵培养基为玉米粉:稻壳=8:2,料水比1:0.8,葡萄糖0.8%,硫酸铵2.5%,磷酸二氢钾0.8%;在固体培养基上的理论产孢量为7.45×10~9个/g;验证后实际为6.94×10~9个/g。【结论】运用响应面法对绿僵菌固体发酵的培养基成分进行优化,得到了绿僵菌孢子粉,为孢子粉进行地下害虫防治和制剂加工的研究奠定了基础。     

响应面法优化泰乐菌素发酵工艺的研究

以弗氏链霉菌(Streptomyces fradiae)为出发菌株,考察发酵工艺中摇瓶转速、接种量、发酵液初始pH,发酵温度4个因素对泰乐菌素含量的影响。在单因素试验的基础上,采用Box-Behnken响应面设计对各因素进行进一步优化,确定最终发酵条件为:摇瓶转速200r·min-1,接种量7.94%,发酵液初始pH 7.35,温度28.3℃,在此条件下发酵得到的泰乐菌素含量达到13 572μg·mL-1,比优化前提高了19.9%。     

利用响应面法优化ε-聚赖氨酸发酵培养基

采用响应面法对白色链霉菌ZC7发酵合成ε-聚赖氨酸的培养基进行优化研究。采用Plackett-Burman法对8个因素进行了筛选,结果表明,葡萄糖、酵母膏和硫酸铵的浓度对ε-聚赖氨酸产量影响较大。用最陡爬坡试验及Box-Behnken设计进一步优化,利用Design-Expert软件进行二次回归分析,得到各因素的最佳浓度为:葡萄糖37.22 g/L,酵母膏6.9 g/L,硫酸铵6.55 g/L。在此优化条件下ε-聚赖氨酸的产量达到8.11 g/L,较单因素试验最高值提高24.6%。     

响应面法优化青枯病生防菌Hrp~-菌株的发酵条件

对工程菌Hrp-菌株的摇瓶发酵条件进行优化。首先采用Plackett-Burman设计法对影响Hrp-菌株活菌量的6个因素进行评价,筛选出具有显著效应的2个因素(发酵温度和摇床转速),再用最陡爬坡试验法接近重要因子的最优水平,最后应用中心组合设计确定重要因子的最优水平。确定优化后的发酵条件:初始pH 8.75、装液量为100 mL(500 mL三角瓶)、接种量6.25%、接种龄10 h、温度20℃、摇瓶转速205 r/min。实验结果表明:采用优化后的发酵条件,Hrp-菌株的发酵液菌量由最初的3.30×1010个/mL提高至4.42×1010个/mL,增幅为133.9%。     

响应面法优化卑霉素发酵培养基的研究

在对卑霉素发酵培养基进行全因子试验和最陡爬坡试验后,运用Box-Behken设计的响应面试验(Response-surface experiment)对绿色产色链霉菌TN-29进行卑霉素发酵培养基的优化。响应面法对40h分批发酵结果的优化分析表明:豆饼粉、甘露醇及葡萄糖三种因素对卑霉素无明显的交互作用,在豆饼粉、甘露醇和葡萄糖的浓度分别为3.61%,2.99%,0.58%时,发酵液中效价值达到77.9U/ml的最大值。     

Application of response surface methodology in medium optimization for spore production of <Emphasis Type="Italic">Coniothyrium minitans</Emphasis> in solid-state fermentation

Summary Spore production of Coniothyrium minitans was optimized by using response surface methodology (RSM), which is a powerful mathematical approach widely applied in the optimization of fermentation process. In the first step of optimization, with Plackett–Burman design, soluble starch, urea and KH²PO⁴ were found to be the important factors affecting C. minitans spore production significantly. In the second step, a 2³ full factorial central composite design and RSM were applied to determine the optimal concentration of each significant variable. A second-order polynomial was determined by the multiple regression analysis of the experimental data. The optimum values for the critical components for the maximum were obtained as follows: soluble starch 0.643 (36.43 g. l⁻¹), urea −0.544 (3.91 g l⁻¹) and KH²PO⁴ 0.049 (1.02 g l⁻¹) with a predicted value of maximum spore production of 9.94 × 10⁹ spores/g IDM. Under the optimal conditions, the practical spore production was 1.04 × 10¹⁰ spores/g IDM. The determination coefficient (R²) was 0.923, which ensure an adequate credibility of the model.     

