固定化黑曲霉发酵玉米糖液生产柠檬酸的研究*

利用PVA复合凝胶包埋黑曲霉的孢子及菌丝体。固定后的细胞经过一系列预培养 ,用于发酵玉米生产柠檬酸。试验确定的固定化细胞摇瓶发酵生产柠檬酸最适条件为 :玉米糖液浓度 1 0Bx,培养温度 35℃ ,摇瓶转速 2 5 0r/min。经此条件发酵 64h ,柠檬酸产率最高达到 96g/L ,通常稳定在 90g/L左右。同时对柠檬酸连续批次发酵生产进行了初步研究 ,固定化黑曲霉可连续使用 8批次以上 ,其柠檬酸产量稳定在 86～ 92g/L之间 ,这为柠檬酸连续发酵提供了有利的保证 ,并探讨了有关的工艺技术条件     

固定化黑曲霉发酵玉米糖液生产柠檬酸的研究

利用PVA复合凝胶包埋黑曲霉的孢子及菌丝体。固定后的细胞经过一系列预培养,用于发酵玉米生产柠檬酸。试验确定的固定化细胞摇瓶发酵生产柠檬酸最适条件为：玉米糖液浓度10Bx,培养温度35℃,摇瓶转速250r/min。经此条件发酵64h,柠檬酸产率最高达到96g/L,通常稳定在90g/L左右。同时对柠檬酸连续批次发酵生产进行了初步研究,固定化黑曲霉可连续使用8批次以上,其柠檬酸产量稳定在86－92g/L之间,。这为柠檬酸连续发酵提供了有利的保证,并探讨了有关的工艺技术条件。     

聚氨酯固定化热带假丝酵母发酵木糖醇

固定在多孔聚氨酯载体中的热带假丝酵母(Candida tropicalis), 可有效地利用玉米芯半纤维素水解液生产木糖醇。在摇瓶条件下, 采用分批发酵方式, 确立了适宜的发酵工艺参数为: 接种量7%, 聚氨酯加入量1.0 g/100 mL, 温度30°C, 初始pH值6.0, 分段改变摇床转速进行溶氧调节, 其中0~24 h 为200 r/min; 24 h~46 h为140 r/min。聚氨酯固定化提高了菌体对发酵抑制物的耐受力, 固定化细胞密度高, 发酵性能稳定, 发酵产率和体积生产速率都有所提高。水解液未经脱色与离子交换便可转化成木糖醇, 大幅降低了成本, 显示了良好的应用前景。固定化细胞连续重复进行12批次21 d的发酵, 木糖醇得率平均为67.6%, 体积生产速率平均为1.92 g/(L·h)。     

聚氨酯泡沫固定化黑曲霉P-6021可同时产果胶酶和澄清果汁

以多孔聚氨酯泡沫(PUF)固定化黑曲霉P-6021可以实现菌丝体的摇瓶重复间歇发酵产酶,重复5个批次,菌体数目增多,产酶量上升,发酵液酶活达到664u/ml。以PUF固定化黑曲霉P-6021进行了同时产果胶酶和澄清苹果汁试验,以浑浊苹果汁基质发酵12h后, 产酶量可达到643u/ml,苹果汁基本澄清,透光率提高,粘度降低。表明固定化黑曲霉可以同时产果胶酶和澄清苹果汁,实现产酶与澄清过程的耦合。     

聚氨酯泡沫固定化黑曲霉P-6021可同时产果胶酶和澄清果汁*

以多孔聚氨酯泡沫(PUF)固定化黑曲霉P-6021可以实现菌丝体的摇瓶重复间歇发酵产酶,重复5个批次,菌体数目增多,产酶量上升,发酵液酶活达到664u/ml。以PUF固定化黑曲霉P-6021进行了同时产果胶酶和澄清苹果汁试验,以浑浊苹果汁基质发酵12h后, 产酶量可达到643u/ml,苹果汁基本澄清,透光率提高,粘度降低。表明固定化黑曲霉可以同时产果胶酶和澄清苹果汁,实现产酶与澄清过程的耦合。     

