L-色氨酸中试生产研究

为提高L-色氨酸发酵水平,在100L发酵罐上考察了不同操作条件对大肠杆菌发酵生产L-色氨酸的影响。结果表明不同操作条件对L-色氨酸发酵有显著影响。残糖浓度、补料方式、柠檬酸钠添加质量分数、接种量及溶氧量分别控制在5 g/L、溶氧控制脉冲补料、2 g/L、5%及10%～15%时,发酵效果最好,在1 m3罐上进行验证实验,L-色氨酸产量稳定在40 g/L左右。     

中后期不同溶氧水平对赖氨酸发酵的影响

发酵过程中合理且充分供氧是提高L-赖氨酸发酵产量的主要途径之一,实际生产中溶氧控制策略又往往难以进行具体化操作,尤其是发酵中后期的溶氧水平直接决定了发酵生产的水平.通过5 L发酵罐实验分析中后期溶氧水平对赖氨酸发酵的影响,研究了不同溶氧水平条件下的发酵规律.实验数据表明,赖氨酸发酵中后期溶氧水平控制在60%~80%内,可以明显提高发酵的终点酸、转化率、酸固比和单罐总酸等生产指标,与正常控制水平(40%~60%)相比,依次分别提高了7.30%、3.08%、0.92%、5.48%.     

用于谷胱甘肽合成的重组大肠杆菌的培养和诱导表达

研究了用于谷胱甘肽合成的温度诱导型重组大肠杆菌的补料分批发酵,通过葡萄糖的脉冲补入,结合溶氧反馈控制技术,实时控制发酵过程中乙酸的产生. 在此基础上,进一步考察了在谷胱甘肽合成酶系诱导表达阶段,流加酵母粉和蛋白胨时重组大肠杆菌的补料分批发酵过程. 结果发现,脉冲补料-溶氧反馈控制技术在流加酵母粉和蛋白胨的补料分批发酵中仍能得到很好的应用,乙酸浓度可被有效地控制在2.5 g/L以下. 通过比较诱导表达阶段流加不同组成的补料液成分,发现添加适量酵母粉和蛋白胨可促进谷胱甘肽合成酶系的表达,在42℃诱导4 h收获的重组大肠杆菌酶法合成谷胱甘肽达到2.46 g/L.     

黑曲霉葡萄糖酸发酵的工艺控制方法

针对葡萄糖酸生产菌--黑曲霉,在10 L发酵罐进行了其发酵工艺控制研究.结果表明,高含量葡萄糖对黑曲霉生成葡萄糖酸没有抑制作用,较好的葡萄糖控制方式为维持葡萄糖含量20%左右,总糖含量为30%.流加30%NaOH溶液、流加25%～30% Ca(OH)2乳浊液、分批添加CaCO3等3种方式都能很好地控制pH值,但各有适用范围;通过调整搅拌转速的方式维持溶氧含量50%左右,可提高产葡萄糖酸性能,并能降低搅拌能耗.     

黑曲霉发酵生产壳聚糖工艺研究

采用生物发酵法制备了壳聚糖.在常用培养基中添加酒糟进行了正交实验,确定培养基组成为:玉米浆4%、烘干磨碎酒糟4%、葡萄糖3%、硫酸镁1%.得到了摇瓶发酵和15 L发酵罐中的发酵工艺.对发酵罐发酵过程的特征参数进行了初步研究,分析了生物量、pH值、残糖含量、Mg2 含量的变化,并绘制出变化曲线.     

间歇流加L-山梨糖发酵生产2-酮基-L-古龙酸研究

利用氧化葡萄糖酸杆菌(Gluconobacre roxydans)和蜡状芽孢杆菌(Bacillus cereus)混合菌系及自动控制温度、pH和溶氧系统,在发酵过程中间歇流加L-山梨糖发酵生成2-酮基-L-古龙酸(2-KLG).结果表明:间歇流加L-山梨糖可以解除高浓度糖对产酸的抑制作用,提高了糖的转化率,当将L-山梨糖的终浓度调高到14%(w/V)时,2-KLG产量为130 mg/mL左右,转化率达90%,发酵周期40～60 h之间,发酵周期略有延长.     