豆血红蛋白在毕赤酵母中的表达条件优化

【背景】豆血红蛋白可赋予素肉制品类似牛肉的红褐色质地,已被美国食品药物监督管理局批准作为人造素肉的着色剂,近年来受到广泛关注。【目的】优化毕赤酵母产豆血红蛋白的培养条件,提高毕赤酵母产豆血红蛋白的产量。【方法】首先通过单因素试验研究蛋白胨种类、大豆蛋白胨浓度、铁盐种类及血红素浓度在诱导阶段对毕赤酵母产豆血红蛋白的影响;然后通过Plackett-Burman试验设计筛选出对豆血红蛋白产量影响最大的3个因素,再通过最陡爬坡法确定3个因素的变化范围,对3个因素进行响应面分析;最后根据响应面结果进行摇瓶发酵和发酵罐高密度发酵。【结果】单因素试验发现：用4%大豆蛋白胨作为主要氮源、甲醇诱导浓度为1.5%、血红素浓度为5μmol/L时发酵效果较好,经过响应面优化后得到蛋白胨浓度为51.48 g/L、pH 5.66、培养基装液量35.84mL/250mL是最优发酵条件。在此优化条件下,LegH摇瓶发酵产量为0.191 mg/mL,与预测值(0.183 mg/mL)比较接近。采用5 L发酵罐进行高密度发酵,LegH产量最高达到0.384 mg/mL。【结论】优化了毕赤酵母表达豆血红蛋白的发酵条件,获得...     

一株高产洛伐他汀红曲霉的筛选与液态发酵条件优化

【背景】洛伐他汀(lovastatin)是红曲霉的次生代谢产物,是重要的临床用降血脂药物。在液态发酵条件下,红曲霉的洛伐他汀产量较低,难以满足工业化生产的要求。【目的】筛选获得一株高产洛伐他汀的红曲霉株,并通过优化液态发酵条件提高洛伐他汀的产量。【方法】从红曲米中筛选获得一株高产洛伐他汀的红曲霉株,依据形态学特征、生理生化特性及18S rRNA基因序列分析对分离菌株进行鉴定;通过响应面法对其产洛伐他汀的液态发酵条件进行优化。【结果】获得一株产洛伐他汀的紫红曲霉(Monascus purpureus M4),该菌在甘油57.80g/L、酵母浸粉5.52 g/L、接种量为6.90%条件下,洛伐他汀产量(173.60 mg/L)较优化前提高了4.8倍。【结论】菌株M4产洛伐他汀最优液态发酵条件的建立,为洛伐他汀的大规模生产及该菌株的工业化应用提供了技术支撑。     

黏质沙雷氏菌CS-1产灵菌红素的发酵条件优化

背景 灵菌红素(prodigiosin)是由黏质沙雷氏菌(Serratia marcescens)等微生物产生的一类三吡咯类次级代谢产物,具有较好的生物活性和应用潜力,但其发酵产量仍有待提高。目的 优化灵菌红素的发酵培养基成分和环境条件,实现其高效制备,以满足后续实验室生物活性研究产量需求。方法 采用多阶段优化策略对发酵工艺进行优化。首先通过单因素试验筛选适宜的碳源、氮源和无机盐;其次采用Plackett-Burman试验筛选显著影响因素,并结合最陡爬坡试验和Box-Behnken响应面法优化基础培养基组成及操作参数;在此基础上,进一步采用正交试验优化添加剂组合及环境条件。结果 获得的较优基础培养基组成为：葡萄糖5 g/L、蛋白胨8.2 g/L、NaCl 7.5 g/L;较优发酵操作参数为装液量150 mL/250 mL、接种量3.09%。添加剂较优组合为：吐温-80 5 g/L、L-脯氨酸1.0 g/L、甘氨酸1.5 g/L、组氨酸2.5 g/L。较优培养条件为：28 ℃、初始pH 7.0、180 r/min、发酵28 h。在上述条件下,灵菌红素产量达到193.4 mg/L,较原始发酵条件提高5.75倍。结论 通过单因素试验、Plackett-Burman试验、响应面分析及正交试验相结合的方法,建立了黏质沙雷氏菌CS-1产灵菌红素的较优摇瓶发酵条件。该结果为后续开展灵菌红素制备及应用研究提供了实验依据。     

Optimization and elucidation of interactions between ammonium, nitrate and phosphate inCentella asiatica cell culture using response surface methodology

The effects of macronutrients (NO₃ ⁻, NH₄ ⁺ and PO₄ ³⁻) on cell growth and triterpenoids production inCentella asiatica cell suspension cultures were analyzed using the Box-Behnken response surface model experimental design. In screening and optimization experiments, PO₄ ³⁻ as a single factor significantly influenced cell growth where increasing the phosphate level from 0.1 to 2.4 or 2.6 mM, elevated cell growth from 3.9 to 14–16 g/L. The optimum values predicted from the response surface model are 5.05 mM NH₄ ⁺, 15.0 mM NO₃ ⁻ and 2.6 mM PO₄ ³⁻, yielding 16.0 g/L cell dry weight with 99% fitness to the experimental data. While the NH₄ ⁺-NO₃ ⁻ interaction influenced cell growth positively in the optimization experiment, NH₄ ⁺ and NO₃ ⁻ as single factors; and interactions of NO₃ ⁻-PO₄ ³⁻, NH₄ ⁺-PO₄ ³⁻ and NH₄ ⁺-NO₃ ⁻ were all negative in the screening experiment. Cell growth and the final pH level were positively affected by PO₄ ³⁻, but negatively affected by NH₄ ⁺ and NH₄ ⁺-PO₄ ³⁻ interactions. The different effects of factors and their interactions on cell growth and final pH are influenced by a broad or narrow range of macronutrient concentrations. The productions of triterpenoids however were lower than 4 mg/g cell dry weight.     

