蛹虫草菌胞外多糖发酵及其发酵动力学

蛹虫草菌(Cordyceps militaris)胞外多糖(EPS)的发酵过程和发酵动力学进行了研究。基于Logitic方程和Luedeking-Piret方程,得到了描述发酵过程的动力学数学模型和模型参数,同时对实验数据与模型进行了验证比较。模型计算与实验结果拟合良好,模型正确地反应了蛹虫草菌胞外多糖的发酵过程及其动力学机制。     

放射形土壤杆菌Q9415发酵产胞外多糖动力学研究

放射形土壤杆菌Q9415产胞外多糖PS-9415的发酵为混合型发酵。应用logistic方程和L-P方程对PS-9415发酵过程的菌体生长,多糖合成以及底物消耗的动力学进行了讨论。研究发现：logistic方程和L-P方程建立的动力学模型较好地拟合了实验数据。     

放射形土壤杆菌Q9415发酵产胞外多糖动力学研究*

放射形土壤杆菌Q941 5产胞外多糖PS 941 5的发酵为混合型发酵。应用logistic方程和L P方程对PS 941 5发酵过程的菌体生长 ,多糖合成以及底物消耗的动力学进行了讨论。研究发现 :logistic方程和L P方程建立的动力学模型较好地拟合了实验数据。     

基于遗传算法的羊肚菌液体发酵动力学模型的建立

发酵动力学研究是实现发酵过程最优化控制及发酵过程放大的前提条件。本研究对羊肚菌液体深层发酵动力学进行了研究, 在Matlab软件平台上, 应用遗传算法对比了真菌生长较常用的Monod与Logistic方程在描述羊肚菌生长动力学时的优劣, 并对羊肚菌的生长、胞外多糖产生和基质消耗模型进行了参数估计。结果表明, Logistic方程与试验数据拟和情况更好, 并给出了羊肚菌液体深层发酵的动力学模型具体形式, 经验证, 模型的平均误差为5.8%。利用遗传算法选择羊肚菌动力学模型, 并进行参数估计与其他方法相比具有快速、搜索面广、接近全局最优解的特点, 在处理分批发酵动力学问题上具有不可比拟的优势, 发酵动力学模型的建立为发酵过程优化及放大奠定了基础。     

灵芝胞外多糖分批发酵动力学模型

利用分批发酵研究了灵芝(Ganoderma lucidum)胞外多糖的合成特性,结果表明Ganodermalucidum多糖合成和菌体生长呈部分生长关联型。菌体干重、胞外多糖分别达到15.56g·L-1、3.02g·L-1,胞外多糖对细胞干重得率系数(Yp/x)为0.19。根据分批发酵试验结果采用Logistic方程、Luedeking-Piret方程和类似Luedeking-Piret方程,得到了描述灵芝生长、胞外多糖以及葡萄糖底物消耗分批发酵动力学模型。同时在初始葡萄糖变化较大范围内,试验数据与模型预测值进行了比较拟合,平均相对误差小于5%,表现出很好的适用性。表明该动力学模型对指导灵芝胞外多糖的发酵生产具有实际意义。     

唾液链球菌嗜热亚种LCX2001胞外多糖分批发酵动力学*

对唾液链球菌嗜热亚种LCX2 0 0 1生物合成胞外多糖 (EPS)的动力学进行了研究 ,基于Logistic equation方程和Luedeking piret方程 ,得到了描述发酵过程的动力学模型和模型参数。该模型反映了细胞生长、乳酸生成、EPS生物合成和基质消耗之间的关系 ,模型的拟合结果与实验值吻合较好 ,误差小于 1 0 %。     

灵芝胞外多糖分批发酵动力学模型

利用分批发酵研究了灵芝（Ganoderma lucidum）胞外多糖的合成特性,结果表明Ganoderma lucidum多糖合成和菌体生长呈部分生长关联型。菌体干重、胞外多糖分别达到15.56g·L^-1<、3.02g·L^-1<,胞外多糖对细胞干重得率系数(Y_p/x）为0.19。根据分批发酵试验结果采用Logistic方程、Luedeking-Piret方程和类似Luedeking-Piret方程,得到了描述灵芝生长、胞外多糖以及葡萄糖底物消耗分批发酵动力学模型。同时在初始葡萄糖变化较大范围内,试验数据与模型预测值进行了比较拟合,平均相对误差小于5％,表现出很好的适用性。表明该动力学模型对指导灵芝胞外多糖的发酵生产具有实际意义。     

