Preservation of industrially important cultures:
The isolation of appropriate organisms for a commercial procedure might be a long and very expensive procedure and it is thus necessary that it retains the desirable characteristics which led to its selection. Also, the culture employed to initiate industrial fermentation should be viable and free from contamination. Therefore, industrial cultures should be stored in such a way as to remove genetic change, protect against contamination and retain the viability. There are numerous techniques adopted to preserve microbial cultures. They are storage on agar slopes, storage beneath liquid nitrogen, dried cultures & lyophilization.
1) Storage on agar slopes:
Culture grown on agar slopes might be stored in a refrigerated (5oC) or a deep freezer and sub-cultured at around 6-monthly intervals. The time of subculture might be extended to 1 year when the slopes are covered with sterile medicinal grade mineral oil. This is the easiest and common technique of maintaining microbial cultures.
2) Storage under liquid nitrogen:The metabolic actions of microorganisms might be reduced significantly by storage at the very low temperature (i.e., -150oC to -196oC) that might be achieved by employing liquid nitrogen refrigerator. This approach is the most universally applicable of all preservation techniques. Fungi, viruses, bacteriophage, algae and yeasts have been successfully preserved in this technique.
3) Dried cultures:
Dried soil cultures have been employed broadly for culture preservation mainly for sporulating mycelial organisms. Moist, sterile soil might be inoculated with a culture and incubated for some days for some growth to happen and then permitted to dry at room temperature for around 2 weeks. The dry soil might be stored in a dry environment. This method has been employed extensively for the storage of fungal and actinomycetes culture.
4) Lyophilization:
Lyophilization or freeze-drying, includes the freezing of a culture followed by its drying under vacuum, that outcomes in the sublimation of the cell water. The method includes growing the culture to the maximum stationary stage and resuspending the cells in a protective medium like serum or sodium glutamate. Some drops of the suspension are transferred to an ampoule that is then frozen and subjected to a high vacuum till sublimation is complete, after which the ampoule is sealed. The ampoules might be stored in a refrigerator and the cells might stay viable for 10 years or more. Lyophilization is very well-situated for service culture collections since, once dried, the cultures require no further attention and the storage equipment is not expensive and reliable. By this technique, culture can be preserved for a big period.
Whichever method is employed for the preservation of industrial culture it is necessary to be certain of the quality of stocks. Every batch of newly preserved cultures must be routinely checked to make sure their quality.
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