在线推定和控制葡萄糖浓度改善谷氨酸发酵性能

谷氨酸发酵过程一般需要定时、人工分批式地添加葡萄糖。该流加操作方式会引起发酵罐内葡萄糖浓度的剧烈波动, 不利于高效、稳定的谷氨酸生产。谷氨酸发酵具有显著的非增殖耦联特征, 产酸期葡萄糖耗量与氨水耗量存在非常明显的关联性。通过在线计量氨水耗量推定糖耗以及葡萄糖浓度, 可比较准确地将谷氨酸发酵产酸期的糖浓度控制在预先设定的水平。当糖浓度控制在5 g/L~10 g/L的低水平时, 最终谷氨酸浓度可以达到80 g/L的较高水平, 高糖浓度下的渗透压效应有望得到缓解, 有利于发酵生产的稳定。     

酶电极流动注射分析法测定葡萄糖

 一种酶电极流动注射分析系统(EFIA)用于血糖和发酵葡萄糖的快速测定。研究了酶电极及其工作系统的性能和各种影响参数,,奠定了实用化基础。     

用葡萄糖酶电极法测定葡萄糖淀粉酶活性的研究

利用固定化葡萄糖氧化酶酶膜和过氧化氢电极组成酶电极测定葡萄糖淀粉酶的活性单位。用已知单位的葡萄淀粉酶作为测定标准定标后,在仪器上直接测出被测样品的葡萄糖淀粉酶活性单位,测定时间１４０ｓ,操作周期３ｍｉｎ,连续１０次测定ＣＶ值为０.６７％,５０～５００ｕ／ｍｌ的范围内线性良好,ｒ＝０.９９９９。     

谷氨酸发酵的在线控制

谷氨酸是我国大宗的发酵产品之一.谷氨酸发酵需要补糖,传统的方法是凭经验,瞬时、间歇式地投料,但这种间歇式补料容易引起糖浓的剧烈变化,造成罐内环境、如渗透压等的不稳定,从而对菌体代谢活性及最终发酵生产带来负面影响~([1-3]).     

葡萄糖测定方法的比较研究

比较了传统斐林定糖,葡萄糖氧化酶－过氧化物酶比色法,葡萄糖氧化酶电极自动分析仪法测定葡萄糖。比较测定了的结果显示,三法的平均标准误差（ＳＤ）,变异系数（ＣＶ）均十分接近。通过对此三种方法的回归相关性分析显示：斐林法－酶终点比色法的回归方程为ｙ＝０．９８４３ｘ＋６．３２３９,相关系数Ｒ＾２＝０．９９８９,斐林法－自动分析仪法的回归方程为ｙ＝１．００８８ｘ＋２．０４８３,相关系数Ｒ＾２＝０．９９９１,     

一种免受乙醇干扰的GOD的酶电极

将酶电极应用于发酵糖的测定时。与糖共存的乙醇常常会影响测糖的准确性,通过对葡萄糖氧化酶(GOD)电极的研究,探讨了乙醇对测糖酶电极测定影响,进而研制出抗干扰的GOD酶电极,若是GOD电极的酶膜通过夹心法制备．在乙醇含量为0．1％(V／V)时·即产生显著的影响,使测定结果偏大4．3%,且乙醇的影响随浓度的升高而增大,若用尼龙网固定GOD膜,GOD电极在测定20mmol／L和5mm01／L左右的葡萄糖溶液时,含量高达9％的乙醇仍未对测定产生显著的影响．表现出良好的不受乙醇干扰的特性,并且,该尼龙网GOD电极具有良好的重复性、稳定的响应活性及较长的保存期．     

尿素氮-葡萄糖双功能分析仪的研究

由固定化脲酶、谷氨酸脱氢酶、谷氨酸氧化酶、葡萄糖氧化酶的复合酶膜组成的双电极系统,可以同时测定尿素氮和葡萄糖的含量,每次进样量为25μl,20s即可测定出尿素氮和葡萄糖的含量。在0～60mg/dl尿素氮、0～500mg/dl葡萄糖范围内具有良好的线性关系。连续测定20次的变异系数分别为1.02％和1.05％。酶膜使用寿命为两星期以上。此仪器可广泛应用于临床检验和体育训练中。     

酶电极法快速测定甘油含量的研究

利用酶固定化技术,以甘油激酶（GK）、甘油-3-磷酸氧化酶（GPO）为反应酶,研究GK、GPO的固定化方法及固定化模式,制备甘油酶膜、甘油酶电极,并利用其测定甘油含量。结果表明,GK、GPO按1：1比例固定化时,酶电极电流信号最高;最高效固定模式为：GK固定于核微孔膜,共价偶联GPO固定于Biodyne膜,形成共价双酶膜,进而组装为甘油酶电极。性能研究表明,甘油酶电极最适pH值为7.0,最佳温度为28~32℃;最佳实验条件下,线性范围为0.05~9.00 g/L;回收率为98.4%~102.4%,稳定性高,相对标准偏差（RSD）<5%;测定结果与高效液相色谱法、高碘酸氧化法比较,无明显差异（P>0.05）,且该方法操作简单,专一性强,检测快速,适于实际生产中甘油的实时定量及监控。     

重组大肠杆菌高密度培养研究进展

重组大肠杆菌细胞高密度培养(High cell-density cultivation,HCDC)是获得高外源蛋白产率的一种重要策略,影响重组大肠杆菌高密度培养的因素主要有以下几个方面:重组体的构建及其稳定性,培养基成分,培养方式,培养条件及培养过程中抑制性代谢产物的积累等。从以上几个方面对近期的研究进展进行了综述。     

