固定化德氏乳杆菌发酵乳酸及其微环境对细胞的影响

本文研究了以海藻酸载体的固定化德氏乳酸杆菌发酵乳酸的一些最佳条件及其微环境对于细胞增殖、细胞形态、α—氨基氮和无机磷的利用、热稳定性、严酸及操作稳定性等方面的影响。结果表明,在最佳条件下发酵液中乳酸含量达7.83％,转化率为81.1％,产率为1.63g/l.h,载体生产能力为8.16g乳酸/l载体.h。由于载体微环境的影响,固定化细胞的生长与形态和自由细胞有较大的不同,对α—氨基氮及无机磷的利用率降低,热稳定性及产酸量大幅度提高,并具有良好的操作稳定性。为适应工业生产的需要,研究了以麸皮水解渡与麸皮浸出液作营养源,以玉米淀粉水解液作底物的乳酸发酵。发现以麸皮水解液作营养源具有良好效果。     

戊糖乳杆菌发酵废甘油生产乳酸的条件优化在

对废甘油生产乳酸的发酵条件进行优化。得到乳酸发酵最佳条件：废甘油60 g/L,酵母粉2.8 g/L,蛋白胨4 g/L,硫酸铵5.82 g/L,乙酸钠6 g/L,磷酸氢二钾1.5 g/L,pH中和剂为氨水,控制pH值6.0,发酵温度34°C。在此条件下发酵70 h,乳酸产量达到51.43 g/L,不仅比优化前提高了约2%,而且减少了50%多的培养基费用,较大程度地减低培养基成本,为实现乳酸发酵产业化奠定基础。这是首次报道戊糖乳杆菌发酵废甘油生产乳酸。     

响应面法优化乳杆菌发酵生产奶味香精的工艺研究

主要研究了利用响应面法优化保加利亚乳杆菌发酵生产奶味香精的工艺条件。选择底物浓度、菌种接种量、发酵温度、发酵时间为考察因素,利用Design-Expert软件进行设计和分析,确定了发酵产香的最佳工艺条件为:底物浓度21%、菌种接种量16%、发酵温度38℃、发酵时间21 h。     

植物乳杆菌XL-02发酵产γ-氨基丁酸条件的优化

为提高γ-氨基丁酸(GABA)产量,对植物乳杆菌XL-02进行发酵条件的优化.首先,采用单因素方法对底物添加量、发酵温度、发酵时间、接种量和pH值等因素进行考察,在此基础上进行4因素3水平的响应面实验优化.结果表明:在L谷氨酸钠浓度为2.0 g/L,发酵温度30℃,接种量11％和pH值6.0的条件下,发酵60 h,γ-...     

二次正交旋转组合设计优化罗伊乳杆菌发酵培养基

目的优化猪源罗伊乳杆菌的发酵培养基4个组分的配比。方法应用SASV8.0统计分析软件中的二次正交旋转组合设计方案进行试验设计,并通过响应面法分析各因素对响应值的效应关系。结果优化后培养基各组分比例为:大豆蛋白胨5%,葡萄糖1%,酵母浸粉1.7%,低聚糖0.3%。应用此配比进行了3次重复发酵试验,A620的平均值为1.073,与预测值(1.092)基本相符。结论二次正交旋转组合设计结合响应面法可用于发酵培养基组分的优化和分析。     

干酪乳杆菌发酵罗非鱼下脚料制备L-乳酸研究

本实验以罗非鱼(Oreochromis spp)下脚料的水抽提液为原料,研究干酪乳杆菌(Lactobacillus casei)在不同氮源中的生长情况和产L-乳酸量,探讨罗非鱼水抽提液在干酪乳杆菌发酵L-乳酸中代替蛋白胨作为氮源的可能性。结果表明:干酪乳杆菌在水抽提液中的生长状况与在蛋白胨发酵液中相似,在42℃发酵14 h后,以水抽提液为氮源的发酵液中L-乳酸净产量为925.8 mg/100g,仅比在蛋白胨氮源的少12 mg/100g。     

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。     

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。     

乳酸菌发酵甘蔗糖蜜产乳酸的培养条件优化

乳酸是有广泛应用的有机酸,廉价原料是降低乳酸生产成本的重要因素。以实验室前期筛选的植物乳杆菌sy4为出发菌株研究了利用甘蔗糖蜜生产乳酸的培养条件。在确定发酵温度和初始pH后,通过Plackett-Burman实验和中心复合实验优化乳酸发酵条件。结果表明：植物乳杆菌sy4乳酸发酵的最适条件为温度32℃、60h、初始pH 6.5;Plackett-Burman实验表明酵母提取物、糖蜜和碳酸钙是影响乳酸产量的主要因素;中心复合实验得到三因素的最优组合为：酵母提取物13.19g/L、糖蜜476.63g/L和碳酸钙134.82g/L,确定了最适发酵培养基。在此条件下乳酸产量为（145.53±1.24） g/L,与模型预测值147.23g/L接近。本研究为以甘蔗糖蜜为原料生产乳酸提供了技术支持。     

