Diacetyl production by immobilized citrate-positiveLactococcus lactis subsp.lactis 3022 in the fibrous Ca-alginate gel

Summary Diacetyl production by (Citr^*)Lactococcus lactis subsp.lactis 3022 was found to be an oxygen-dependent reaction. The diacetyl production by the cells immobilized in conventional Ca-alginate gel beads (Diameter: 3 mm) was lower than that of the cells immobilized in Ca-alginate gel fibers (Diameter: 0.2 mm), probably because oxygen transfer to the immobilized cells is better in gel fibers than in gel beads.     

D-malic acid production from DL-malic acid by enantiospecific assimilation with Acinetobacter lwofii

Summary Acinetobacter lwofii ATCC 9036 assimilated L-malic acid eantiospecifically and left D-malic acid when grown in a medium containing DL-malic acid. The optical purity of the D-malic acid isolated from the culture filtrate was 100%. When the organism was incubated at 26°C, 220 r.p.m. in a Erlenmeyer flask containing 100g/l of disodium maleate, L-malic acid was completely consumed during 7 days incubation and D-malic acid remained at the concentration of 35g/l.     

Diacetyl production by co-immobilized citrate-positiveLactococcus lactis subsp.lactis 3022 and homogenized bovin liver in alginate fibers with double gel layers

Summary Diacetyl production by (Citr⁺)Lactococcus lactis subsp.lactis 3022 cells immobilized in Ca-alginate fine fibers with single layer in the presence of catalase was three times higher than that in the absence of catalase. A co-immobilized culture system of the lactic acid bacterial cells (outer) and the homogenized bovine liver (inner layer) in Ca-alginate fibers with double gel layers was developed. The culture system gave high diacetyl productivity (30 mg/l) for ten repeated batch cultures.     

固定化细胞生产L—苹果酸动力学及两种酶反应器的比较

研究了产氨短杆菌ＭＡ－２,黄色短杆菌ＭＡ－３的固定化细胞在富马酸铵转化体系中生成Ｌ－苹果酸的动力学参数,同时比较了固定化细胞在填充床及连续机械搅拌反应器中酶转化反应的差异。研究结果表明：当转化率小于４０％时,酶反应在两种反应器所需的停留时间相当。随着转化率的提高,填充床反应器较连续机械搅拌反应器所需的停留时间短且不会因剪切力使固定化颗粒受到损伤,因此,在富马酸铵体系中用固定化酶生产Ｌ－苹果酸采用填     

Methane production from wastewaters by immobilized methanogenic bacteria

A methanogenic population was immobilized onto agar gel, polyacrylamide gel, and collagen membrane. Agar-gel-entrapped methanogenic microorganisms gave the highest activity. The optimum agar concentration was between 1.5 and 3% (w/v), and the optimum microbial content was 20 mg wet cells/g gel. The optimum conditions for methane production by immobilized whole cells were pH 7.0–7.5 and 37–45°C. The rate of methane production was initially 1.8 μmol/g gel/hr. Methane productivity was gradually increased and reached a steady state (4.5μmol/g gel/hr) after 25 days of incubation. The immobilized methanogenic microbial population continuously evolved methane over a 90 day period. No difference in methane productivity was observed after three months of storage at 5°C. Methane was also produced by immobilized whole cells under aerobic conditions. Furthermore, carbohydrates, such as glucose, in wastewater completely decomposed by immobilized whole cells.     

Screening of microorganisms having high fumarase activity and their immobilization with carrageenan

Summary To develop an efficient method for continuous production of L-malic acid from fumaric acid using immobilized microbial cells, screening of microorganisms having high fumarase activity was carried out and cultural conditions of selected microorganisms were investigated. As a result of screening microorganisms belonging to the genera Brevibacterium, Proteus, Pseudomonas, and Sarcina were found to produce fumarase in high levels. Among these microorganisms Brevibacterium ammoniagenes, B. flavum, Proteus vulgaris, and Pseudomonas fluorescens were further selected for their high fumarase levels in the cultivation on several media. These 4 microorganisms were entrapped into a k-carrageenan gel lattice, and the resultant immobilized B. flavum showed the highest fumarase activity and operational stability.Cultural conditions for the fumarase formation and the operational stability of fumarase activity of immobilized B. flavum are detailed. Productivity for L-malic acid using immobilized B. flavum with k-carrageenan was 2.3 fold of that using immobilized B. ammoniagenes with polyacrylamide.Presented at the Annual Meeting of the Agricultural Chemical Society of Japan, Nagoya, April 3, 1978     

