粘质沙雷氏菌G1利用玉米浆干粉和磷酸氢二铵生产2,3-丁二醇的研究

研究了几种工业氮源对粘质沙雷氏菌G1发酵生产2,3-丁二醇的影响,在确定玉米浆干粉为氮源的基础上,利用Plackett-Burman(PB)实验和响应面法(RSM)实验对玉米浆干粉和磷酸氢二铵[(NH4)2HPO4]的浓度进行了优化,确定优化培养基(g·L-1)为:蔗糖90,玉米浆干粉20.32,(NH4)2HPO47.21,NaAc 4,柠檬酸钠14,MgSO40.5,Fe-SO40.02,MnSO40.01。并以此优化培养基进行了摇瓶和分批补料发酵,结果表明,摇瓶发酵中,90g·L-1的蔗糖最终被转化成43.06g·L-1的2,3-丁二醇;分批补料发酵中,2,3-丁二醇浓度为128.28g·L-1,产率为2.67g·L-1·h-1,转化率为0.48g·g-1蔗糖。以玉米浆干粉和(NH4)2HPO4为氮源,2,3-丁二醇浓度较高,培养基的成本大幅降低,为工业化生产奠定了基础。     

枯草芽孢杆菌产芽孢发酵培养基的优化研究

在三角瓶培养条件下,采用单因素实验对枯草芽孢杆菌B53发酵培养基碳源、氮源进行了优化。在此基础上进行了正交实验,确定了菌株B53最佳的产孢发酵培养基为葡萄糖25g/L、大豆蛋白胨14g/L、玉米浆17g/L、NaCl 3g/L,MgSO_4 0.2g/L。在最优条件下发酵培养,芽孢浓度为9.6×10~9个/mL。     

酿酒酵母工程菌发酵产青蒿酸的工艺优化

通过酿酒酵母工程菌(Saccharomyces cerevisiae 1211)发酵制备青蒿酸,并通过单因素实验和响应面优化,考察了发酵温度、pH、半乳糖质量浓度、发酵碳源及发酵氮源等对青蒿酸发酵产量的影响。结果表明:在发酵温度30℃,发酵培养基初始pH=5.5,发酵培养基中蔗糖质量浓度91.8 g/L,半乳糖质量浓度10.1 g/L,硫酸铵质量浓度10.3 g/L,磷酸二氢钾质量浓度8.7 g/L的条件下,青蒿酸发酵产量可达(1529.7±12.6)mg/L,与未优化时的发酵产量相比,提升了67.1%。     

粪肠球菌HX-3-6产γ-氨基丁酸发酵条件的优化

目的优化粪肠球菌HX-3-6产γ-氨基丁酸(γ-Aminobutyric acid,GABA)的发酵条件,提高GABA的产量。方法通过单因素试验和正交试验对产GABA的粪肠球菌HX-3-6发酵条件进行优化,采用Plackett-Burman(PB)试验设计法筛选产GABA的培养基主要影响因素,应用Box-Behnken设计及响应面分析对影响发酵产GABA的主要培养基因素进行优化;用最终优化的配方进行3次验证试验。结果最佳发酵条件为底物浓度30 g/L,起始pH 5.5,发酵时间72 h,温度35℃;PB法筛选出了培养基中有显著效应的4个因素为MnSO4﹒H2O、NaCl、柠檬酸三胺和葡萄糖,经Box-Behnken设计及响应面分析,确定该4个主要影响因素的最佳浓度分别为0.100 785、0.224 459、0.215 355及12.335 95 g/L。3次验证GABA的产量平均可达16.027 g/L,比优化前(11.006 g/L)提高了45.62%。结论优化的粪肠球菌HX-3-6发酵条件和培养基可显著提高GABA产量,为其工业化生产奠定了基础。     

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。     

多杀菌素培养基配方的优化

采用正交实验法优化多杀菌素发酵培养基,得到优化配方：葡萄糖32 g/L、糊精60 g/L、酵母粉25 g/L、玉米浆15 g/L、KH2PO4 1.0 g/L、CaCO3 2.0 g/L、MgSO4 0.1 g/L,摇瓶效价较优化前提高了17.1%。     

玉米浆对大肠杆菌JH-B22发酵产L-丙氨酸的影响

为了进一步降低L-丙氨酸的生产成本,以大肠杆菌JH-B22为发酵菌株,以玉米浆为氮源、葡萄糖结晶废糖液为碳源进行L-丙氨酸的发酵。结果表明,发酵培养基中玉米浆添加量为20g·L~(-1)较为适宜。在玉米浆发酵培养基中进行L-丙氨酸的发酵,L-丙氨酸产量和糖酸转化率分别为54.30g·L~(-1)和90.5%;其L-丙氨酸产量略低于无机盐发酵培养基的56.12g·L~(-1)和LB发酵培养基的56.48g·L~(-1),但玉米浆发酵培养基具有配制更简单、无需额外添加无机盐或者成本较高的酵母粉等优点,可作为L-丙氨酸发酵生产的不错选择。     