DA-F127水凝胶包埋固定化含腈水合酶细胞

腈水合酶是一类可催化腈类化合物转化生成相应酰胺类物质的酶。含腈水合酶的游离细胞催化水合反应存在酶容易失活、细胞无法重复利用、分离纯化困难等缺陷,细胞固定化技术可有效解决这些问题。为探索合适的固定化方法,以含腈水合酶的重组E.coli细胞为研究对象,以固定化酶活回收率和批次反应情况为评价指标,筛选比较了几种常用的包埋固定化方法。结果表明,DA-F127水凝胶包埋固定化细胞不仅具有较高的酶活回收率,而且稳定性也很好。对该方法进行了固定化条件和操作稳定性优化,当DA-F127浓度为15%、UV光源距离为20cm、光照时间为6min、菌体含量为20mg/g 固定化细胞时,酶活回收率为89.74%,并且可以催化9批次150g/L的3-氰基吡啶完成转化,第九批次转化率可达98.26%。与游离细胞催化过程相比,单位质量游离细胞的烟酰胺产量提高了12倍,具有良好的工业应用前景。     

DA-F127水凝胶包埋固定化含腈水合酶细胞

腈水合酶是一类可催化腈类化合物转化生成相应酰胺类物质的酶。含腈水合酶的游离细胞催化水合反应存在酶容易失活、细胞无法重复利用、分离纯化困难等缺陷,细胞固定化技术可有效解决这些问题。为探索合适的固定化方法,以含腈水合酶的重组E. coli细胞为研究对象,以固定化酶活回收率和批次反应情况为评价指标,筛选比较了几种常用的包埋固定化方法。结果表明,DA-F127水凝胶包埋固定化细胞不仅具有较高的酶活回收率,而且稳定性也很好。对该方法进行了固定化条件和操作稳定性优化,当DA-F127浓度为15%、UV光源距离为20cm、光照时间为6min、菌体含量为20mg/g固定化细胞时,酶活回收率为89. 74%,并且可以催化9批次150g/L的3-氰基吡啶完成转化,第九批次转化率可达98. 26%。与游离细胞催化过程相比,单位质量游离细胞的烟酰胺产量提高了12倍,具有良好的工业应用前景。     

聚氨酯泡沫固定化黑曲霉P—6021可同时产果胶酶和澄清果汁

以多孔聚氨酯泡沫（PUF）固定化黑曲霉P－6021可以实现菌丝体的摇瓶重复间歇发酵产酶,重复5个批次,菌体数目增多,产酶量上升,发酵液酶活达到664u/ml,以PUF固定化黑曲霉P－6021进行了同时产果胶酶和澄清苹果汁试验,以浑浊苹果汁基质发酵12h后,产酶量可达到643u/ml,苹果汁基本澄清,透光率提高,粘度降低,表明固定化黑曲霉可以同时产果胶酶和澄清苹果汁,实现产酶与澄清过程的耦合。     

固定化植物乳杆菌的制备及其发酵条件优化

采用海藻酸钠包埋植物乳杆菌并通过测定固定化细胞发酵清液的抑菌效果,优化得到的固定化最佳工艺条件为:海藻酸钠浓度为3%,CaCl2浓度为1.5%,菌悬液体积为3.5 mL(4.0×108 cfu/mL).固定化细胞重复发酵多批次效果良好.固定化细胞发酵条件优化结果表明:最适pH为7.0,最适温度为36℃,培养基中添加0....     

PVA共固定化双菌种发酵海藻酒的研究*

采用PVA为载体共固定化酿酒酵母和产香酵母发酵海藻,酿造海藻酒。对游离细胞与固定化细胞的分批发酵和连续发酵的动力学进行了研究并建立了相应的发酵动力学方程。实验结果表明：酿酒酵母和产香酵母二种菌种菌量的最佳配比为4：l,发酵温度20℃．共固定化细胞分批发酵和连续发酵凝胶粒的充填系数分别为0．25和0．5,游离混合细胞的发酵时间为7d。共固定化细胞连续发酵稀释速率0．12／h,其发酵时间为0.5d左右。经160d连续发酵实验,PVA固定化细胞粒子的机械强度良好。     

Removal of glucose from egg prior to spray drying by fermentation with immobilized yeast cells

Summary Baker's yeast cells (40% w/v) were immobilized in 2% Ca-alginate and were used in a batch process for the removal of glucose from egg melange by fermentation. Immobilized cells (10 g) could completely remove the glucose from 135 ml melange in 60 min. The immobilized preparation was repeatedly used in 30 batches without decrease in the activity.     