Production of L-Malic Acid by Permeabilized Cells of Commercial Saccharomyces Sp. Strains

Of various yeasts tested in the conversion of fumaric to L-malic acid, Saccharomyces bayanus had the highest activity of fumarase. Cells permeabilized with 0.2% (w/v) CTAB for 5 min gave maximum enzyme activity. Under non-growth conditions, fumarase activity in the permeabilized cells was four times higher (271 U/g) than that of the intact cells (67 U/g). The proposed mathematical model for the batch production of L-malic acid was validated at different initial fumaric acid concentrations. The average conversion of fumaric acid was up to 82% and gave 21, 40, 83 and 175 mM L-malic acid from respectively, 25, 50, 100 and 210 mM: fumaric acid.     

Cultivation characteristics of immobilized Aspergillus oryzae for kojic acid production

Aspergillus oryzae in situ grown from spores entrapped in calcium alginate gel beads was used for the production of kojic acid. The immobilized cells in flask cultures produced kojic acid in a linear proportion while maintaining the stable metabolic activity for a prolonged production period. Kojic acid was accumulated up to a high concentration of 83 g/L, at which the kojic acid began to crystallize, and, thus, the culture had to be replaced with fresh media for the next batch culture. The overall productivities of two consecutive cultivations were higher than that of free mycelial fermentation. However, the production rate of kojic acid by the immobilized cells was suddenly decreased with the appearance of central cavernae inside the immobilized gel beads after 12 days of the third batch cultivation.     

根瘤菌BK-20固定化细胞生产L(+)-酒石酸

顺式环氧琥珀酸水解酶(CESH)是根瘤菌BK-20生产L(+)-酒石酸的关键酶。为提高其生产效率和生产稳定性,首先优化根瘤菌BK-20的产酶条件,然后利用固定化细胞连续生产L(+)-酒石酸。结果显示,优化后游离细胞酶活达(3 498.0±142.6)U/g,较优化前提高643%。固定化细胞酶活达(2 817.2±226.7)U/g,其最适包埋剂、菌体浓度和凝胶浓度分别为海藻酸钠,10%(W/V)和1.5%(W/V)。固定化细胞连续反应10批后,其形状和酶活均无明显改变,单批次转化率达98%以上,具有良好的生产稳定性。     

Morphological development of Aspergillus niger immobilized in Ca-alginate and K-carrageenan

Summary The morphological development of citric acid producing Aspergillus niger immobilized in Ca-alginate and K-carrageenan was studied. The fungus normally developed a dense mycelium layer below and on the gel bead surfaces so that substrate and oxygen in this area had direct contact with mycelia. By this way mycelia are not only immobilized by entrapment but also in a pellet-like matter.Limitation of the nitrogen source induces a more interior mycelium growth, and outgrowing of free cells was minimized. In sucrose media no effect on the particle stability was observed whereas the application of potassium acetate as substrate caused the complete dissolving of the matrices.     

Morphological examination of immobilized Streptomyces aureofaciens during chlortetracycline fermentation

Summary Immobilized spores of Streptomyces aureofaciens ATCC 10762 were grown in situ and formed micropellets under the gel surface. The latter was covered with a membrane-like coat of alginate material, while the bead interior was almost completely free of mycelial growth.High spore concentrations caused a decrease in antibiotic production which might be correlated with the morphological development of cells in the gel.Scanning electron micrographs showed the morphological development of the immobilized cells.     

碳源对固定化细胞生产柠檬酸的影响

柠檬酸是利用微生物代谢生产的一种极为重要的有机酸．广泛应用于食品、饮料、化工、冶金、印染等各个领域。在国外,近10年来,利用固定化细胞生产柠檬酸已获得较广泛的研究^〔1－6〕,国内也有学者指出,柠檬酸发酵的趋向是利用固定化细胞进行连续化生产⑺。而国内这方面的研究报道很少〔8,9〕。我们利用海藻酸钙凝胶包埋固定化黑曲霉细胞生产柠檬酸．探讨了碳源种类及其浓度对固定化细胞生产柠檬酸的影响。现将结果报道如下。     

Continuous ethanol production by immobilized cells of Zymomonas mobilis

Summary Studies have been carried out with a highly productive strain of Zymomonas mobilis in an immobilized cell reactor using both Ca alginate and -carrageenan as supporting matrices. Productivities above 50 g/l/h have been found at ethanol concentrations in excess of 60 g/l. With immobilized cells of Z. mobilis, there was a decline of approximately 30s% in activity after 800 h operation.     

