发展我国大规模细胞培养生物反应器装备制造业

生物反应器在生物工程装备制造业的产业化方面具有极其重要的意义。在分析了生物反应器产业的产品特点后,对我国生物反应器产业的现状和今后发展作了讨论。对以代谢流分析为核心的生物反应器、带pH测量与补料控制的摇床、动物细胞大规模培养生物反应器研制、生物反应器中试系统设计、大型生物反应器设计与制造技术研究等几个重要开发内容进行深入讨论,最后对细胞过程的系统生物学研究与生物反应器作了展望。     

植物细胞大规模培养生物反应器研制概况

本文对植物细胞培养中使用的搅拌式生物反应器,气升式生物反应器,固定化细胞生物反应器,光照培养生物反应器和其它新型生物反应器装置进行了全面的评述。     

植物细胞培养生物反应器的种类特点及展望

生物反应器技术应用于植物细胞培养既可以打破环境条件的限制,又有助于生产过程的人为调控,为植物细胞大规模培养或工厂化直接生产植物细胞有用代谢产物创造了条件,是当前植物细胞培养工作的研究热点。在介绍植物细胞培养特点的基础上,对适用于植物细胞培养的各类生物反应器(搅拌式生物反应器、非搅拌式生物反应器、用于植物细胞固定化培养的生物反应器、光生物反应器以及一次性培养生物反应器)的原理、优缺点等进行比较分析,最后提出了植物细胞培养生物反应器研究的发展方向,以期为植物细胞培养生物反应器的选择及改良提供参考。     

植物细胞大规模培养生物反应器研制概况

本文对植物细胞培养中使用的搅拌式生物反应器,气升式生物反应器,固定化细胞生物反应器,光照培养生物反应器和其它新型生物反应器装置进行了全面的评述。     

新型生物反应器结构研究进展

生物反应器是生物工程的核心设备,其结构的合理性直接决定反应器生物加工的效率。生物反应器的研究一直是生物工程的核心问题之一。随着青霉素的工业化生产,机械搅拌式生物反应器应运而生,此后,随着动植物细胞培养,高等真菌培养,藻类培养等生物过程的发展,人们相应开发了大量的生物反应器,其中以机械搅拌式生物反应器和气升式样生物反应器尤为突出,本文总结了近年来文献报道的新型生物反应器,主要阐述了机械搅拌式和气升式两类生物反应器结构的研究进展,对目前国内外报道的11种新型反应器典型结构进行了总结与分析。     

生物反应器及其研究技术进展

阐述了生物反应器设计、放大的新理念及关键技术发展,并在此基础上综述了应用于生物技术产品生产的生物反应器的主要发展趋势,包括以代谢流分析为核心的生物反应器系统、基于计算流体力学模拟技术的传统发酵罐改良、微型生物反应器、动物细胞反应器和酶反应器。     

封闭式光生物反应器研究进展

国际上80～90年代,封闭式光生物反应器是微藻生物技术的重要研究热点,也是微藻生物技术产业化的关键技术之一。本文较全面地介绍了用于微藻大规模培养的封闭式光生物反应器研究现状。将封闭式光生物反应器分为柱式、管式、板式和光导纤维反应器等类型。工业放大前景的管式和板式光生物反应器采取了典型个案分析的方法,列表比较了典型反应器的主要技术参数,并对它们的技术发展趋势进行了归纳总结。     

管道式光生物反应器的设计和性能

该文概述了管道式光生物反应器在设计上对性能的要求,对影响光生物反应器培养效率的各种生长条件如光能利用效率、CO2利用效率、环境温度、溶解氧等问题进行了探讨,指出高效并可自动调节的藻液循环混合系统对于高密度海藻培养是非常重要的,提出了能否自动清洗光生物反应器内壁是判断光生物反应器是否可用于工业化生产的关键。     

VERO细胞生物反应器放大培养初探

目的:研究用生物反应器放大进行Vero细胞微载体培养,实现生物反应器之间Veto细胞放大培养.方法:5L微载体生物反应器以10g/L微载体浓度培养Vero细胞,96h时经漂洗、消化、接种于30L微载体生物反应器,实现放大后的30L微载体生物反应器细胞怏速增殖,期间对不同时期的微载体细胞进行细胞计数、细胞代谢分析和形态观察.结果:5L生物反应器细胞经过96h灌注培养,平均细胞密度达到7.81×10~6cells/mL.5L微载体细胞放大到30L微载体生物反应器,平均细胞收获率为32.3%;放大到30L生物反应器后经过144h培养,细胞密度达到9.19×10~6cells/mL;放大后的细胞代谢途径依然以葡萄糖氧化代谢乳酸为主.结论:生物反应器由5L到30L进行Veto细胞放大培养是可行的.     