响应面法优化L-谷氨酰胺发酵培养基的研究

目的:采用响应面法对L-谷氨酰胺发酵培养基成分进行优化,以提高L-谷氨酰胺发酵产量。方法:首先利用Plackett-Burman试验设计筛选出影响L-谷氨酰胺产量的主要因素葡萄糖、玉米浆和(NH4)2SO4,在此基础上采用最陡爬坡实验逼近最大响应区域,并利用中心组合设计及响应面分析法进行回归分析。结果:通过求解回归方程得到L-谷氨酰胺发酵培养基最佳浓度为葡萄糖100 g/L、玉米浆4.5 ml/L、(NH4)2SO437.2 g/L,L-谷氨酰胺产量理论最大值达41.0 g/L。结论:经模型验证,预测值与验证试验平均值接近,在优化条件下谷氨酰胺产量提高了37.6%。     

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%。     

Optimization of medium composition for actinomycin X2 production by <Emphasis Type="Italic">Streptomyces </Emphasis><Emphasis Type="Italic">spp</Emphasis> JAU4234 using response surface methodology

The effects of cultivation medium compositions including soybean meal, peptone, soybean oil and cornstarch for actinomycin X2 production by Streptomyces spp JAU4234 were accessed by using response surface methodology. The 2(4) full factorial designs and the paths of steepest ascent were effective in searching for the major factors of actinomycin X2 production. In this study, cornstarch and soybean oil showed negative effect on actinomycin X2 production based on the first-order regression coefficients derived from MINITAB software. Subsequently, a central composite design for optimization was further investigated. Preliminary studies showed that soybean meal and peptone were believed to be the major factors for actinomycin X2 production. Estimated optimum compositions for the production of actionmycin X2 were as follows (g/l): soybean meal 21.65 and peptone 9.41, and result in a maximum actionmycin X2 production of 617.4 mg/l. This value was closed to the 612 mg/l actionmycin X2 production from actual experimental observations. The yield of actionmycin X2 was increased by 36.9% by culturing the strain Streptomyces spp JAU4234 in the nutritionally optimized fermentation medium.     

响应面法优化短短芽胞杆菌ch2-22的发酵工艺

在摇瓶发酵条件下,优化提高短短芽胞杆菌ch2-22芽胞浓度和抑菌活性的发酵培养基和培养条件。首先在单因素试验基础上进行响应面设计对ch2-22的发酵培养基进行优化,然后使用单因素试验方法确定最佳发酵条件,得到优化发酵培养基为淀粉35.05 g/L,豆饼粉26.08 g/L,蔗糖10 g/L,鱼粉5 g/L,Na Cl 1 g/L,Mg SO40.3 g/L,(NH4)2SO43 g/L,Mn SO40.1 g/L,K2HPO43 g/L,酵母膏1 g/L,Ca CO32 g/L。培养条件为温度32℃,转速180 r/min,装液量100 m L/500 m L,接种量4%,初始pH为7.0,发酵时间45 h。优化后的芽胞数量达到8.1×109cfu/m L,与优化前的芽胞数量(1.6×109cfu/m L)相比,提高了3.7倍,优化后发酵液效价达到3 350 IU/m L,提高了109%,高于同类菌株的2 000 IU/m L。     

Statistical optimization of medium components for avilamycin production by <Emphasis Type="Italic">Streptomyces viridochromogenes</Emphasis> Tü57-1 using response surface methodology

A fermentation medium for avilamycin production by Streptomyces viridochromogenes Tü57-1 has been optimized. Important components and their concentrations were investigated using fractional factorial design and Box–Behnken Design. The results showed that soybean flour, soluble starch, MgSO₄·7H₂O and CaCl₂·2H₂O are important for avilamycin production. A polynomial model related to medium components and avilamycin yield had been established. A high coefficient of determination (R ² = 0.92) was obtained that indicated good agreement between the experimental and predicted values of avilamycin yield. Student’s T-test of each coefficient showed that all the linear and quadratic terms had significant effect (P > |T| < 0.05) on avilamycin yield. The significance of tested components was related to MgSO₄·7H₂O (0.37 g/L), CaCl₂·2H₂O (0.39 g/L), soybean flour (21.97 g/L) and soluble starch (37.22 g/L). The yield of avilamycin reached 88.33 ± 0.94 mg/L (p < 0.05) that was 2.8-fold the initial yield.