响应面法优化胶原蛋白发酵培养基和培养条件

目的利用统计学方法对重组大肠杆菌发酵培养基进行优化,提高胶原蛋白产量。方法应用Plackett-Burman试验设计法和响应面法,对发酵培养基6种组分配比和2个初始发酵条件进行优化;用Design-Expert软件对实验数据进行多元回归分析,并建立3种主要因素(葡萄糖、混合氮源和K2HPO4)与胶原蛋白产量之间的函数关系。用最终优化的配方进行5次验证试验。结果培养基3个最佳浓度为:葡萄糖为14.69 g/L、混合氮源为15.04 g/L、K2HPO4为11.91 g/L,胶原蛋白表达率可达33.95%。5次验证试验所测得的胶原蛋白平均表达率为34.02%,与预测结果基本一致。结论优化的重组大肠杆菌发酵培养基可显著提高胶原蛋白产量。     

玉米浆水解液应用于谷氨酸发酵的研究

玉米浆含有丰富的营养物质,对谷氨酸的发酵生产影响较大。实际生产中,由于玉米浆的质量波动较大,谷氨酸发酵生产深受影响。为此,我们在发酵培养基中,添加一定量的玉米浆水解液进行发酵工艺优化实验。实验结果,谷氨酸产酸率达到了18.6 g·L-1,转化率达65.1%,谷氨酸产量提高了20%。发酵培养基组成确定为:淀粉水解糖55 g/L,玉米浆30 m L/L,玉米浆水解液10 g/L,豆粕水解液10 g/L,糖蜜55 g/L,K2HPO44 g/L,Mg SO4·7H2O 1.5 g/L,Mn SO4·7H2O 30 mg/L,Fe SO4·7H2O 30 mg/L,维生素B1350μg/L,维生素H 500μg/L,消泡剂0.1 m L/L。     

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。     

Modeling and optimization of cutinase production by recombinant Escherichia coli based on statistical experimental designs

Statistics-based experiment designs were used to optimize the culture medium (glucose, yeast extract, IPTG, tween-60, and CaCl₂) for cutinase production by recombinant Escherichia coli. A 2^5-1 fractional factorial design augmented with center points revealed that glucose, yeast extract, and IPTG were the most significant factors, whereas the other factors were not important within the levels tested. The method of steepest ascent was used to approach the proximity of optimum, followed by a central composite design to develop a response surface for culture condition optimization. The optimum culture medium for cutinase production was found to be: glucose 33. 92 g/L, yeast extract 30.92 g/L, and IPTG 0.76 g/L. A cutinase production of 145.27±1.5 U/mL, which was in agreement with the prediction, was observed in triplicate verification experiments. The results obtained here verified the effectiveness of the applied methodology and may be helpful for cutinase production on an industrial scale.     

响应面法优化费氏新萨托菌G5产β-甘露聚糖酶

采用响应面法对费氏新萨托菌G5产β-甘露聚糖酶发酵过程进行优化分析。在单因素实验的基础上选取8个因素,采用Plackett-Burman法进行筛选,确定NaCl和CaCl2浓度对β-甘露聚糖酶产量影响较大。用最速上升实验和中心组合实验进一步优化,利用Design-expert软件进行二次回归分析,得到最适宜发酵培养基配比为：瓜尔胶13.0 g/L,蛋白胨11.0 g/L,酵母粉5.0 g/L,MgSO4 1.0 g/L,KH2PO4 0.5 g/L,NaCl 0.6 g/L,CaCl2 0.6 g/L,pH值为4.0,此时酶产量为采用初始培养基发酵时酶产量的2.5倍。     

Determination of optimal conditions for ribonucleic acid production by <Emphasis Type="Italic">Candida tropicalis</Emphasis> no. 121

The experiments were based on multivariate statistical concepts, and response surface methodology (RSM) was applied to optimize the fermentation medium for the production of ribonucleic acid (RNA) by Candida tropicalis no. 121. The process involved the individual adjustment and optimization of various medium components at shake flask level. The two-level Plackett-Burman (PB) design was used to screen the medium components, which significantly influenced RNA production. Among seven variables, the concentrations of molasses, ZnSO₄, and H₃PO₄ were found to be the important factors that significantly affected RNA production (confidence levels above 95%). These factors were further optimized using a central composite design (CCD) and RSM. The optimum values for the critical components were as follows: molasses 47.21 g/L: ZnSO₄ 0.048 g/L; H₃PO₄ 1.19 g/L. Under optimal conditions, RNA production was 2.56 g/L, which was in excellent agreement with the predicted value (2.561 g/L), and led to a 2.1-fold increase compare with that using the original medium in RNA production.     