姬松茸N516产活性多糖的发酵动力学研究

目的 :研究姬松茸产活性多糖的发酵动力学。方法 :以 5L发酵罐分批发酵和基于Logistic和Luedeking&Piret方程。结果 :菌体生长和胞内活性多糖生成是同步的 ;发酵液流变特性参数稠度系数 (K)先增后降 ,而流变特性指数 (n)先降后升。得到描述分批发酵过程的动力学数学模型和模型参数 ,同时对实验数据与模型参数进行了验证比较 ,模型计算与实验结果较好拟合。结论 :产活性多糖属于偶联型以及模型可用于预测发酵过程     

唾液链球菌嗜热亚种LCX2001胞外多糖分批发醇动力学

对唾液链球菌嗜热亚种LCX2001生物合成胞外多糖（EPS）的动力学进行 了研究,基于Logistic-equation方程和Ｌuedeking-piret方程,得到了描述发酵过程的动力学模型和模型参数,该模型反映了细胞长长,乳酸生成,ＥＰＳ生物合成和基质消耗之间的关系,模型的拟合结果与实验值吻合较好,误差小于１０％。     

蛹虫草多糖和D-甘露醇深层发酵的非结构动力学模型

多糖和D-甘露醇是蛹虫草的重要药理活性成分。本文开展了蛹虫草在分批发酵过程中同时生产多糖和D-甘露醇的发酵动力学研究。利用Sigmoid函数构建了蛹虫草菌丝生长、糖基质消耗、多糖和D-甘露醇的非结构动力学模型,并根据Boltzmann方程拟合求解出各模型参数。结果显示,各模型的实测值和预测值拟合度较好。蛹虫草比生长速率在第1.0天达到最大值（μ_max）1.244d^-1;底物葡萄糖的比消耗速率在第0.6天达到最大值（q_{S, max}）2.163d^-1;多糖比合成速率在第2.0天达到最大值（q_{P, max}）51.852mg/(g·d);D-甘露醇比合成速率在第0.99天左右达到最大值（q_{D, max}）37.963mg/(g·d)。蛹虫草多糖的形成与菌丝细胞的生长呈现部分生长关联型,而D-甘露醇的形成与细胞生长呈现生长关联型关系。研究结果为利用分批发酵规模化同时发酵生产蛹虫草多糖和D-甘露醇提供了理论依据。     

Production of a Novel Extracellular Polysaccharide by Lactobacillus sake 0-1 and Characterization of the Polysaccharide

A novel exopolysaccharide (EPS) produced by Lactobacillus sake 0-1 (CBS 532.92) has been isolated and characterized. When the strain was grown on glucose, the produced EPS contained glucose and rhamnose in a molar ratio of 3:2 and the average molecular mass distribution (M(infm)) was determined at 6 x 10(sup6) Da. At a concentration of 1%, the 0-1 EPS had better viscosifying properties than xanthan gum when measured over a range of shear rates from 0 to 300 s(sup-1), while shear-thinning properties were comparable. Rheological data and anion-exchange chromatography suggested the presence of a negatively charged group in the EPS. Physiological parameters for optimal production of EPS were determined in batch fermentation experiments. Maximum EPS production was 1.40 g (middot) liter(sup-1), which was obtained when L. sake 0-1 had been grown anaerobically at 20(deg)C and pH 5.8. When cultured at lower temperatures, the EPS production per gram of biomass increased from 600 mg at 20(deg)C to 700 mg at 10(deg)C but the growth rate in the exponential phase decreased from 0.16 to 0.03 g (middot) liter(sup-1) (middot) h(sup-1). EPS production started in the early growth phase and stopped when the culture reached the stationary phase. Growing the 0-1 strain on different energy sources such as glucose, galactose, mannose, fructose, lactose, and sucrose did not alter the composition of the EPS produced.     