Preparation of lactose free milk by fermentation using immobilizedSaccharomyces fragilis

Summary Saccharomyces fragilis cells (40% w/v) were immobilized in 2% Ca-alginate and were used in a batch process for the removal of lactose from milk by fermentation. Immobilized cells (10 g) could completely desugarate 100 mL of milk in 3.5 h. The immobilized preparation was used repeatedly in 15 batches without decrease in the activity.     

Water reuse in the L-lysine fermentation process

L-Lysine is produced commercially by fermentation. As is typical for fermentation processes, a large amount of liquid waste is generated. To minimize the waste, which is mostly the broth effluent from the cation exchange column used for l-lysine recovery, we investigated a strategy of recycling a large fraction of this broth effluent to the subsequent fermentation. This was done on a labscale process with Corynebacterium glutamicum ATCC 21253 as the l-lysine-producing organism. Broth effluent from a fermentation in a defined medium was able to replace 75% of the water for the subsequent batch; this recycle ratio was maintained for three sequential batches without affecting cell mass and l-lysine production. Broth effluent was recycled at 50% recycle ratio in a fermentation in a complex medium containing beet molasses. The first recycle batch had an 8% lower final l-lysine level, but 8% higher maximum cell mass. In addition to reducing the volume of liquid waste, this recycle strategy has the additional advantage of utilizing the ammonium desorbed from the ion-exchange column as a nitrogen source in the recycle fermentation. The major problem of recycling the effluent from the complex medium was in the cation-exchange operation, where column capacity was 17% lower for the recycle batch. The loss of column capacity probably results from the buildup of cations competing with l-lysine for binding. (c) 1996 John Wiley & Sons, Inc.     

Broth recycle in a yeast fermentation

Fermentation is a water-intensive process requiring treatment of large amounts of effluent broth. It is desirable to increase the ratio of product produced to the volume of effluent by minimizing the discharge of effluent from the fermentation process. A study of recycling spent fermentation process. A study of recycling spent fermentation broth for the subsequent fermentation was carried out with Apiotrichum curvatum an oleaginous yeast, as the working culture. Spent broth from a defined medium was recycled t replace as much as 75% of the water and salts for subsequent batches and this was repeated for seven sequential batches without affecting cell mass and lipid production. A 64% vlume reduction of wastewater was achieved in this manner. However, when using whey permeate as the medium, lipid production dropped after three consecutive recycle operations at 50% recycle, and after two consecutive recycle operations at 75% and 100% recycle. Accumulation of ions in the broth appeared to be responsible for the inhibition. An ion exchange step was able to eliminate the ion buildup and restore fermentation performance. (c) 1994 John Wiley & Sons, Inc.     

High-level production of heterologous proteins using untreated cane molasses and corn steep liquor in Escherichia coli medium

To develop an economical industrial medium, untreated cane molasses (UCM) was tested as a carbon source for fermentation culturing of Escherichia coli. To test the industrial application of this medium, we chose a strain co-expressing a carbonyl reductase (PsCR) and a glucose dehydrogenase (BmGDH). Although corn steep liquor (CSL) could be used as an inexpensive nitrogen source to replace peptone, yeast extract could not be replaced in E. coli media. In a volume of 40 ml per 1-l flask, a cell concentration of optical density (OD₆₀₀) 15.1 and enzyme activities of 6.51 U/ml PsCR and 3.32 U/ml BmGDH were obtained in an optimized medium containing 25.66 g/l yeast extract, 3.88 g/l UCM, and 7.1% (v/v) CSL. When 3.88 g/l UCM was added to the medium at 6 h in a fed-batch process, the E. coli concentration increased to OD₆₀₀ of 24, and expression of both PsCR and BmGDH were twofold higher than that of a batch process. Recombinant cells from batch or fed-batch cultures were assayed for recombinant enzyme activity by testing the reduction of ethyl 4-chloro-3-oxobutanoate to ethyl (S)-4-chloro-3-hydroxybutanoate (CHBE). Compared to cells from batch cultures, fed-batch cultured cells showed higher recombinant enzyme expression, producing 560 mM CHBE in the organic phase with a molar yield of 92% and an optical purity of the (S)-isomer of >99% enantiomeric excess.     