大豆秸秆酶水解及L-乳酸发酵

用纤维素酶对氨预处理后的大豆秸秆进行酶水解,利用纤维素酶的作用使纤维素、半纤维素水解为可溶性糖,继而研究了用干酪乳杆菌及清酒乳杆菌进行L-乳酸发酵,通过微生物发酵将生成的可溶性糖转化为用于生产具有可生物降解性的聚乳酸塑料的原料乳酸,实现可再生资源的充分利用.结果表明,实验条件下,5%的大豆秸秆经酶水解后,还原糖浓度为242.25 mg•g^-1,纤维素糖化率为51.22%.清酒乳杆菌、干酪乳杆菌及该两种混合菌种发酵酶解液所得L-乳酸的转化率分别为 48.27%、56.42%和71.05%.     

The stability of lactic acid production in resting suspensions of Lactobacillus delbrueckii

The stability of resting glycolytic activity (RGA) i.e. lactic acid production from glucose, in non-growing suspensions of Lactobacillus delbrueckii was examined. Non-growing conditions were imposed by removal of essential nutrients or by the addition of antibiotics. The stability of RGA was reduced in fast-grown chemostat cells. Mg²⁺, reducing agents, the rigorous exclusion of oxygen and a constant supply of energy source gave a five-fold increase in RGA and a 25-fold increase in stability in cells deprived of a nitrogen source. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) activity severely limited RGA after about 8 h of incubation. No evidence of excessive ATP accumulation or ADP depletion was found. Addition of chloramphenicol, actinomycin D or mitomycin C decreased RGA stability over a 24-h period. Addition of yeast extract and tryptone to a mixture containing phosphate, Mg²⁺, thioglycollate and chloramphenicol gave a three-fold increase in RGA which lasted for about 1 h. Mg²⁺ is believed to stimulate the glycolytic sequence directly and oxygen probably inhibits GAPDH by interaction with the -SH groups of the enzyme. Tryptone and yeast extract probably stimulate pyruvate kinase activity, and this enzyme is also absolutely dependent on fructose-1, 6-diphosphate for activity.     

Production of lactic acid from beet molasses by calcium alginate immobilized Lactobacillus delbrueckii IFO 3202

Lactic acid was produced from pretreated beet molasses by the homofermentative organism Lactobacillus delbrueckii subsp delbrueckii IFO 3202 entrapped in calcium alginate gel using batch, repeated batch and continuous fermentation systems. In batch fermentation studies successful results were obtained with 2.0–2.4 mm diameter beads prepared from 2% sodium alginate solution. The highest effective yield (82.0%) and conversion yield (90.0%) were obtained from substrate concentrations of 52.1 and 78.2 g dm⁻³ respectively. The gel beads produced lactic acid for 14 consecutive batch fermentations without marked activity loss and deformation. In the continuous fermentation, the highest lactic acid (4.22%) was obtained at a dilution rate of 0.1 h⁻¹ while the highest productivity (13.92 g dm⁻³ h⁻¹) was obtained at a dilution rate of 0.4 h⁻¹. © 1999 Society of Chemical Industry     

Kinetics of lactic acid fermentation on glucose and corn by Lactobacillus amylophilus

The biosynthesis of ammonium lactate, a product of lactic acid fermentation was studied from corn and glucose at five different pH values of 5.4 to 7.8. In the glucose fermentations, a 100% conversion of substrate was obtained resulting in a maximum lactic acid production yield of 93.2%. The optimum pH for the maximum volumetric rate of lactic acid biosynthesis (1.56 g dm^?3 h^?1) was between 6.0 and 6.5. The corn fermentations were slower than the glucose fermentations with a resulting lactic acid yield of 67.5%. Hydrolysis of corn by enzymatic or chemical methods as well as the use of ammonium hydroxide for pH control increased both the final concentration and the rates of lactic acid production. An enhanced yield of more than 90% was finally obtained in the corn fermentations. A logistic model adequately described the kinetics of biomass growth, lactic acid production and sugar utilization in the glucose fermentations at different pH values. The dynamics of lactic acid formation in the corn fermentations were also successfully described by the developed model. The dependence of the model parameters on pH was investigated.     