Production of extracellular polysaccharides by a Rhizobium species from root nodules of Cajanus cajan

The ability of the Rhizobium sp., isolated from the root nodules of the leguminous pulse yielding shrub Cajanus cajan, to produce extracellular polysaccharides (EPS) was checked. A large amount of EPS (1, 128 μg/ml) was produced by the bacteria in yeast extract mannitol medium. Growth and EPS production started simultaneously, but the production reached its maximum level in the stationary phase of growth at 28 h. The EPS production by this Rhizobium sp. was much higher than by many other strains from nodules of Cajanus cajan which took a much longer time to reach maximum EPS production than this strain. The maximum EPS production (2,561 μg/ml) was obtained when the medium was supplemented with mannitol (1%), cetyl pyridinium chloride (2 μg/ml) and KNO₃ (0.2%), in which the production was increased by 276% compared to the control. The EPS production rose in the period up to 65 h with increased mannitol concentration. The EPS contained arabinose, xylose and rhamnose monomers. The possible role of rhizobial EPS production in root nodule symbiosis is discussed.     

Metabolite profiles and growth characteristics of Rhizobium meliloti cultivated at different specific growth rates

Rhizobium meliloti (ATCC 55340) was grown at different specific growth rates in a chemostat apparatus. Metabolic products, relating to the Embden-Meyerhof-Parnas (EMP) pathway and the tricarboxylic acid (TCA) cycle, were measured and quantified to probe the influence of specific growth rate on the distribution of important metabolites. The detection of propionate in the fermentation broth implies that the imbalance of reducing equivalents of FADH(2) and NADH + H(+) resulted in a partially reductive operation of the TCA cycle. Additionally, experimental results show that the specific growth rate plays an essential role in modulating the biomass concentration, the specific substrate uptake rate, the cell length, the specific exopolysaccharide (EPS) production rate, the distribution of EPS molecular weight, and the profiles of carbohydrate and organic acid. The specific EPS production rate (varying from 13.3 to 111 mg EPS/g-DW/h) follows a growth-associated pattern at the specific growth rate ranging from 0.06 to 0.20 h(-1) and switches into non-growth-associated mode when the specific growth rate is over 0.20 h(-1).     

Production of exopolysaccharides by submerged culture of an enthomopathogenic fungus, Paecilomyces tenuipes C240 in stirred-tank and airlift reactors

The objective of this study was to investigate the effect of shearing effect on the production of exopolysaccharides (EPS) from an enthomopathogenic fungus, Paecilomyces tenuipes C240 in a stirred-tank reactor (STR) and in an airlift reactor (AR). The optimal agitation rate for the production of EPS in the STR was 150 rpm with the mycelial morphology of hairy pellets, where the final concentration and the specific production rate of EPS were 2.33 g l(-1) and 0.312 gg(-1) h(-1), respectively. However, the maximum concentration of biomass (21.06 g l(-1)) in the STR was obtained at a high agitation speed of 300 rpm. The specific production rate of EPS (0.456 gg(-1) h(-1)) in the AR was significantly higher than that achieved in the STR, in which the typical morphological form of mycelium was a loose clump. The three EPS groups in the STR (designated as STR-I, -II, and -III) and two groups of EPS in the AR (designated as AR-I and -II) were obtained from the culture filtrates by a gel filtration chromatography on Sepharose CL-6B. The molecular weights of STR-I, STR-II, STR-III, AR-I, and AR-II were determined to be 1,820, 25, 1.8, 1,160, and 6.7 kDa, respectively. An agitation rate of 150 rpm in the STR was selected as the optimal culture condition for maximum EPS production (2.33 g l(-1)), which was similar to the level achieved in the AR (2.30 g l(-1)). The carbohydrate composition in each EPS was quite different from each other: the major component was glucose (in STR-I, -III, and AR-I), mannose (in STR-II), and arabinose (in AR-II). In contrast, no significant difference in amino acid composition was observed.     