Repeated batch production of citric acid from sugarcane molasses using recycled solid-state surface culture ofAspergillus niger

Summary The recycled solid-state surface fermentation (SSF) culture ofAspergillus niger KCU520 was used for repeated batch production of citric acid from sugarcane molasses. The rate of citric acid production was doubled, reducing the fermentation time to half, compared to the normal single cycle batch submerged or surface fermentation process. About 80% sugar was converted to citric acid in five-day batch fermentation and three batches were carried out with the same fungal mat without any significant loss of productivity.     

三相鼓泡塔生物反应器培养云芝菌合成漆酶

为了提高云芝菌发酵生产漆酶的效率,提出了一种利用自絮凝菌丝球在三相鼓泡塔生物反应器中重复分批发酵产漆酶的新方法。在优化后的产酶条件下,考察维生素C的添加浓度对漆酶活力的影响,并通过在培养基中添加维生素C进行漆酶多批次培养。研究结果表明,维生素C的添加浓度为1.50mmol/L时,可使漆酶活力提高2.6倍。连续进行了10批培养,每批最大漆酶的活力均在1000 U/mL以上,最高酶活出现在第五批为1919.6 U/mL,总培养时间达25 d。此方法所得漆酶对染料靛蓝具有明显的脱色降解作用,在介体1-羟基苯并三唑（HBT）用量0.10%,漆酶用量100 U/L条件下作用40 min时,靛蓝脱色率达到96.7%。该方法采用的三相鼓泡塔生物反应器性能稳定、菌丝球可重复使用,该方法有利于漆酶的高效、规模化生产。     

Semi-continuous xylose-to-xylitol bioconversion by Ca-alginate entrapped yeast cells in a stirred tank reactor

Candida guilliermondii FTI 20037 cells were entrapped in Ca-alginate beads and used for xylose-to-xylitol bioconversions during five successive batches in a stirred tank reactor. Supplemented sugarcane bagasse hemicellulosic hydrolysate was used as the fermentation medium. The average volume of the Ca-alginate beads was reduced by about 30% after the 600 h taken to perform the five bioconversion cycles, thus demonstrating physical instability under the conditions prevailing in the reactor vessel. In spite of this, almost steady bioconversion rates and yields were observed along the repeated batches. In average values, a production of 51.6 g l(-1), a productivity of 0.43 g l(-1 )h(-1) and a yield of 0.71 g g(-1) were attained in each batch, variation coefficients being smaller than 10%.     

Production of citric acid using its extraction wastewater treated by anaerobic digestion and ion exchange in an integrated citric acid-methane fermentation process

In order to solve the problem of extraction wastewater pollution in citric acid industry, an integrated citric acid-methane fermentation process is proposed in this study. Extraction wastewater was treated by mesophilic anaerobic digestion and then used to make mash for the next batch of citric acid fermentation. The recycling process was done for seven batches. Citric acid production (82.4 g/L on average) decreased by 34.1?% in the recycling batches (2nd–7th) compared with the first batch. And the residual reducing sugar exceeded 40 g/L on average in the recycling batches. Pigment substances, acetic acid, ammonium, and metal ions in anaerobic digestion effluent (ADE) were considered to be the inhibitors, and their effects on the fermentation were studied. Results indicated that ammonium, Na⁺ and K⁺ in the ADE significantly inhibited citric acid fermentation. Therefore, the ADE was treated by acidic cation exchange resin prior to reuse to make mash for citric acid fermentation. The recycling process was performed for ten batches, and citric acid productions in the recycling batches were 126.6 g/L on average, increasing by 1.7 % compared with the first batch. This process could eliminate extraction wastewater discharge and reduce water resource consumption.     

丙酸积累对薛氏丙酸杆菌生长及产酸的影响*

报道丙酸积累对维生素B12产生菌Propionibacterium shermanii生长及丙酸产生的影响,在初糖浓度6％,pH6.5的批次发酵条件下,测定了该菌的耗糖、产酸和茵体生长曲线。发酵24h后,培养基中添加1％、3％和6％的丙酸,发酵结束时菌体干重只有对照的75．2％、65．4％和52.9％,产酸是对照的79.3％、69．2％和39．3％。加入6％的丙酸不能完全抑制耗糖和产酸。部分解除丙酸抑制可使菌体干重增加60％。