维生素C发酵一步工艺优化研究

针对目前维生素C（VC）发酵过程中一步发酵生产进行工艺优化探讨,从pH值、种后溶氧、山梨醇投醇浓度等关键生长条件入手,达到缩短发酵周期,提高产糖速率的效果。结果表明：一步菌种发酵过程最适pH值为4.8～6.0,发酵周期与种后溶氧成线性关系,投醇浓度控制在30%以下产糖速率最快。     

动胶菌发酵生产可降解塑料PHAs

选育能高产聚羟基烷酸的优良菌株,采用最佳的发酵条件是实现细菌发酵法生产可降解塑料工业化应用的关键因素之一。因此对分离自活性污泥中的菌株Zoogloeasp.GY3进行了3L发酵罐的发酵条件研究。结果表明,罐的通气量、转速是影响最终发酵结果的主要因素,其最佳条件为在装液量为1800mL的条件下,通气量为1L/(L.min),20h之前的最佳搅拌速度为350r/min,20h之后的最佳搅拌速度为500r/min。流加控制发酵液糖浓度维持在2.5%,发酵结束后细胞生物量高达53.74g/L,PHAs质量浓度为45.9g/L,PHAs含量提高到85.4%。     

30L发酵罐培养枯草芽孢杆菌产高密度芽孢的研究

通过本实验室优化的培养基配方,结合前期流加葡萄糖液,使残糖维持在0.5%以下,溶氧和转速关联的培养方式,在30L的发酵罐中枯草芽孢杆菌菌数和芽孢较快达到9次方,培养26h,菌数最高能达到1.2×1010个/m L,芽孢最高能达到1.1×1010个/m L。     

VC二步发酵高糖流加工艺的研究

利用从生产中选育得到的耐高糖菌株,通过在线的检测数据,应用改进的高浓度山梨糖流加控制工艺,考察了初始山梨糖浓度、底物浓度、流加阶段pH等对维生素C二步发酵的影响,确定了适宜的控制工艺,提高了发酵水平及设备利用率.     

产朊假丝酵母流加发酵法生产谷胱甘肽

研究了产朊假丝酵母在不同葡萄糖浓度下的谷胱甘肽(GSH)分批发酵过程,在此基础上进一步考察了重复补料、恒速流加和指数流加等不同培养方式对GSH发酵生产的影响. 结果发现,这几种培养方式都可以实现酵母细胞和GSH的高产. 综合比较,无论是从细胞还是从GSH的产量、得率和生产强度的角度来看,指数流加都是较为理想的选择. 经过48 h的指数流加培养,细胞干重达到40.9 g/L, GSH产量和胞内GSH含量分别达到857.2 mg/L和2.25%.     

Three-stage fermentation and kinetic modeling of bioflocculant by Corynebacterium glutamicum

Fermentation of bioflocculant with Corynebacterium glutamicum was studied by way of kinetic modeling. Lorentzian modified Logistic model, time-corrected Luedeking–Piret and Luedeking–Piret type models ...     

三段式谷氨酸棒杆菌发酵生产生物絮凝剂的过程及动力学模拟(英文)

Fermentation of bioflocculant with Corynebacterium glutamicum was studied by way of kinetic modeling. Lorentzian modified Logistic model, time-corrected Luedeking–Piret and Luedeking–Piret type models were pro-posed and applied to describe the cell growth, bioflocculant synthesis and consumption of substrates, with the correlation of initial biomass concentration and initial glucose concentration, respectively. The results showed that these models could well characterize the batch culture process of C. glutamicum at various initial glucose con-centrations from 10.0 to 17.5 g·L?1. The initial biomass concentration could shorten the lag time of cel growth, while the maximum biomass concentration was achieved only at the optimal initial glucose concentration of 16.22 g·L?1. A novel three-stage fed-batch strategy for bioflocculant production was developed based on the model prediction, in which the lag phase, quick biomass growth and bioflocculant production stages were sequentially proceeded with the adjustment of glucose concentration and dissolved oxygen. Biomass of 2.23 g·L?1 was obtained and bioflocculant concentration was enhanced to 176.32 mg·L?1, 18.62% and 403.63%higher than those in the batch process, respectively, indicating an efficient fed-batch culture strategy for bioflocculant production.     

分泌型重组人生长激素工程菌的高密度发酵

目的　研究葡萄糖剂补加策略 ,以获得大肠杆菌的高密度发酵和人生长激素 (HGH)的高表达。方法　采用批式发酵和补料批式发酵方式 ,葡萄糖的补加采用逐渐增加的方式 ,包括pH、DO和浓度控制。结果　获得HGH的表达量为 30 0mg/L ,光密度达到 12 4(A60 0 )。结论　在大肠杆菌中表达异源蛋白时 ,葡萄糖的补加方式是关键因素 ,补料批式发酵要优于批式发酵     

Analysis of fed-batch fermentation processes by graphical methods

Graphical methods are used for many types of engineering calculations. Their appeal is often related to calculational simplicity, but probably the increased understanding that accompanies a graphical calculation method is chiefly responsible for their popularity. In the present work existing graphical concepts which have been used in chemical reaction engineering and fermentation technology have been extended to allow the calculation of biomass and product formation for a variety of fermentation processes. Comparisons in productivity are made among chemostat, batch, fed batch with variable and constant feed rate, and cyclic fed-batch operations.     