Exopolysaccharide production by Propionibacterium acidi-propionici on milk microfiltrate

AIMS: The aims of this work were to evaluate growth and exopolysaccharide (EPS) production properties of Propionibacterium acidi-propionici DSM 4900 on milk permeate. METHODS AND RESULTS: Anaerobic growth on milk permeate was only possible if supplemented with yeast extract (YE). Fermentation capacities of the strain were significantly improved by further increasing the supplemented YE. At 5 g l(-1) YE, consumption of 45 g l(-1) lactose to produce 9 g l(-1) biomass, 34 g l(-1) organic acids and 0.65 g l(-1) EPS was observed. From a kinetic point of view, EPS production occurred during the bacteria growth phase. At the excreted polysaccharide level, the medium showed shear-thinning behaviour with a relatively high apparent viscosity of up to 30 mPa.s (milli.Pascal.second) at a shear rate of 17 s(-1). CONCLUSION: EPS production by P. acidi-propionici DSM 4900 on milk permeate showed promising rheological behaviour of the milk-derived medium obtained, even at a low production level. SIGNIFICANCE AND IMPACT OF THE STUDY: A kinetic study on EPS production by a food-grade bacterium that could be used in situ in alimentation was carried out.     

Growth and exopolysaccharide production during free and immobilized cell chemostat culture of Lactobacillus rhamnosus RW-9595M

AIMS: Biomass and exopolysaccharide (EPS) production were studied during chemostat cultures in whey permeate medium with Lactobacillus rhamnosus RW-9595M-free cells and cells immobilized on solid porous supports (ImmobaSil). METHODS AND RESULTS: A continuous culture with free cells was conducted for 9 days at dilution rates (D) between 0.3 and 0.8 h(-1) in yeast extract (YE)/mineral supplemented whey permeate. Maximum EPS production (1808 mg l(-1)) and volumetric productivity (542.6 mg l(-1) h(-1)) were obtained for a low D of 0.3 h(-1). A continuous fermentation in a two-stage bioreactor system, composed of a first stage with immobilized cells and a second stage inoculated with free cells produced in the first reactor, was carried out for 32 days. The influence of YE concentration, temperature and dilution rate, and their interactions on biomass, EPS and lactic acid production was investigated. A statistically significant model was found only for lactic acid production. Marked cell morphological and physiological changes led to the formation of very large cell-containing aggregates and a low mean soluble EPS production (138 mg l(-1)). Aggregate volumetric productivity of the two-stage system varied between 5.7 and 49.5 g l(-1) h(-1) for different fermentation conditions and times. Aggregates contained a very high biomass concentration, estimated at 74% of aggregate dry weight by nitrogen analysis and 4.3 x 10(12) CFU g(-1) by a DNA extraction method and a high nonsoluble polysaccharide content (14.2%). At age 24 days, insoluble EPS concentration and volumetric productivity were 1250 mg l(-1) and 2240 mg l(-1) h(-1) respectively. The physiological changes were shown to be reversible when cells were incubated during three successive batch cultures. CONCLUSIONS: EPS production and volumetric productivity during continuous free-cell chemostat cultures with L. rhamnosus RW-9595M are among the highest values reported for lactobacilli in literature. Immobilization and continuous culture resulted in low soluble EPS production and large morphological and physiological changes of L. rhamnosus RW-9595M, with formation of macroscopical aggregates mainly composed of biomass and nonsoluble EPS. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study on continuous EPS production by immobilized LAB. Immobilization and culture time-induced cell aggregation and could be used to produce new synbiotic products with very high viable cell and EPS concentrations.     

Exopolysaccharide (EPS) biosynthesis by Lactobacillus sakei 0-1: production kinetics, enzyme activities and EPS yields

AIMS: To determine optimal exopolysaccharide (EPS) production conditions of the mesophilic lactic acid bacterium strain Lactobacillus sakei 0-1 and to detect possible links between EPS yields and the activity of relevant enzymes. METHODS AND RESULTS: Fermentation experiments at different temperatures using either glucose or lactose were carried out. EPS production took place during the exponential growth phase. Low temperatures, applying glucose as carbohydrate source, resulted in the best bacterial growth, the highest amounts of EPS and the highest specific EPS production. Activities of 10 important enzymes involved in the EPS biosynthesis and the energy formation of Lact. sakei 0-1 were measured. The obtained results revealed that there is a clear link for some enzymes with EPS biosynthesis. It was also demonstrated clearly that the presence of rhamnose in the EPS building blocks is due to high activities of the enzymes involved in the rhamnose synthetic branch. CONCLUSION: EPS production in Lact. sakei 0-1 is growth-associated and displays primary metabolite kinetics. Glucose as carbohydrate source and low temperatures enhance the EPS production. The enzymes involved in the biosynthesis of the activated sugar nucleotides play a major role in determining the monomeric composition of the synthesized EPS. SIGNIFICANCE AND IMPACT OF THE STUDY: The proposed results contribute to a better understanding of the physiological factors influencing EPS production and the key enzymes involved in EPS biosynthesis by Lact. sakei.     

