木糖醇发酵状态估算、过程预测与优化

运用广义对数方程拟合不同初始木糖浓度(4.73～94.71g/L)下的木糖醇发酵过程,借助于均匀设计综合考察构建神经网络时第一、二隐层单元数、学习速度、初始权矩阵对模拟结果的影响及不同的初始状态变量对木糖醇发酵过程的影响,建立了一个能够很好地用于不同初糖浓度下木糖醇发酵状态估算和过程预测的四层6-7-6-3拓扑构型的神经网络模型,提出了对改善木糖醇发酵有指导意义的优化方案。     

木糖醇发酵状态估算,过程预测与优化

运用广义对数方程拟合不同初始木糖浓度（４７３～９４７１ｇ／Ｌ）下的木糖醇发酵过程,借助于均匀设计综合考察构建神经网络时第一、二隐层单元数、学习速度、初始权矩阵对模拟结果的影响及不同的初始状态变量对木糖醇发酵过程的影响,建立了一个能够很好地用于不同初糖浓度下木糖醇发酵状态估算和过程预测的四层６７６３拓扑构型的神经网络模型,提出了对改善木糖醇发酵有指导意义的优化方案。     

谷氨酸流加发酵过程的神经网络优化研究

采用神经网络的方法,将模拟网络与优化网络结合组成双网系统,用于谷氨酸流加生产过程的模拟与优化分析。模拟网络中设置瓶颈结构．以加强网络的数据过滤与压缩能力,滤除工业数据中的噪音。利用训练算法本身,优化网络方便地实现了单变量、多变量优化,获得令人满意的结果。     

谷氨酸发酵过程葡萄糖自动流加系统

补料（流加葡萄糖）操作是谷氨酸发酵过程最重要的操作之一,工业上有各种各样的补料操作方法。控制葡萄糖浓度于一个较为平稳且适中的水平有利于提高谷氨酸发酵的性能指标。通过在线计量谷氨酸发酵中的氨水耗量并据此在线推定发酵液中的葡萄糖浓度,构建了一个谷氨酸发酵自动在线补料系统。使用该控制系统,谷氨酸发酵过程的葡萄糖浓度可以控制在任意水平,平稳、无波动的谷氨酸发酵可以得到实现。     

谷氨酸棒杆菌一步法发酵糖质原料生产γ-聚谷氨酸

γ-聚谷氨酸在食品、化妆品、生物医药等领域具有广泛的应用,目前主要的生产菌株是谷氨酸依赖型菌株,在生产过程中需要添加谷氨酸作为前体,因而生产γ-聚谷氨酸的成本较高。文中主要研究从糖质原料一步法发酵合成γ-聚谷氨酸的生产工艺。首先,从产γ-聚谷氨酸的菌株枯草芽孢杆菌中克隆γ-聚谷氨酸合成酶的基因簇pgs BCA,在谷氨酸棒杆菌模式菌株ATCC13032中进行诱导型和组成型表达,结果显示,仅诱导型表达菌株可以积累γ-聚谷氨酸,产量为1.43 g/L。进一步对诱导条件进行优化,确定诱导时间为2 h,IPTG浓度为0.8 mmol/L,γ-聚谷氨酸产量为1.98g/L。在此基础上,在一株高产谷氨酸的谷氨酸棒杆菌F343中外源表达pgs BCA,对重组菌进行发酵,结果表明,在摇瓶发酵中γ-聚谷氨酸产量达到10.23g/L,在5L发酵罐中产量达到20.08g/L;继而对γ-聚谷氨酸进行分子量测定,结果显示,产自F343重组菌的γ-聚谷氨酸的重均分子量比产自枯草芽孢杆菌的提高34.77%。文中构建了一步法发酵糖质原料生产γ-聚谷氨酸的新途径,同时为开发其潜在应用奠定了基础。     

谷氨酸棒杆菌发酵过程中亮氨酸浓度近红外模型的建立

目的:建立谷氨酸棒杆菌发酵过程中亮氨酸浓度近红外模型,为实现谷氨酸棒杆菌发酵生产亮氨酸的发酵过程自动化控制提供理论基础和实践依据。方法:首先在5 L发酵罐中进行亮氨酸发酵,每隔一段时间采集发酵液样品,用高效液相色谱精确分析各样品中的亮氨酸实际浓度,再利用近红外分析仪和相关软件,对各样品进行近红外光谱扫描分析,并通过近红外光谱分析软件进行数据处理,建立谷氨酸棒杆菌发酵过程中亮氨酸浓度的近红外预测模型,最后通过外部检验方法检验模型的准确性。结果:结合偏最小二乘法,在波长为9043.3~7489.1 cm-1、减去一条直线作为光谱预处理的条件下,获得谷氨酸棒杆菌发酵过程中亮氨酸浓度最优近红外预测模型。该模型交叉验证误差均方根(RMSECV)、决定系数(R2)以及剩余预测偏差(RPD)分别为1.29 g/L、0.977和4.55。结论:经过验证,该模型的准确性和可靠性较强,实际值与预测值之间的误差较小,能够较好地检测发酵过程中的亮氨酸浓度。     