Enhanced rate of ethanol production from D-xylose using recycled or immobilized cells ofPachysolen tannophilus

Summary Enhanced rates of ethanol production byPachysolen tannophilus from D-xylose were obtained by performing the fermentation with recycled cells in suspension culture or immobilized in a Ca-alginate gel. Fermentation under these conditions did not require aeration. Increasing temperature from 30 to 37°C enhanced the amount of ethanol produced in 24 hours from the recycled or the immobilized cells.Issued as National Research Council of Canada Publication Number 19475.     

Kinetics of repeated batch production of ethanol by immobilized growing yeast cells

Summary Kinetic and yield parameters for growth and ethanol production from sucrose (100 g/l) bySaccharomyces cerevisia entrapped in K-carrageenan and calcium alginate were identical to those of free cells. Cell leakage was minimum with calcium alginate gel. For the sixth batch, 4.51 g/lh ethanol productivity (94% conversion of sucrose) was obtained; 60.5 g/l of ethanol was obtained from 200 g/l sucrose with 83.2% conversion, indicating inhibition effects.     

Vertical Rotating Immobilized Cell Reactor of the bacteriumZymomonas mobilis for stable long-term continuous ethanol production

Summary Vertical Rotating Immobilized Cell Reactor was designed and built for glucose conversion into ethanol. Immobilized biomass units withZ. mobilis cells attached into polyurethane foam discs were fixed along a rotating shaft inside the bioreactor. The effect of rotation speed on the concentration of immobilized biomass was studied. Stability of the bioreactor over long-term operation was dependent on the concentration of the immobilized biomass. With fermentation carried out at 6 rpm a constant active immobilized cell concentration of only 34.5 g/l was maintained and used to convert up to 140 g glucose/l into more than 70 g ethanol/l with a volumetric ethanol productivity of 63 g/l/h.     

Proteomic analysis of calcium alginate-immobilized Saccharomyces cerevisiae under high-gravity fermentation conditions

Saccharomyces cerevisiae KAY446 cells immobilized in calcium alginate gel, and supplemented with additional amino acids, were successfully used in enhancing ethanol production. This combination succeeded in improving the ethanol yield and reducing the fermentation time. The ethanol yield under these conditions was 0.40 g of ethanol/g of glucose, with a final ethanol concentration of 118 g/L after 72 h. This is compared to yields with immobilized cells alone of 0.35 g of ethanol/g of glucose and freely suspended cells with no amino acid supplementation of 0.30 g of ethanol/g of glucose, under the same VHG conditions. The maximum specific ethanol production rates were 0.98, 0.73, and 0.61 g (g dry weight) (-1) h (-1) for immobilized cells under VHG conditions with and without amino acid supplementation and free cells, respectively. A proteomic analysis showed significant stimulation of many pathways during fermentation under these conditions, including the Ras/cAMP, glycolysis, starch, and sucrose pathways, amino acids biosynthesis, and aminoacyl-tRNA synthetases. The upregulation of ribosomal, heat-shock proteins and proteins involved in cell viability confirmed that protein biosynthesis was accelerated and revealed likely mechanisms for improving cellular viability.     

Production of ethanol by adsorbed yeast cells

Summary Various ion exchange resins were tested for their ability to adsorb cells of Saccharomyces cerivisiae with the ultimate intention of developing a packed bed immobilized cell reactor for the continuous production of ethanol. The resins varied greatly in their ability to adsorb cells - the least effective resins retained less than 1 mg S. cerivisiae cells (dry weight)/g of resin (dry weight), and the most effective, 130–140 mg cells/g of resin. A column reactor packed with adsorbed yeast cells was operated continuously for over 200 hours using a 12% (w/v) glucose medium at dilution rates of 1.1 h^-1 and 1.44 h^-1 (based on void volume). High ethanol productivities of 53.1 and 62.0 g ethanol/l-h were obtained.     

Enhanced yeast immobilization by nutrient starvation

Saccharomyces uvarum NRRL Y1347 cells were immobilized in a porous support. Cell loadings of up to 600 mg dry cell/g support or 70 mg dry cell/cm³ support were obtained. Starvation in a marine environment increased the adhesion strength of immobilized cells.     

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.