Calcium-alginate-immobilized

Shen, B., Greenfield, P., and Reid, S., Calcium Alginate Immobilized Hybridomas Grown Using a Fluidized-Bed Perfusion System with a Protein-Free Medium, Cytotechnol., 14 109 (1994)... 

A collective anaerobic and aerobic treatment by immobilized microorganisms was first shown [66, 67] using calcium-alginate-immobilized co-cultures of a facultative anaerobic strain of Enterobacter cloacae. Reaction actually took place in the middle of the alginate beads. In these experiments, the reduced derivatives were oxidized in the outer parts of the alginate beads by a second aerobic strain (two different Alcaligenes species), which had the ability to oxidize 4-chloro-2-aminophenol. 

Hulst and co-workers obtained a value for the 02 uptake in both suspended and calcium alginate immobilized Daucus carota (carrot) cells [31]. This value was approximately 3 X 10 7 mol g v s-1. Nevertheless, the value of Hallsby s estimate of uptake of oxygen, 3 x 10-10 g v mol-1 s 1 for N. tabacum, will be used [32,33]. Hulsfs observation that specific consumption was similar for suspended and immobilized cells is assumed to apply [31]. 

During most of our studies with calcium alginate immobilized N. tabacum cells, the bioreactor columns contained about 0.04 g (dry weight) of cells of course, this value varied from test to test. It is reasonable to assume that the feed medium was saturated in 02. The 02 mole fraction, x, in the medium (at saturation) is given by Henry s Law [47] ... 

Both cell culture with a lipophilic extraction phase and with a polar extraction phase have been reported to be helpful for the accumulation and detection of secondary substances [7,8]. Plant cell cultures release lipophilic and volatile substances such as ethylene, ethanol, and acetaldehyde. The addition of a lipophilic phase to the culture medium can be used as a means of accumulating and detecting these substances. Maisch et al. [8] found that the addition of XAD-4 resin to Nicotiana tabacum cultures enhanced the production of phenolic secondary metabolites several times compared to the adsorbent-free control. Kim and Chang [9] reported in situ extraction for enhanced shikonin production by Lithospermum erythrorhizon. When n-hexadecane was added to the cultivation, higher specific shikonin productivity was obtained than that from the cultures of free cells without extraction. They also suggested that n-hexadecane addition at an early stage in calcium alginate immobilized cell cultures was effective for shikonin production. Most of the produced shikonin was dissolved in n-hexadecane, so it would reduce the costs for shikonin separation. 

Figure 9. Dependence of residual activity (effectiveness of immobilized cells) on particle size and cell loading for the production of gluconic acid from glucose with calcium alginate immobilized Acetobacter simplex cells.

Goksungur, Y. and Guven , U. (1999) Production of lactic acid from beet molasses by calcium alginate immobilized Lactobacillus delhrueddi IFO 3202. J. Chem. Technol. Biotechnol, 74, 131-136. 

Gontier E, Sangwan BS, Barbotin JN (1994) Effects of calcium, alginate, and calcium-alginate immobilization on growth and tropane alkaloid levels of a stable suspension cell line of Datura innoxia Mill. Plant Cell Rep 13(9) 533-536... 

Rickert DA, Glatz CE, Glatz BA (1998) Improved organic acid production by calcium alginate-immobilized propionibacteria. Enzyme Microb Technol 22 409-414... 

Calcium alginate [345-347] as well as K-carrageenan [348-350] has been used for immobilizing baker s yeast, and a sodium alginate/calcium chloride-immobilized baker s yeast has been used during the synthesis of d- and L-armentomycin [351]. This system has also been used for tiie reduction of several p-keto esters [352], and the antiselectivity as well as the yields was improved compared to reductions using classical conditions [217,353-355]. Calcium alginate-immobilized cells of S. cerevisiae were pretreated at 50 °C for 30 min, and tiien the cells showed an increased performance (80%) and an excellent selectivity (ee 99%) in the reduction of 3-chloropropiophenone... 

