谷氨酸发酵的智能控制

本文在控制策略上,对谷氨酸发酵过程的温度和pH值控制系统分别采用了非线性、变比例增益的实际微分PID算法与智能——模糊控制算法。现场投运表明,此微机控制策略正确、有效,控制软件智能较高,能缩短发酵周期,提高产酸2.26％,节省尿素5％。该成果获得90年广东省高教三等奖。     

L-谷氨酸发酵的变温控制工艺研究

以黄色短杆菌GDK-9为供试菌株,首先考察了不同的培养温度(30~36℃)对L-谷氨酸摇瓶发酵的影响。结果发现,在34℃条件下菌体生长情况最好,且L-谷氨酸产量最高,可达86g/L。基于上述结果,研究了3种变温控制模式下L-谷氨酸摇瓶发酵过程。得出如下结论:0~6h发酵温度为34℃,6h后每隔6h提高1℃,并采用这种温度控制方式进行5L自控发酵罐试验,36h后可积累L-谷氨酸130g/L,比同类研究的产量(102 g/L)提高了27%。     

表面活性剂作为谷氨酸发酵促进剂的研究

通过对预先选择的7种表面活性剂作为谷氨酸发酵促进剂效果的初步筛选,得知两性甜菜碱449和芹菜素硬脂酸酯对产酸率提高有较为明显的效果;进而从加入时间、加入剂量两个方面考察了筛选出的两种表面活性剂对谷氨酸棒状杆菌(Corynebacterium glutamicum)T-613菌体生长和谷氨酸发酵产酸的影响,考察了两种表面活性剂同时加入的协同效果,得到了最佳发酵工艺参数。研究结果表明,菌种对数生长期(第3 h)加入质量分数为10%的两性甜菜碱449型表面活性剂2.5 mL,发酵中后期(第12 h)加入质量分数为0.5%的芹菜素硬脂酸酯0.8 mL,在不影响菌种生长的前提下,促使谷氨酸产酸率由3.05%提高到3.76%,较原产酸率提高了23.28%,发酵周期由44 h下降为36 h,缩短了8 h。     

谷氨酸发酵过程先进控制

阐述基于工业过程计算机和PLC的谷氨酸发酵过程控制,在若干生化过程理论模型的实际应用具有一定困难的情况下,研究了在发酵过程中对某些生化过程变量的估算方法及相应的先进过程控制策略,对谷氨酸发酵过程的主要控制回路作了介绍。通过对50m3发酵罐实时控制,表明本文所述控制系统的有效性。     

大型谷氨酸发酵过程的DCS控制

介绍了μＸＬ型集散控制系统在大型谷氨酸发酵罐实时控制方面的应用，对稳定发酵环境参数、提高产酸率收到了良好的效果。对某些生化参数的推断做了初步尝试。     

谷氨酸发酵废液提取核糖核酸的开发研究

在味精的生产过程中,占谷氨酸发酵分离后的废液1～2％的菌泥可用来提取RNA。本文叙述了稀碱法、苯酚法和浓盐法提取RNA的工艺,并简介了RNA在医疗、制药、农业和食品方面的用途。这是一项有前途的对污水治理的有效方法,也是一项对成本低的新资源新产品的开发。     

对正交设计法安排小磨试验的一点看法

正交试验是利用正交表进行试验设计,通过少量试验获得较多信息,找出最好或满意的试验条件的常用方法,水泥企业为了得出本厂的混合材、石膏最佳掺量,需按一定的配比方案来安排小磨试验时常采用此法。历版的《水泥企业化验室工作手册》(下简称《手册》)中对此方法都有专门的介绍[     

艰难梭状芽孢杆菌谷氨酸脱氢酶的原核表达、纯化及其酶活性

目的原核表达艰难梭状芽孢杆菌(Clostridium difficile,CD)谷氨酸脱氢酶(Glutamate dehydrogenase,GDH),纯化重组蛋白,并检测其酶活性。方法采用PCR法从标准株ATCC BAA-1805中扩增GDH基因,克隆至原核表达载体pET-28a(+)中,构建重组表达质粒pET-28-GDH,转化感受态大肠杆菌Rosetta(DE3),IPTG诱导表达。表达的重组蛋白经SDS-PAGE和Western blot分析,并通过Ni-NAT层析柱分离纯化后,根据脱氢酶酶活测定方法,测定纯化蛋白的酶活性。结果重组表达质粒pET-28-GDH经双酶切及测序,证实构建正确,重组蛋白的相对分子质量约46 000,可与抗His单抗特异性结合,主要以可溶性形式表达。纯化的目的蛋白纯度可达90%,浓度为1.7 mg/ml。纯化蛋白GDH以NADPH和NADP为辅酶,酶活性分别为42.6和63.3 U/L。结论已成功在大肠杆菌中表达了重组CD GDH,纯化的目的蛋白纯度高,具有GDH酶活性,为制备GDH单克隆抗体奠定了基础。     