转基因动物生物反应器研究及产业化进程

<正>概述了生物反应器的概念与分类,介绍并比较了不同动物生物反应器的研究现状和发展前景,提出乳腺是生产结构和功能复杂的重组蛋白的最佳生物反应器,具有广泛的应用潜力,可在生产新型牛奶和药用珍稀蛋白等方面为人类做出巨大的贡献。同时指出了包括乳腺反应器在内的动物生物反应器存在的问题,对乳腺生物反应器的应用前景进行了展望。     

生物热对传统固态发酵菌群演替及其代谢影响的研究进展

解析传统固态发酵中产生的生物热对微生物菌群代谢的影响,是认识发酵机制、调控发酵过程、保证发酵效率的关键之一。固态发酵过程中,微生物菌群代谢活动所产生的生物热及传热效率低等问题引起微环境温度升高,进而影响微生物的生长与代谢。然而,关于传统固态发酵微生物受生物热的影响及其适应机制仍不明晰。因此,本文以传统固态发酵体系为研究对象,阐述持续生物热介导的高温对固态发酵过程中微生物群落演替和代谢功能的影响,并提出复杂群落中具有多层次调控微生物代谢以适应高温环境的方式,主要从微生物群体与个体层面介绍可能存在的耐热机制。了解生物热对传统固态发酵微生物的影响及潜在的耐热机制,有助于靶向调控发酵过程、强化高温发酵等,以满足未来的工业化需求。     

牲畜粪便与麦秆混合厌氧发酵的产气量、发酵时间及最优温度

探索牲畜粪便与作物秸秆混合发酵的产气量和发酵时间与发酵温度之间的关系,是解决农村户用沼气原料选择、确定最优发酵温度和提高农作物秸秆资源化利用效率的关键.采用可控型恒温发酵装置,以猪粪、牛粪和麦秆作为发酵原料,以常温厌氧发酵池的底物为接种物,在总固体（total solid,TS）质量分数为8%的条件下进行批量试验,研究了混合发酵的产气量、发酵时间及最优温度.结果表明：粪便与麦秆混合发酵明显提高了原料的产气效率,其中猪粪与麦秆混合发酵的累积产气量比猪粪作为单一发酵原料高24倍,而牛粪与麦杆混合发酵的累积产气量与单一牛粪无显著差异.猪粪、牛粪与麦秆混合发酵的最优温度均在30 ℃以上,发酵时间在60 d左右.厌氧发酵的发酵时间不总是随着温度的升高而缩短,单一以温度来断定厌氧发酵时间的长短是不可行的.     

Novel Method of Lactic Acid Production by Electrodialysis Fermentation

In lactic acid fermentation by Lactobacillus delbrueckii, the produced lactic acid affected the lactic acid productivity. Therefore, for the purpose of alleviating this inhibitory effect, an electrodialysis fermentation method which can continuously remove produced lactic acid from the fermentation broth was applied to this fermentation process. As a result, the continuation of fermentation activity was obtained, and the productivity was three times higher than in non-pH-controlled fermentation. In electrodialysis fermentation, the amount of produced lactic acid was 82.2 g/liter, which was about 5.5 times greater than that produced in non-pH-controlled fermentation. It was concluded that these good results were obtained on account of alleviating the lactic acid inhibitory effect by electrodialysis fermentation. However, the fouling of anion-exchange membranes by cells was observed in electrodialysis fermentation.     

Multi-energy metabolic mechanisms of the fungus Fusarium oxysporum in low oxygen environments

The fungus Fusarium oxysporum produces energy under hypoxic and anoxic conditions by denitrification (nitrate respiration) and ammonia fermentation respectively. Here we found that glucose repressed both of these metabolisms, whereas it supported another anoxic metabolism, hetero-lactic acid fermentation. Ammonia fermentation occurred only after the glucose present in the medium was metabolized to ethanol via alcohol fermentation. During this transition, clear diauxic growth was observed. Glucose regulated the activity of the enzymes involved in ammonia fermentation, hetero-lactic acid fermentation, and denitrification. Highest cell growth was supported by hetero-lactic acid fermentation, followed by denitrification and ammonia fermentation. These results indicate that the energy metabolisms of F. oxysporum are dependent not only on environmental O(2) tension but also on the carbon source, and that ammonia fermentation is an adaptative mechanism acting physiologically as a secondary fermentative mechanism replacing the primary hetero-lactic acid fermentation.     

Production of Botrytis cinerea for potential introduction into a vineyard

Botrytis cinerea was produced in solid-phase fermentation, liquid fermentation and on potato dextrose agar. Stored products were evaluated for grape colonization in grape bioassays and in field trials, and for B. cinerea density using colony forming unit analyses and a nucleic-acid-based method. B. cinerea colony forming unit density was significantly correlated to the probability of successful grape colonization in grape bioassays (p-value=0.0002). Solid fermentation products could be stored longer than liquid fermentation and potato dextrose agar products. There was little difference in the rate of grape colonization in laboratory bioassays among solid-phase fermentation, liquid fermentation and plate culture products. Although the initial B. cinerea colonization rate of field grapes was slightly greater on vines treated with solid-phase fermentation and plate culture products compared to vines treated with product from liquid fermentation, there was no significant difference in final colonization between vines treated with solid-phase fermentation, liquid fermentation and plate culture products and untreated vines.     