Calcium alginate-immobilized yeast cells were successfully used for the synthesis of small peptides [524]. Of special synthetic mterest seems to be the biohydrogenation of 2-substituted allyl alcohols 355 (Fig. 86) to afford enantioselectively the (/ )-2-methylal-kanols 356 [525,526]. For natural product synthesis several butenolides have been reduced very successfully with BY [527-529]. 

Pai S-L, Hsu Y-L, Chong N-M, Sheu C-S, Chen C-H (1995) Continuous degradation of phenol by Rhodococcus SR immobilized on granular activated carbon and in calcium alginate. Biores Technol 51 37 12... 

D-Pantolactone and L-pantolactone are used as chiral intermediates in chemical synthesis, whereas pantoic acid is used as a vitamin B2 complex. All can be obtained from racemic mixtures by consecutive enzymatic hydrolysis and extraction. Subsequently, the desired hydrolysed enantiomer is lactonized, extracted and crystallized (Figure 4.6). The nondesired enantiomer is reracemized and recycled into the plug-flow reactor [33,34]. Herewith, a conversion of 90-95% is reached, meaning that the resolution of racemic mixtures is an alternative to a possible chiral synthesis. The applied y-lactonase from Fusarium oxysporum in the form of resting whole cells immobilized in calcium alginate beads retains more than 90% of its initial activity even after 180 days of continuous use. The biotransformation yielding D-pantolactone in a fixed-bed reactor skips several steps here that are necessary in the chemical resolution. Hence, the illustrated process carried out by Fuji Chemical Industries Co., Ltd is an elegant way for resolution of racemic mixtures. 

Corynebacterium glutamicum (CGMCC No. 1464) cells immobilized in calcium alginate beads cross-linked with polyethenimine and glutaraldehyde have been employed for the production of nicotinamide from 3-cyanopyridine [21], The reaction was mn at 10-15 °C,... 

The decolorization potential of immobilized P. chrysosporium MTCC 787 for azo dyes Acid Orange, Acid Red 114, triphenylmethane dye Methyl Violet, diazoic dye Congo Red, vat dye Vat Magenta, thiazine dye Methylene Blue, and anthraqui-none Acid Green was demonstrated by Radha et al. [53]. Decolorization experiments were carried out with immobilized calcium alginate (Ca-ALG) beads of different sizes (2-6 mm). 

Draget KI, Myhre S, Ostgaard K. Plant protoplast immobilized in calcium alginate. A simple method of preparing fragile cells for transmission electron microscopy. Stain Technol 1988 63 159-163. 

Biotransformations of morphinan alkaloids have been reported for plant, fungal, and mammalian enzymatic systems with emphasis on rather specific reactions such as the reduction of ketones, N- and O-demethylation, and perox-idative transformations. Furuya et al. used immobilized tissue culture cells of Papaver somniferum to accomplish the selective reduction of codeinone (135) to codeine (136) (207) (Scheme 30). Suspension cultures of a well-established cell line of P. somniferum were grown for one week as a source of cell mass for immobilization in calcium alginate. The cells continued to live in the alginate matrix for 6 months maintaining their biological activity. The reduction of co-... 

When cells of Pseudomonas sp. OS-K-29 immobilized on calcium alginate were incubated in 80 1 of a synthetic medium containing 0.2 vol.% of racemic 2,3-DCP, optically pure (S)-2,3-DCP of 100% e.e. was obtained. The immobilized cells could be re-used for a series of continuous reactions, a bioreactor being used for 19 reactions over 50 days without any loss of activity (Fig. 3). Optically pure (i )-2,3-DCP (100% e.e.) was also isolated from the racemate by means of stereospecific assimilation by Alcaligenes sp. DS-K-S38. Highly pure (R)- and (S)-EP of 99.5% e.e. were prepared from (S)- and (i )-2,3-DCP by treatment with aqueous NaOH, respectively [20-221 (Fig. 2). 

While most appfications were performed in suspended cell cultures some authors showed that the application of NADH-dependent fluorescence monitoring is also possible in immobifized cell systems. Here the growth of Clostridium acetobutylicum and the Saccharomyces cerevisiae immobilized in different calcium alginate structures was studied. However, calibration of the culture fluorescence signal with the biomass concentration was not possible but qualitatively an increasing biomass also led to an increase in the fluorescence signals. 

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