Production of Extracellular Polysaccharide by a Rhizobium Species from Root Nodules of the Leguminous Tree Dalbergia lanceolaria

The ability of the Rhizobium D1 10 species, which was isolated from the root nodules of the leguminous forest tree Dalbergia lanceolaria, for the production of extracellular polysaccharides (EPS) was investigated. High amounts of EPS (765 μg/mL) were produced by the bacteria (Rhizobium D1 10) in yeast extract mannitol medium. Both growth and EPS production started simultaneously, but the EPS production was at its maximum in the stationary phase of growth at 32 h. The EPS production was maximal when the medium was supplemented with mannitol (2 %), thiamine hydrochloride (1 μg/mL) and KNO₃ (0.1 %), which was accompanied by a great increase in the production compared to the control. The EPS contained xylose, rhamnose, glucose, galactose and arabinose. The possible role of rhizobial EPS production in root nodule symbiosis is discussed.     

不同生长阶段青枯菌的高效离子交换色谱表征

采用高效离子交换色谱和激光光散射仪对不同生长阶段的青枯菌进行色谱表征。青枯菌经过色谱分离得到3个特征峰。通过对延滞期、对数生长期和稳定期青枯菌细胞浓度、发酵液pH值、细胞表面黏附的胞外酸性多糖(EPSⅠ)含量与青枯菌色谱行为的关系研究,发现在8h时,青枯菌运动能力强,细胞表面EPSⅠ少,吸附力弱,绝大多数菌体直接随流动相流出形成峰1;随着培养时间延长,细胞表面黏附的EPSⅠ量增多,与树脂吸附力增强,保留时间延长。因此分析3个色谱峰的形成与青枯菌的运动性、以及细胞表面EPSⅠ与阴离子交换树脂的吸附作用有关。并通过比较3个色谱峰青枯菌细胞表面EPSⅠ含量的差异和观察青枯菌经过4%甲醛固定处理后色谱行为的变化加以进一步验证。高效离子交换色谱为细菌完整细胞的研究提供了一种新的分析方法。     

Culture and exopolysaccharide production from sugarcane molasses by Gordonia polyisoprenivorans CCT 7137, isolated from contaminated groundwater in Brazil

Gordonia polyisoprenivorans CCT 7137 was isolated from groundwater contaminated with leachate in an old controlled landfill (São Paulo, Brazil), and cultured in GYM medium at different concentrations of sugarcane molasses (2%, 6%, and 10%). The strain growth was analyzed by monitoring the viable cell counts (c.f.u. mL^?1) and optical density and EPS production was evaluated at the end of the exponential phase and 24 h after it. The analysis of the viable cell counts showed that the medium that most favored bacterial growth was not the one that favored EPS production. The control medium (GYM) was the one that most favored the strain growth, at the maximum specific growth rate of 0.232 h^?1. Differences in bacterial growth when cultured at three different concentrations of molasses were not observed. Production of EPS, in all culture media used, began during the exponential phase and continued during the growth stationary phase. The highest total EPS production, after 24 h of stationary phase, was observed in 6% molasses medium (172.86 g L^?1) and 10% (139.47 g L^?1) and the specific total EPS production was higher in 10% molasses medium (39.03 × 10^?11 g c.f.u.^?1). After the exponential phase, in 2%, 6%, and 10% molasses media, a higher percentage of free exopolysaccharides (EPS) was observed, representing 88.4%, 62.4%, and 64.2% of the total, respectively. A different result was observed in pattern medium, which presented EPS made up of higher percentage of capsular EPS (66.4% of the total). This work is the first study on EPS production by G. polyisoprenivorans strain in GYM medium and in medium utilizing sugarcane molasses as the sole nutrient source and suggests its potential use for EPS production by G. polyisoprenivorans CCT 7137 aiming at application in biotechnological processes.