Unstructured mathematical model for biomass,lactic acid and bacteriocin production by lactic acid bacteria in batch fermentation

BACKGROUND: A simple macroscopical model was proposed to describe the fermentation kinetics of growth, bacteriocins and lactic acid production by Lactococcus lactis and Pediococcus acidilactici in a batch system. The equations used were: the logistic reparametrized for growth, the Luedeking–Piret model for bacteriocin production, the maintenance energy model for glucose consumption; and the homofermentative balance equation for lactic acid formation. RESULTS: In all the cultures, the mathematical models, consistents and robusts, adjusted, perfectly, the experimental kinetic profiles. Also, the corresponding kinetic parameters were significant, so much biological as statistically. CONCLUSIONS: The group of integrated equations used, besides showing high accuracy in predicting the studied bioproductions, established a useful tool for the control of lactic acid bacteria kinetics in bioreactors in terms of its statistical consistency. Copyright © 2007 Society of Chemical Industry     

重组柠檬酸杆菌合成紫色杆菌素的工艺条件优化

以L-色氨酸为前体物,对弗氏柠檬酸杆菌(Citrobacter freundii)基因工程菌株生产紫色杆菌素的工艺条件进行了优化。通过单因素实验研究了培养基种类、培养温度、碳源、溶氧、种子液的状态、接种量、诱导时机、诱导剂的浓度及初始pH对细胞生长和紫色杆菌素产量的影响,通过正交实验研究了这些因素中可能存在交互作用的4个因素(种子液OD660、接种量、诱导时机、诱导剂浓度)的影响主次,确定了这些因素、水平的最佳搭配方案。结果表明,重组柠檬酸杆菌合成紫色杆菌素的最优培养条件为:种子液培养至OD660=3.6时,以3%的接种量接种于以甘油为碳源、初始pH为6.5的磷酸盐发酵培养基E2(25μg.ml-1卡那霉素),先在37℃、200r.min-1下培养至OD660=1.4,然后加入0.5μl.ml-1的诱导剂正辛烷,同时转入20℃,在150r.min-1下诱导培养31h。在此最优条件下,紫色杆菌素粗提物(紫色杆菌素及脱氧紫色杆菌素的混合物)产量可达1.809g.L-1,比优化前(0.514g.L-1)提高了252%,是目前国际上其他研究小组报道的最高摇瓶产量(0.43g.L-1)的4.2倍。质粒稳定性实验结果表明重组柠檬酸杆菌在抗生素选择压力条件下具有良好的遗传稳定性。     

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。     

Modeling and simulation of tanks-in-series airlift bioreactors for production of lactic acid by fermentation

A tanks-in-series model was applied for mathematical modeling of the unsteady state performance of 70 and 100 liters airlift bioreactor for the production of lactic acid by fermentation. A set of first-order differential equations for the material balances of micro-organism, substrate, product, and dissolved oxygen around hypothetically well mixed stages was solved simultaneously utilizing computer program in MATLAB. The kinetic model utilized considered the effect of two substrates (glucose and dissolved oxygen) on the growth rate. The effect of air velocity on the lactic acid production was investigated. Results of this model have been validated with experimental data.     

聚乙烯醇-海藻酸钙作为德氏乳酸杆菌包埋剂的研究

以德氏乳酸杆菌为考察对象,应用响应面分析法(RSM)对聚乙烯醇-海藻酸钙凝胶配比进行优化研究.结果表明,制备PVA-海藻酸钙复合载体的最佳条件是PVA、海藻酸钠和氯化钙的质量分数分别为6.64%、1.29%和1.97%,固化交联时间为6.48 h,预测乳酸发酵72 h的产量为112.79 g/L.经过120 d,40批次的发酵验证,结果与预测值相符,且乳酸发酵72 h产量比游离细胞发酵平均提高了56%.     

Production of L‐lactic acid by Lactobacillus delbrueckii in chemostat culture using an ion exchange resins system

L ‐Lactic acid production from beet molasses by Lactobacillus delbrueckii CECT 286 in a continuous fermenter‐ion exchange resins system has been investigated and compared with that observed in a conventional chemostat. The principle of this method is to remove the lactate during the course of fermentation as it is formed by adsorption to an anion exchange resin (Amberlite IRA‐420) in the carbonate form and to overcome its inhibitory effects on lactic acid bacteria by maintaining low lactate concentrations in the medium. Ammonium lactate was formed by percolating ammonium carbonate solution through this resin and it was converted to lactic acid by treatment with a cation exchange resin (Amberlite IR‐120) in hydrogen form. Compared with a conventional chemostat, this fermentation–ion exchange resins system enhanced the fermentation, controlled the pH, and showed the remarkable effect of increasing the yields of lactic acid from sucrose and biomass from sucrose due to the complete utilization of sucrose. © 1999 Society of Chemical Industry