Studies on immobilized fungal spores for microbial transformation of steroids: 11 alpha-Hydroxylation of progesterone with immobilized spores of Aspergillus ochraceus G8 on polyacrylamide gel and other matrices

Hydroxylation in the 11 alpha-position in the progesterone molecule employing immobilized spores of Aspergillus ochraceus strain No. G8 (CDRI catalogue No.) was achieved. For immobilization the activity of the spores was evaluated on a variety of matrices such as alginate beads, epoxy resin beads, polyacrylamide gel, and collagen. Spores entrapped in polyacrylamide gel were found to be the most active. Studies of various parameters, e.g. monomer content, cell loading capacity, optimum pH, temperature, and substrate concentration, were carried out on polyacrylamide gel. In polyacrylamide, the entrapped spores normal decay pattern, as indicated by loss of activity, was observed after four uses. At the end of 15 cycles, the residual activity was found to be 18% of the original. It was possible to regenerate the activity by incubating the preparation in a nutrient medium. The regenerated spores showed increasing rate of loss of activity upon recycling.     

Steroid hydroxylation using immobilized spores of Curvularia lunata germinated in situ

Summary Spores of Curvularia lunata were immobilized in polyacrylamide granules and in calcium alginate beads (2–3 mm in diam.). Germination of the spores, initiated by the addition of nutrients, resulted in an even distribution of mycelium throughout the beads after 48 h. Such beads were used for the conversion of cortexolone to cortisol by steroid-11-hydroxylation. In order to improve the steroid transforming ability several parameters were studied. It was found that preparations based on calcium alginate gave the best results.The possible merits of immobilizing spores rather than vegetative cells, followed by in situ germination are discussed also for other microorganisms and immobilization processes.     

A method for starting a fermentor with immobilized spores of the genus Clostridium aseptically

Summary A method has been developed to start a fermentor with immobilized Clostridium butylicum spores aseptically.     

The surface coats on spores

There are surface coats on the sporocytes and spores of some pteridophytes and bryophytes which will bind one or more of the stains generally used to demonstrate the presence of an acid mucopolysaccharide surface coat in animal cells, viz. Alcian blue, colloidal iron, lanthanum, thorium, silver, SO,-coriphosphine and phosphotungstic acid. This suggests that the composition of the coat substances in the ferns and bryophytes agrees with that in animal cells. It has been found that thorium-staining in the sporocyte and the coat of the young spore of one fern, Botrychium lunaria , can be abolished with neuraminidase, indicating that sialic acid is a principal component of these coats. The spore wall in the pteridophytes (and probably also in the bryophytes) is constructed within these mucopolysaccharide surface coats, and it is suggested that differences in wall form are attributable to qualitative, quantitative and functional differences of the coats.     

Immobilization of fungal spores on synthetic polymers

Adsorption of fungal spores on a synthetic polymer - High Density Polyethylene was successfully achieved using different pretreatments. Sonication of beads followed by ferric nitrate treatment or use of 0.1% tributyrin gave adsorption upto 46%. Use of dichloromethane as a solvent for sonication is recommended for its better performance in reuse studies (upto 5 times without much decrease in activity). 100 g of immobilized biocatabyst in a 7 L Fluidized Bed Bioreactor was found to perform better than shaker flask at a much lower power input.     

Immobilization of fungal spores on synthetic polymers

Adsorption of fungal spores on a synthetic polymer - High Density Polyethylene was successfully achieved using different pretreatments. Sonication of beads followed by ferric nitrate treatment or use of 0.1% tributyrin gave adsorption upto 46%. Use of dichloromethane as a solvent for sonication is recommended for its better performance in reuse studies (upto 5 times without much decrease in activity). 100 g of immobilized biocatabyst in a 7 L Fluidized Bed Bioreactor was found to perform better than shaker flask at a much lower power input.     

Studies on immobilized papain

Papain was covalently coupled to ZrO_2?coated porous glass by several different methods. These derivatives were characterized and their operational half-lives determined using casein substrate. Papain covalently coupled to the porous glass, previously converted to a carboxylic acid derivative, through amide linkage gave a 35 day operational half-life.     

Uninucleate spores of Phycomyces

Summary Under most culture conditions only 0.3% of the vegetative spores of Phycomyces blakesleeanus are uninucleate. On an acidified minimal medium, the uninucleate fraction can be raised up to 4.5% of the spores. The spore population can be fractionated in a gradient under gravity (1xg) yielding fractions that contain over 80% uninucleate spores. These uninucleate spores are fully viable. When the spores to be fractionated are obtained from a heterokaryotic mycelium, the uninucleate fraction produces homokaryotic mycelia.