Culture Management

Isolation, Preservation and Improvement

 Points to Consider for Isolation

 Nutritional Characteristics of the organism

 The medium, additives, any other chemical factors

 The optimum temperature of the organism

 Based on the production condition and the

indented use of the products

 The reaction of the organism with the equipment to

be employed
 Suitability of the organism to the type of
process to be used
 The stability of the organism and its amenability
to genetic manipulations
 The productivity of the organism
 Substrate to product rate
 Product per unit time
 The ease of product recovery from culture)
 SOURCES OF THE MICROORGANISMS

Culture collection Fast & Time saving

 Cheap
 Model systems to develop assay system &
standardization
 May be a weak producer of product

Natural Sources
 Slow and time consuming
 May be costlier
 Have opportunity to find a better producer
 CULTURE COLLECTIONS

 National Collection of Type Cultures (NCTC)-London UK

 National collections of Industrial & Marine Bacteria Ltd.

(NCIB, NCMB)–UK

 National Collection of Yeast Cultures (NCYC) – UK

 Collection of International Mycologica l Institute (IMI) –

UK

 American Type Culture Collection (ATCC)- Rockville- USA

 Deutsche Sammlung von Mikroorganismen and

Zelkulturen (DSM)- Germany
 Centraalbureau voor Schimmelcultures (CBS) –
Netherlands

 Czechoslovak Collection of Microorganism (CCM) – Czech

republic

 Collection of Nationale de Culture de Microorganisms

(CNCM)

 Japan Collection of Microorganisms (JCM)- Japan

 Culture Collection of the Institute for fermentation (IFO) –

Japan

 Microbial Type Culture Collection (MTCC) -

 ISOLATION STRATEGIES

 Selection pressure
 Culture conditions by means of substrate
 Selection of a taxon known to have a particular
properties in high frequency
 Streptomycetes or actinomycetes
 Now metagenomic approaches
  ISOLATION METHODS

 Enrichment using liquid media

 Batch liquid enrichment & continuous liquid
enrichment
 Enrichment using solid media
 Alkaline protease producing bacillus sp. & cellulase
producing organisms
ISOLATION , NOT USING SELECTION OF DESIRED
CHARACTERISTICS

 Enzyme assays
 Molecular biological methods
 Immuno assays
 Molecular Probes
 Reporter assays
 STORAGE OF MICROORGANISMS

 Solid Agar medium

 Slants
 Stabs
 Plates
 Liquid medium
 Liquid Nitrogen
 Dried form
 Dried with an inert substrate
 Freeze dried form
 PRECAUTIONS TO BE TAKEN

 Genomic Characterization
 Contamination
 Product production rates etc
 IMPROVEMENTS OF THE CULTURE
 Selection of varieties/strains
 Selection of natural mutants
 Artificial mutations
 Isolation of auxotophic mutants
 Genetic modifications
 Modification of permeability
 Modification of stability / solubility
 Protoplast fusion
 rDNA technology