谷氨酸产生菌FM820-7菌株一次性中糖发酵的研究

自木下等人首先报导谷氨酸发酵工程的研究工作以来,已有很大发展。采用以糖蜜为原料的低糖流加技术进行生产,能得到较好的产酸率。但是由于各种因素的限制,绝大多数厂家还不能采用这一技术。为提高谷氨酸的发酵水平,目前我国的生产厂都正从低糖发酵向中糖发酵过     

具有时滞的双向联想记忆神经网络的全局渐近稳定性

双向联想记忆模型是两层异联想网络,本文讨论了具有轴突信号传输时滞的双向联想记忆神经网络的全局渐近稳定性,得出了保证神经网络平衡点稳定的几个充分条件,所得到的结论对于具有时滞的连续双向联想记忆神经网络的设计和应用都是很有意义的。     

混菌发酵生产γ-聚谷氨酸的工艺条件优化

采用谷氨酸棒杆菌S9114和枯草芽胞杆菌NTG-4在10 L自控发酵罐上进行混菌发酵,探索混菌发酵生产γ-聚谷氨酸的可行性并进行工艺优化。结果表明:温度、接种量、pH及溶氧对聚谷氨酸发酵有较大影响,发酵前期维持32℃,6 h提温至37℃变温控制,谷氨酸棒杆菌和枯草芽胞杆菌接种量分别为5%和0.5%,pH 7.0,溶氧20%最有利于γ-聚谷氨酸发酵,在此条件下发酵32 hγ-聚谷氨酸最高产量为38.3 g/L。     

基于pH的反馈补料方法在谷氨酸发酵中的应用

为简化谷氨酸发酵补料工艺,提出了一种新型的基于pH的补料方式。考察谷氨酸发酵过程中氨消耗量 (x) 和糖消耗量 (y) 发现,两者之间存在较好的线性关系 (y=7.4744x,R2=0.9989),以此为pH反馈补料工艺中补料液中葡萄糖与氨的混合比例,能较好地将谷氨酸发酵过程中葡萄糖浓度稳定在12~21 g/L。比较恒定葡萄糖浓度补料工艺与pH反馈补料工艺发现,采用pH反馈补料工艺进行发酵,葡萄糖转化率、谷氨酸产酸速率分别提高了9.06％和17.5％左右,同时发酵周期缩短2 h以上。     

基于遗传算法的谷氨酸发酵动力学参数估计

把遗传算法应用于求解谷氨酸分批发酵动力学模型参数,取交叉概率Pc=0．8、变异概率Pm=0．06、初始种群为20、遗传世代数为200代,能进一步提高谷氨酸分批发酵过程状态变量的计算值与实验值的吻合程度。模拟值与实验值对比显示,该动力学模型能很好地反映谷氨酸分批发酵过程。     

稀土元素对谷氨酸发酵的影响

通过对谷氨酸发酵培养基中添加稀土元素的研究,确定了几种能对发酵产酸有提高作用的稀土元素及其浓度,其中La^3 、Ce^4 、Nd^3 离子浓度分别为100mg/L、10mg/L、1mg／L时能提高产酸水平,而Sm^3 、Y^3 离子对发酵基本上没有影响。     

高分子量聚谷氨酸的微生物合成及其发酵过程的研究

目的：以枯草芽孢杆菌纳豆亚种为出发菌株,考察不同碳氮源及NaCl浓度、谷氨酸、种龄、接种量对微生物发酵产1-聚谷氨酸的影响,以提高γ-聚谷氨酸的产量。方法：该菌菌种活化后,接入种子培养基,于37℃、200r／min震荡培养18h,然后按2％接种量接入不同发酵培养基进行发酵培养。γ-聚谷氨酸分离纯化后,根据其产量筛选最适发酵培养基组成及发酵条件,并对产物进行分析测定。结果：①最佳碳氮源分别为葡萄糖、蛋白胨,NaCl浓度为30g／L、种龄15h、接种量3％,且需在培养基中添加谷氨酸。②该菌株在最适条件下发酵56h时,γ-聚谷氨酸产量达32．7g／L,凝胶渗透色谱分析其相对分子质量为426kDa,呈多分子质量聚集体形式。③γ-聚谷氨酸的合成与菌体生长并非完全同步。结论：γ-聚谷氨酸作为一种天然的、可生物降解的、对环境和人体无害的多聚物,可由微生物发酵合成,且在此适宜条件下产量较高。     

Effect of L-Aspartic Acid and L-Glutamic Acid on Production of L-Proline

To elucidate the effect of aspartic acid on growth of Kurthia catenaforma during the proline fermentation, this organism was compared with other bacteria with respect to the rate of consumption of aspartic acid, and to the activities of enzymes concerned in the metabolism of aspartic acid. Although no marked difference in enzyme activities was observed, the aspartic acid consumption rate of K. catenaforma was markedly higher than that of other organisms. The consumption of glutamic acid by K. catenaforma was not detected at 24 hr of culture. The difference between the consumption of aspartic acid and glutamic acid in this strain might result from a difference in permeability to the amino acids. We considered that L-glutamic acid might substitute for L-aspartic acid if the uptake of glutamic acid could be increased. A number of detergents were screened for their effect on consumption of glutamic acid. Cetyltrimethylammonium bromide, sodium laurylphosphate, and polyoxyethylene sorbitan monolaurate were found to increase the transport rate of glutamic acid, but not of aspartic acid. A method of producing L-proline from glutamic acid was established with the aid of detergents.     