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。     

The role of environmental factors and medium composition on bacteriocin production by an aquaculture probiotic Enterococcus faecium MC13 isolated from fish intestine

The aim of this study was to optimize medium composition for higher yield of total viable cells and bacteriocin by Enterococcus faecium MC13. The factors such as peptone, meat extract, yeast extract, lactose, glycerol, tween 80, triammonium citrate and K₂HPO₄ were selected based on the Lactobacillus MRS medium composition. Two level factorial designs (FD) and steepest ascent path were performed to identify vital factors among the variables. Through the 2^?8 FD, peptone, yeast extract and lactose were found to be significant factors involved in the enhanced production of viable cells and bacteriocin. Therefore, these three foremost factors were further optimized by central composite design to achieve efficient yield. The optimum MRS composition was found to be peptone (40.0 g/L), meat extract (30.0 g/L), yeast extract (40.0 g/L), lactose (24.0 g/L), glycerol (5.8 g/L), Tween 80 (3.0 g/L), triammonium citrate (1.0 g/L), K₂HPO₄ (2.5 g/L), MgSO₄·7H₂O (0.10 g/L), MnSO₄·7H₂O (0.05 g/L) and dipotassium PO₄ (2.0 g/L). The optimized growth medium allowed higher amount of bacteriocin activity (36,100 AUml^?1) and total viable cells (14.22 LogCFUml^?1) production which were two-times higher than the commercial MRS medium.     

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。     

产L-乳酸的鼠李糖乳杆菌发酵培养基的优化

采用Plackett-Burman试验设计优化了鼠李糖乳杆菌产L-乳酸的发酵培养基,筛选出对L-乳酸产量有显著性影响的因素,即葡萄糖、柠檬酸氢二铵、乙酸钠、酵母膏,对这四个显著性因素,用中心组合试验优化得到最佳发酵培养基。结果表明:当培养基组成为:葡萄糖138.8 g/L、柠檬酸氢二铵2.19 g/L、乙酸钠6 g/L、酵母膏为7.56 g/L、蛋白胨0.50 g/L时,L-乳酸的产量为104.40 g/L。     

Optimization of medium for phenylalanine ammonia lyase production in <Emphasis Type="Italic">E. coli</Emphasis> using response surface methodology

A culture medium for phenylalanine ammonia lyase (PAL) production in E. coli was developed following preliminary studies by means of response surface methodology (RSM). The medium components having significant effect on the production were first identified by using a fractional factorial design. Then, central composite design (CCD) was used to optimize the medium constituents and explain the combined effects of four medium constituents: glucose, yeast extract, (NH₄)₂HPO₄ and MgSO₄. A quadratic model was found to fit the PAL production. CCD revealed that the optimum values of the test variables for PAL production were glucose 28.2 g/L, yeast extract 5.01 g/L, (NH₄)₂HPO₄ 7.02 g/L and MgSO₄ 1.5 g/L. PAL production of 62.85 U/g, which was in agreement with the prediction, was observed in the verification experiment. In comparison to the production of basal medium, 1.8-fold increase was obtained.     

餐厨垃圾同步糖化发酵生产燃料酒精的工艺

餐厨垃圾是指家庭、学校、食堂以及餐饮行业的食物废料和残余,是城市生活垃圾的重要组成部分。本文以学校食堂餐厨垃圾为原料,利用酿酒酵母对餐厨垃圾同步糖化发酵生产燃料酒精的工艺进行了研究。正交试验表明糖化酶和蛋白酶对酒精发酵影响显著,纤维素酶影响较小,糖化酶最适添加量为100U/g,蛋白酶最适添加量为150U/g,纤维素酶为100U/g,自然pH值(5.3)发酵,最佳的发酵周期是120h,最终酒精浓度达到54.6g/L。发酵过程无需添加其他营养物质,说明餐厨垃圾本身所含的营养物质即可以满足菌体生长的需要。     

利用响应面法优化出芽短梗霉As3．933产普鲁兰多糖发酵培养基

采用响应面分析法对出芽短梗霉As3．933产普鲁兰多糖的发酵培养基进行优化。首先利用Plackett-Bur-man实验筛选出影响普鲁兰多糖产量的主要因素为酵母膏、（NH4）zSO4和K2HPO4,再利用最陡爬坡实验逼近最大响应区域,最后通过Box-Behnken实验并运用Design-Expert8．0软件优化发酵培养基。确定优化培养基组成为：蔗糖62．5g·L-1,（NH4）2S040．67g·L-1,酵母膏2．84g·L-1,K2HPO47．12g·L-1,NaCl1．25g·L-1,MgS04·7H200．25g·L-1,pH值6．5,优化后的普鲁兰多糖产量达到22．29g·L-1,较初始液体发酵培养基（17．32g·L-1）提高了28．70％。