A novel strategy on the high-cell-density cultivation of Candida utilis for the enhanced production of glutathione

Efficient glutathione production by high-cell-density cultivation of Candida utilis was investigated. A series of batch glutathione fermentations were carried out and the optimal initial glucose concentration was found to be about 26 g/L. Then, fed-batch fermentation under diverse feeding strategies was used to enhance glutathione production with a total glucose concentration of 150 g/L. Constant glucose feeding strategy cannot meet the requirement of cells at the late period of feeding, while exponential glucose feeding strategy cannot satisfy the needs of cells at the beginning of feeding. Based on the results above, a polynomial glucose feeding strategy was developed to provide enough glucose for cells along with the cultivation, under which both the cell and glutathione productivity were satisfactorily improved. Furthermore, fed-batch fermentation under this strategy with a total glucose concentration of 200 g/L was successfully performed, the dry cell weight and glutathione concentration reached 91.2 g/L and 825 mg/L, respectively.     

葡萄糖作辅助碳源对klebsiella pneumoniae发酵生产1,3-丙二醇的影响

在Klebsiella pneumoniae发酵甘油生产1,3 丙二醇的种子培养及发酵实验中,考察了葡萄糖作辅助碳源对菌体生长及1,3 丙二醇生成的影响. 结果表明,在种子培养期,以葡萄糖和甘油为混合碳源可缩短种子培养周期;在批次发酵和流加发酵中,葡萄糖作辅助碳源可使1,3 丙二醇产率及得率明显提高,但不同的葡萄糖加入方式对产率及得率促进的效果不同. 在初始发酵培养基中添加5 g/L葡萄糖、并在4 40 h进行葡萄糖与甘油混合液连续流加的条件下,1,3 丙二醇浓度、产率及得率较单一甘油为底物的流加发酵结果分别提高41.2 , 38.6 和8.3 .     

具有基质抑制的流加反应器计算机模拟和优化

以单位时间内获得尽可能多的细胞生物量为目标函数,建立了流加反应器最优化模型。通过引入哈密尔顿函数和庞特雅金最小值原理,求解了这个最优化模型。此外,以安德鲁方程为例,还对几种特殊的流加反应器进行了计算机模拟,并给出了最优化计算结果。     

基于pH恒定补糖的生物基丁二酸高效合成

通过确定大肠杆菌在产酸阶段葡萄糖的消耗与酸中和剂碳酸钠间的定量关系,建立了pH恒定补糖策略,能够使发酵液中葡萄糖浓度维持在稳定水平。与分批发酵相比,采用pH恒定补糖且维持葡萄糖浓度在较低的水平对丁二酸的积累是有利的。当采用pH恒定补糖并控制葡萄糖质量浓度在10g/L时,丁二酸最终质量浓度达到57.6 g/L,生产效率达到1.15 g/(L·h)。     

Enhanced production of glycyrrhetic acid 3-O-mono-β-D-glucuronide by fed-batch fermentation using p H and dissolved oxygen as feedback parameters

Glycyrrhetic acid 3-O-mono-β-D-glucuronide(GAMG), the major functional ingredient in licorice, has widespread applications in food, pharmacy and cosmetics industry. The production of GAMG through Penicillium purpurogenum Li-3 cultivation was for the first time performed through both batch and fed-batch processes in bioreactors. In batch process, under optimal conditions(p H 5.0, temperature 32 °C, agitation speed 100 r·min-1), 3.55 g·L-1GAMG was obtained in a 2.5 L fermentor. To further enhance GAMG production, a fine fed-batch process was developed by using p H and DO as feedback parameters. Starting from 48 h, 100 ml 90 g·L-1substrate Glycyrrhizin(GL) was fed each time when p H increased to above 5.0 and DO was increased to above 80%. This strategy can significantly enhance GAMG production: the achieved GL conversion was 95.34% with GAMG yield of 95.15%, and GAMG concentration was 16.62 g·L-1which was 5 times higher than that of batch. Then, a two-step separation strategy was established to separate GAMG from fermentation broth by crude extraction of 15 ml column packed with D101 resin followed by fine purification with preparative C18 chromatography. The obtained GAMG purity was 95.79%. This study provides a new insight into the industrial bioprocess of high-level GAMG production.