不同流加发酵方式对重组大肠杆菌细胞外Hegf表达水平的影响

人表皮生长因子 (ｈＥＧＦ)是由 5 3个氨基酸组成的单链多肽 ,内有 3个二硫键。它具有多种重要的生物学功能 ,能够促进表皮细胞的生长繁殖 ,加速皮肤和角膜创伤的修复 ,加速胃溃疡的治疗和抑制胃酸的分泌 ,具有广阔的医疗应用前景[1 ] 。最近又发现ｈＥＧＦ在化妆品产业中有潜在的重大应用[2 ] 。这些都促进了人们试图采用基因工程技术大规模地生产ｈＥＧＦ。1982年Ｓｍｉｔｈ等人首先在Ｅ .ｃｏｌｉ中以融合蛋白形式实现了ｈＥＧＦ的表达[3 ] ,随后在枯草杆菌和酵母菌中亦相继实现了表达[4] 。由于ｈＥＧＦ在酵母菌中表达水平很低 ,大肠杆…     

Mineralization of pentachlorophenol in a contaminated soil by Pseudomonas sp UG30 cells encapsulated in κ-carrageenan

A fermentation process in Escherichia coli for production of supercoiled plasmid DNA for use as a DNA vaccine was developed using an automated feed-back control nutrient feeding strategy based on dissolved oxygen (DO) and pH. The process was further automated through a computer-aided data processing system to regulate the cell growth rate by controlling interactively both the nutrient feed rate and agitation speed based on DO. The process increased the total yield of the plasmid DNA by approximately 10-fold as compared to a manual fed-batch culture. The final cell yield from the automated process reached 60 g L⁻¹ of dry cell weight (OD₆₀₀ = 120) within 24 h. A plasmid DNA yield of 100 mg L⁻¹ (1.7 mg g⁻¹ cell weight) was achieved by using an alkaline cell lysis method. Plasmid yield was confirmed using High Performance Liquid Chromatography (HPLC) analysis. Because cells had been grown under carbon-limiting conditions in the automated process, acetic acid production was minimal (below 0.01 g L⁻¹) throughout the fed-batch stage. In contrast, in the manual process, an acid accumulation rate as high as 0.36 g L⁻¹ was observed, presumably due to the high nutrient feed rates used to maintain a maximum growth rate. The manual fed-batch process produced a low cell density averaging 10–12 g L⁻¹ (OD₆₀₀ = 25–30) and plasmid yields of 5–8 mg L⁻¹ (approximately 0.7 mg g⁻¹ cells). The improved plasmid DNA yields in the DO- and pH-based feed-back controlled process were assumed to be a result of a combination of increased cell density, reduced growth rate (μ) from 0.69 h⁻¹ to 0.13 h⁻¹ and the carbon/nitrogen limitation in the fed-batch stage. The DO- and pH-based feed-back control, fed-batch process has proven itself to be advantageous in regulating cell growth rate to achieve both high cell density and plasmid yield without having to use pure oxygen. The process was reproducible in triplicate fermentations at both 7-L and 80-L scales. Received 22 March 1996/ Accepted in revised form 20 September 1996     

Model based calculation of substrate/inducer feed-rate profiles in fed-batch processes for recombinant protein production

A model-based feed-rate profile optimization problem is discussed for the fed-batch recombinant protein production. Two optimization procedures, an evolutionary programming technique and a simplified method using the dynamic programming concept, are discussed and compared. Modeling as well as experimental results are presented.     

过量表达自身hemA基因的重组大肠杆菌发酵生产5-氨基乙酰丙酸的研究

研究了优化重组大肠杆菌产5-氨基乙酰丙酸（ALA）的条件,提高大肠杆菌发酵生产AL气的产量。在测定重组大肠杆菌GT48的生长曲线的基础上,确定诱导时间,优化摇瓶发酵条件。然后,进一步在5L发酵罐上进行间歇和流加发酵研究。摇瓶实验表明,细胞培养最佳初始pH为6．5,最佳诱导时间为稳定期前期,最佳接种量为2％,过高的葡萄糖浓度对细胞生长和产物合成均有一定的抑制作用。在5L发酵罐间歇发酵中,重组菌产ALA能力达到47．8mg／L。采用流加发酵可以进一步将产物产量提高到63．8mg／L。构建的过量表达自身的hemA基因的大肠杆菌具有较高的产ALA能力,通过发酵条件优化和采用流加发酵可以提高AL气产量。     

Glucose-containing polymer rings enable fed-batch operation in microtiter plates with parallel online measurement of scattered light,fluorescence, dissolved oxygen tension,and pH

Bioprocesses operated in batch mode can induce adverse effects like overflow metabolism, substrate inhibition, osmotic inhibition, oxygen limitation, and catabolite repression. To avoid these adverse effects, fed-batch is the predominant operation mode in industrial production. Nevertheless, screening for optimal production strains is usually performed in microtiter plates and shake flasks operated in batch mode without any online monitoring. Recently, a polymer-based controlled-release fed-batch microtiter plate with stable glucose release characteristics was described. In this study, a glucose-containing polymer matrix was used to manufacture polymer rings that were placed at the bottom of a 48-well microtiter plate. Thereby, the liquid content of the well became accessible for optical measurement by the BioLector device. Reflections caused by the polymer ring were minimized by adjusting the scattered-light measurement position. Influences on the measurement of the dissolved oxygen tension and pH could be avoided by choosing appropriate polymer-ring geometries. These adjustments enabled parallel online measurement of scattered light, fluorescence, dissolved oxygen tension, and pH of Escherichia coli BL21 (DE3) fed-batch cultivations. The online monitoring and fed-batch operation capabilities of the fed-batch microtiter plate presented in this study finds optimal application in screenings and initial process development.