正交试验设计法

在生产和科学试验中,为了改革工艺和研制新产品,通常要做很多多因素试验。因此在试验之前必须好好“设计”一番。以最大限度地减少实验次数,缩短试验周期,同时又能获得明确可靠的结论。这里介绍一种正交试验设计法。     

枯草芽孢杆菌HG-02聚谷氨酸发酵条件优化

筛选得到了一株高产γ-聚谷氨酸的枯草芽孢杆菌HG-02。通过单因素试验和正交实验对发酵培养基和培养条件进行了优化,小试发酵验证结果显示,优化后γ-聚谷氨酸的发酵质量浓度达35.9 g/L。优化后的发酵培养基为：蔗糖4%,蛋白胨3.5%,谷氨酸钠6%,硫酸镁0.1%,氯化钠0.5%,氯化钙0.01%,磷酸氢二钾0.05%,硫酸锰0.02%,消泡剂1%。优化后的培养条件为：初始pH7.0,装液量40%,接种量6%,摇床转速为220 r/min, 37℃恒温培养48 h。     

味精闭环生产新工艺

介绍通过膜分离技术开发出来的味精闭环生产工艺路线。该工艺采用无机陶瓷超滤膜将发酵液超滤;用浓硫酸水解截留液;过滤水解液,在滤液中得水解产物;混合渗透液与水解过滤液,进行谷氨酸结晶;最后用味精结晶母液纯化谷氨酸结晶。该工艺采用蒸发与冷却耦合连续结晶工艺,结晶收率高,且节能效果显著。     

抗生素AGPM摇瓶发酵条件的正交实验

采用正交实验设计考察了培养基组成对新型抗生素AGPM发酵的影响,改进后在培养基组成为(g/L)：葡萄糖5, 玉米淀粉40, 黄豆饼粉16, 玉米浆2 ml, K2HPO4 1.0, MgSO4×7H2O 0.5, NaCl 0.5, 淀粉酶0.05及pH 5.5的条件下,单位发酵液的活性提高了18.9倍;同时表明较高的碳氮比对抗生素AGPM的合成有利.     

过量表达琥珀酸输出蛋白SucE对谷氨酸棒杆菌厌氧生产琥珀酸的影响

谷氨酸棒杆菌Corynebacterium glutamicum ATCC 13032在厌氧条件下通过三羧酸循环还原臂合成琥珀酸。高效地将细胞内的琥珀酸输出到胞外,对琥珀酸的生物合成具有重要意义。本实验以C.glutamicum XZ为出发菌株过表达琥珀酸输出蛋白表达基因suc E。突变株C.glutamicum XZ(p Exhsuc E)的琥珀酸得率提高了7%,生产率提高了19%。通量分析表明,过表达suc E基因后乙醛酸循环的相对通量提高了50%。采用两阶段培养,突变株C.glutamicum XZ(p Exhsuc E)的琥珀酸产量达到518 mmol·L-1,厌氧阶段的比生产率为0.95 mmol g CDW-1·h-1,1 mol glucose的平均得率为1.5 mol。     

正交实验设计优化穿心莲总黄酮提取工艺

探讨了用乙醇回流提取穿心莲黄酮类化合物,找出最佳提取条件。通过正交实验探索乙醇回流法提取穿心莲黄酮类化合物的最佳工艺条件,采用紫外分光光度法测定总黄酮含量。影响实验因素的主次顺序是：固液比〉浸提时间〉乙醇浓度〉浸提温度,其中固液比对实验指标有显著性的影响。最佳提取工艺为固液比1：20、浸提时间4h、乙醇浓度60％、浸提温度50℃。     

基因敲除谷氨酸棒杆菌提高鸟氨酸产量及比较蛋白质组学分析(英文)

Engineered Corynebacterium glutamicum was constructed for L-ornithine production by disrupting genes of argF and proB to prevent the flux away from L-ornithine.Effect of the inactivation of 2-oxoglutarate de-hydrogenase complex(ODHC) on L-ornithine production was also investigated.It was found that the inactivation of ODHC by knockout of the kgd gene enhanced L-ornithine production.The engineered C.glutamicum ATCC13032(ΔargFΔproBΔkgd) produced L-ornithine up to 4.78 g·L-1 from 0.24 g·L-1 of the wild-type strain.In order to understand the mechanism of L-ornithine production in C.glutamicum ATCC13032(ΔargFΔproBΔkgd) and find out new strategies for further enhancing L-ornithine production,the comparative proteome between the wild-type and the engineered strain was analyzed.L-Ornithine overproduction in the engineered strain was related to the up-regulation of the expression levels of enzymes involved in L-ornithine biosynthesis pathway and down-regulation of the expression levels of proteins involved in pentose phosphate pathway.The overexpression of genes in the upstream pathway of glutamate to increase the availability of endogenous glutamate may further in-crease ornithine production in the engineered C.glutamicum and the ornithine synthesis enzymes(ArgCJBD) may not be the limiting enzymes in the engineered C.glutamicum.     