燃料乙醇发酵技术研究进展

在分析木质纤维素类生物质制备燃料乙醇原理基础上,重点对燃料乙醇转化过程的发酵工艺进行了论述。目前乙醇发酵工艺主要包括直接发酵、分步糖化发酵、同步糖化发酵、同步糖化共发酵和联合生物加工技术等,对这几种技术的研究现状进行了分析并对其发展趋势进行了展望,通过基因工程构建高效发酵菌种的联合生物加工技术将是未来高效发酵工艺的发展趋势,旨在为有效提高发酵菌株的底物代谢能力,获得高的乙醇产量提供重要参考。     

微生物发酵青蒿叶和叶渣的研究

为扩大青蒿原料的应用途径,延伸青蒿产业链,对青蒿叶和叶渣进行发酵研究。拟开发可用于动物保健的青蒿来源的产品。采用微生物发酵青蒿及青蒿叶渣,检测枯草芽孢杆菌、酿酒酵母菌、植物乳杆菌等菌株发酵青蒿叶和叶渣后其粗蛋白、粗脂肪、粗纤维素以及青蒿素、青蒿乙素、双氢青蒿酸、青蒿酸含量变化。青蒿叶发酵产物及功效成分含量与对照组比较,酵母菌发酵后粗蛋白提高43.05%,植物乳杆菌发酵后粗脂肪提高106%,枯草芽孢杆菌发酵后粗纤维降低43.30%,黑曲霉菌发酵后青蒿素和青蒿乙素分别提高133.27%和88.06%,地衣芽孢杆菌发酵后青蒿酸提高21.49%,枯草芽孢杆菌发酵后双氢青蒿酸提高86.01%;青蒿叶渣发酵产物与对照组比较,植物乳杆菌发酵后粗脂肪提高87.73%,酿酒酵母菌发酵后粗蛋白提高85.30%,枯草芽孢杆菌发酵后粗纤维降低55.67%,枯草芽孢杆菌发酵后双氢青蒿酸提高71.91%,地衣芽孢杆菌发酵后青蒿乙素提高94.71%。微生物发酵青蒿叶和叶渣显著提高其功效成分含量,增加了探索青蒿叶和叶渣发酵产物作为动物保健品的可能性。     

Acetic Acid Production by an Electrodialysis Fermentation Method with a Computerized Control System

In acetic acid fermentation by Acetobacter aceti, the acetic acid produced inhibits the production of acetic acid by this microorganism. To alleviate this inhibitory effect, we developed an electrodialysis fermentation method such that acetic acid is continuously removed from the broth. The fermentation unit has a computerized system for the control of the pH and the concentration of ethanol in the fermentation broth. The electrodialysis fermentation system resulted in improved cell growth and higher productivity over an extended period; the productivity exceeded that from non-pH-controlled fermentation. During electrodialysis fermentation in our system, 97.6 g of acetic acid was produced from 86.0 g of ethanol; the amount of acetic acid was about 2.4 times greater than that produced by non-pH-controlled fermentation (40.1 g of acetic acid produced from 33.8 g of ethanol). Maximum productivity of electrodialysis fermentation in our system was 2.13 g/h, a rate which was 1.35 times higher than that of non-pH-controlled fermentation (1.58 g/h).     

Performance comparison of ethanol and butanol production in a continuous and closed-circulating fermentation system with membrane bioreactor

Since both ethanol and butanol fermentations are urgently developed processes with the biofuel-demand increasing, performance comparison of aerobic ethanol fermentation and anerobic butanol fermentation in a continuous and closed-circulating fermentation (CCCF) system was necessary to achieve their fermentation characteristics and further optimize the fermentation process. Fermentation and pervaporation parameters including the average cell concentration, glucose consumption rate, cumulated production concentration, product flux, and separation factor of ethanol fermentation were 11.45?g/L, 3.70?g/L/h, 655.83?g/L, 378.5?g/m²/h, and 4.83, respectively, the corresponding parameters of butanol fermentation were 2.19?g/L, 0.61?g/L/h, 28.03?g/L, 58.56?g/m²/h, and 10.62, respectively. Profiles of fermentation and pervaporation parameters indicated that the intensity and efficiency of ethanol fermentation was higher than butanol fermentation, but the stability of butanol fermentation was superior to ethanol fermentation. Although the two fermentation processes had different features, the performance indicated the application prospect of both ethanol and butanol production by the CCCF system.     

L-缬氨酸的发酵工艺研究

目的:以黄色短杆菌XV0505为生产菌株,探讨发酵培养基和发酵控制条件对L-缬氨酸的产量和糖酸转化率的影响。方法:应用单因素实验确定发酵的工艺条件;利用纸层析-色斑洗脱比色法测定发酵液中L-缬氨酸的含量。结果:在最优发酵条件下,通过10L罐流加发酵72h,产酸量可达53.4g/L,糖酸转化率为26.7%,分别比补料分批发酵提高11.9%和3.5%。结论:环境因子和发酵控制工艺对发酵生产L-缬氨酸具有重要影响。