富锌产γ-氨基丁酸乳酸菌的筛选及初步鉴定

用含0.2%-0.25%Zn^2＋的MRS培养基从发酵酸菜、酸奶中分离出6株具富锌能力细菌,经形态和生理生化特性鉴定,初步判断菌株为乳酸菌。采用纸层析检测MRSG培养基培养的6株菌上清液后,初步确定有4株能产γ-氨基丁酸。以此4菌株基因组为模板PCR扩增到540 bp的片段,经测序和序列分析,证明是谷氨酸脱羧酶基因高度保守区域,初步证明谷氨酸脱羧酶基因的存在。此4株菌发酵液经HPLC检测和分析后,证实2号菌株能转化谷氨酸钠生成GABA,产量为4.8 g/L,在应用方面具有很大的潜力。     

Glutamic Acid Production in Biotin-rich Media by Temperature-sensitive Mutants of Brevibacterium lactofermentum,a Novel Fermentation Process

Temperature-sensitive mutants were derived from Brevibacterium lactofermentum strain 2256 in a search for mutants which would produce a large amount of L-glutamic acid in biotin- rich media at the nonpermissive temperature. A total of 159 mutant strains was selected which showed adequate growth at 30°C but showed little or no growth at 37°C on minimal medium. Twenty of these were found to produce glutamic acid in a biotin-rich medium after a temperature shift from 30°C to 37°C, while the wild-type strain 2256 did not produce it under the same cultural condition.

One of the typical mutant strains, Ts-88, produced approximately 2g/dl of glutamic acid from beet molasses (the yield > 55%) in the presence of 33 µg/liter of biotin when tempera- , ture was shifted from 30°C to 40°C during the cultivation. It was concluded that, by controlling only temperature during fermentation, glutamic acid production could be realized in media containing biotin-rich natural carbon sources, without any chemical control such as the addition of expensive surface-active agents or antibiotics. Characteristics and merits of the novel fermentation process are discussed.     

Mechanism of Proline Production by Kurthia catenaforma

The fate of aspartic acid used for proline fermentation by Kurthia catenaforma was traced by using aspartic acid-U-(14)C. The radioactivities of proline and glutamic acid increased with the disappearance of aspartic acid. After 40 hr, aspartic acid disappeared from the medium and radioactive alpha-ketoglutaric acid was detected. The radioactivity of proline reached 44% of aspartic acid radioactivity at 40 hr. The specific radioactivities of these amino acids and of alpha-ketoglutaric acid supported the notion that proline is produced mainly from aspartic acid via alpha-ketoglutaric acid and glutamic acid. Since the levels of glutamic acid dehydrogenases (EC 1.4.1.2 and EC 1.4.1.4) were low in this organism, it appears that the nitrogen atom of aspartic acid enters proline by the action of aspartate aminotransferase (EC 2.6.1.1). The mechanism of proline production is discussed on the basis of the role of aspartic acid in this fermentation.     

Kinetics of molecular weight reduction of poly(glutamic acid) by in situ depolymerization in cell-free broth of Bacillus subtilis

Poly(glutamic acid) (PGA) is a water soluble, biodegradable biopolymer that is produced by microbial fermentation. Recent research has shown that poly(glutamic acid) can be used in drug delivery applications for the controlled release of paclitaxel (Taxol) in cancer treatment. The molecular weight of microbial poly(glutamic acid) is generally larger than what is required for drug delivery. As such, molecular weight reduction is a necessary step in producing poly(glutamic acid) for this application. Poly(glutamic acid) produced by Bacillus subtilis IFO 3335 was subjected to in situ depolymerization in the cell-free fermentation broth. Molecular weight reduction was measured, and an empirical kinetic model was used to correlate the experimental data. The kinetic rate constant, k, was found to be 6.92 × 10⁻⁶ h⁻¹ at pH 7.0 and 37 °C, which were the optimum depolymerization conditions.     

Glutamic acid and by-product synthesis by immobilized cells of the bacterium Corynebacterium glutamicum

Summary The time course for the synthesis of glutamic acid and by-products from glucose was investigated using immobilized cell reactor of the bacterium C.glutamicum. Lactic acid, succinic acid, alanine acid and aspartic acid were formed early in the fermentation and during the active growth phase, whereas gluconic acid, -ketoglutaric acid and proline were produced late and during the active phase of glutamic acid synthesis. Oxygen transfer rate in fermentation broth had a pronounced effect on the nature and quantities of fermentation products. In continuous fermentation and at OTR of 102.5 mMO₂/l.h., formation of by-products greatly decreased and up to 58.5 g/l of glutamic acid were produced with a conversion efficiency of 74.6% of the theoretical value and volumetric productivity of 6.2 g/l.h.