正交试验法优选黄芪多糖的水提工艺研究

以浸膏得率和多糖含量为检测指标,利用L9(34)正交试验法对黄芪的水提工艺进行了优选。确定最佳的水提条件为:先用10倍量的水浸泡0.5h,煎煮1.5h;第2次加8倍量水,煎煮1.0h;第3、4次各加6倍量水,煎煮时间分别为1.0h。本法快速,简便,准确,适用于黄芪药材的质量控制。     

Fermentative Production of Halogenated Tryptophan Derivatives with Corynebacterium glutamicum Overexpressing Tryptophanase or Decarboxylase Genes

The aromatic amino acid l -tryptophan serves as a precursor for many valuable compounds such as neuromodulators, indoleamines and indole alkaloids. In this work, tryptophan biosynthesis was extended by halogenation followed by decarboxylation to the respective tryptamines or cleavage to the respective indoles. Either the tryptophanase genes tnaAs from E. coli and Proteus vulgaris or the aromatic amino acid decarboxylase genes AADCs from Bacillus atrophaeus, Clostridium sporogenes, and Ruminococcus gnavus were expressed in Corynebacterium glutamicum strains producing (halogenated) tryptophan. Regarding indoles, final titers of 16 mg L⁻¹ 7-Cl-indole and 23 mg L⁻¹ 7-Br-indole were attained. Tryptamine production led to a much higher titer of 2.26 g L⁻¹ upon expression of AADC from B. atrophaeus. AADC enzymes were shown to be active with halogenated tryptophan in vitro and in vivo and supported production of 0.36 g L⁻¹ 7-Br-tryptamine with a volumetric productivity of 8.3 mg L⁻¹ h⁻¹ in a fed-batch fermentation.     

Expert System for the Design of an Industrial Fermentation Plant for the Production of Alcohol

A user-friendly expert system was developed for the design of large scale continuous fermentation units for alcohol production. This expert system utilized an intensive rule-based program, with a knowledge base structured using both expert knowledge data and industrial practices, as well as computer simulations. The simulation algorithm was developed from mass and energy conservation of continuous multi-staged plants and kinetic equations representing the fermentation process by Saccharomyces cerevisiae. The kinetic parameters were adapted from industrial data, leading to a reliable mathematical model that represented the industrial process reasonably well. Since the system was designed for continuous processes with several stirred tanks connected in series, the optimal volume profile and number of stages were previously optimized. The system developed could be utilized either for the design of industrial plants or for the improvement and modernization of existing plants. This system was shown to be reliable since the difference between theoretical calculations and practical results was minimal. © 1997 SCI.     

Reconstruction of the Diaminopimelic Acid Pathway to Promote L-lysine Production in Corynebacterium glutamicum

The dehydrogenase pathway and the succinylase pathway are involved in the synthesis of L-lysine in Corynebacterium glutamicum. Despite the low contribution rate to L-lysine production, the dehydrogenase pathway is favorable for its simple steps and potential to increase the production of L-lysine. The effect of ammonium (NH₄⁺) concentration on L-lysine biosynthesis was investigated, and the results indicated that the biosynthesis of L-lysine can be promoted in a high NH₄⁺ environment. In order to reduce the requirement of NH₄⁺, the nitrogen source regulatory protein AmtR was knocked out, resulting in an 8.5% increase in L-lysine production (i.e., 52.3 ± 4.31 g/L). Subsequently, the dehydrogenase pathway was upregulated by blocking or weakening the tetrahydrodipicolinate succinylase (DapD)-coding gene dapD and overexpressing the ddh gene to further enhance L-lysine biosynthesis. The final strain XQ-5-W4 could produce 189 ± 8.7 g/L L-lysine with the maximum specific rate (q_Lys,max.) of 0.35 ± 0.05 g/(g·h) in a 5-L jar fermenter. The L-lysine titer and q_Lys,max achieved in this study is about 25.2% and 59.1% higher than that of the original strain without enhancement of dehydrogenase pathway, respectively. The results indicated that the dehydrogenase pathway could serve as a breakthrough point to reconstruct the diaminopimelic acid (DAP) pathway and promote L-lysine production.     

南极假丝酵母产脂肪酶在15 L发酵罐中培养条件的研究

对15 L发酵罐中南极假丝酵母(C.antarctica DSM3855)的发酵工艺条件进行了研究,确定了最适的产酶工艺条件:搅拌速度为300 r·min-1,通气量为10 L·min-1,接种量为10%,添加GPE作为消泡剂.在此培养条件下,培养54 h产酶达到高峰值,最高酶活力达到19.3 U·mL-1.