isolation and identification of industrially important microbes

1
ISOLATION, IDENTIFICATION PRESERVATION OF
INDUSRTIAL MICRO-ORGANISMS.

• Presented By,
• PRANAV H.NAKHATE
• M.Tech Biochemical Engg.
• Amity University, Jaipur

2
POINTS TO BE DISCUSS
Microbial preservation
Identification of microbes
Isolation of microbes
Why microbes are important industrially ?
3
INTRODUCTION

• The microbe will have the last word.
  
  Louis Pasteur
• Industrial microbiology associates with the
  commercial exploitation of microbes for the
  benefit of mankind.
• These microbial products may have direct or
  indirect impact on the economics, environment and
  social parameters of the society.
• Microorganisms are used to create a wide variety
  of products and to assist in maintaining and
  improving the environment

4

• Microbes have wide scope in different areas. Some
  of them are,
• starter cultures in commercial food and beverage
  production.
• Products of alcoholic fermentation.
• Production of Enzymes and other industrial
  products.
• Production of Alternative fuels.
• Production of Pharmaceuticals, like
  antimicrobials, recombinant hormones, and other
  cell regulators.
• Pesticides and agricultural products
• Biosensors and bioreporters.

5
INDUSTRIAL STRATEGY FOR USAGE OF MICROBES

• Isolation of microbes from natural
  source.
• Identification of desired
  microbes.
• Characterisation of microbes
• Screening of desired
  microbes.
• Inoculum preparation
• Srain Improvement
• Fermentation

6
ISOLATION OF INDUSTRIAL IMPORTANT MICROBES

• Isolation is having mainly 2 strategies
• 1) from the environment by collecting samples of
  free living microorganism from natural habitats.
• 2) By sampling of microbes with desired
  characteristics from specific sites such as found
  among the natural micro flora.
• Hence, its important to isolate that specific
  microbe from the environment.
• The site for isolation of microbes is decided by
  the type of microbe one is interested in.

7
Sr. No. PRODUCT SITE
1 Thermophiles Hot spring, thermophilic vents, compost
2 Alkalophiles High salinity soils and wells
3 Acidophiles Pine growing sites, fruits
4 Osmophiles Seas and oceans
5 Yeast Wine yards
6 Phosphate solubilizers Agricultural soil, rhizosphere
7 Protease producers Dairy industry, meat processing units.
8 Enzymes like cellulase, pectinase, xylanase etc. Agricultural soil, paper and pulp industry, waste disposal area of fruit juice plants, grain market.
9 Biogas (methane) producers Gober-gas plants, effluent collection sites.
8
Cont. (Ref. 2)

• After sampling of the organism the next step is
  of enrichment.
• This increases the quantity of the desired
  organism prior to isolation and screening.
• Although numerous media have been devised for the
  growth of microbes, there is no single media that
  supports all kinds of microbes.
• E.g. A soil sample plated on nutrient agar (pH
  7.0), incubated at 37C for 24-48 hrs, will grow
  only aerobic mesophiles, but anaerobes,
  thermophiles, psychrophiles, alkalophiles and
  acidophiles will not grow

9
IDENTIFICATION OF INDUSTRIAL IMPORTANT MICROBES
(
Ref 3)

• After the isolation of bacteria, it is important
  to identify the specified bacteria.
• Identification is mostly done with the help of
  following characteristics
• 1) Morphology,
• 2) Selective and diagnostic media,
• 3) Cultural characteristics,
• 4) Additional recognized biochemical tests,
• 5) Profile of microbial stains, and
• 6) Rapid identification methods.

10

• 1) Morphology -
• It is the science dealing with the structures
  and forms of organisms.
• It exclusively trusts confidently upon a number
  of factors, such as shape, length and breadth,
  ends of organism, body arrangement, motility,
  forms spores, capsule

• 2) Selective and diagnostic media -
• It specifically favour the growth of particular
  microorganisms.
• Eg. MacConkeys agar medium for E. coli.
• Differential media, Enrichment media, and
  Characteristic media are types of this media.

11

• 3) Additional recognized biochemical tests -
• These tests are extensively used for
  identification purpose. These tests are,
• Carbohydrate (Sugar) Fermentation, Litmus Milk,
  Indole Production, Methyl Red Test MR-Test,
  Voges-Proskauer Test VP-Test, Citrate
  Utilization, etc.

• 4) Profile of microbial stains -
• microbes are extremely too small in size, hence
  to visualise them, staining is takes place, in
  which either cell interior or exterior part
  stains in colour.
• Different Staining technique includes, Gram
  staining, Differential staining, Capsule staining
  Endospore staining Flagellar staining.

12

• 5) Cultural characteristics -
• Based upon a broad spectrum of intensive and
  conclusive research carried out during the past
  few decade
• These characteristics are, Shape, Size, Elevation
  Margins, Surface, Edges ,Colour , Structure,
  Consistency, etc.

6) Rapid identification methods - These
includes advanced techniques like SEM, TEM.
13
CULTURE PRESERVATION

(Ref 4)

• The conventional microbiology is totally
  dependent on the availability of pure and stable
  cultures.
• serial transfer is often the first preservation
  technique used by microbiologists, but culture
  is prone to contamination by this.
• In order to prevent the loss of production of a
  desired metabolite and to preserve culture, a
  variety of culture preservation techniques have
  been developed.

14
1) Culture Transfer - To avoid contamination,
genetic change
2) Refrigeration from 0o to 5oC - short
term storage
3) Low Temperature Freezing - ultra low temp.
freezer (-80oC), liquid nitrogen (196oC)
4) Lyophilization freeze with dry ice and
acetone sublime off water (dries cells without
disruption) use of glycerol, or sucrose to
protect cells

• 5) Mineral Oil
• 6) Dry Spores

15
USES OF PROPER ISOLATION, IDENTIFICATION
PRESERVATION

• To Improve industrial production.
• To study proper morphology physiology of
  microbes.
• To understand microbial kinetics.
• Microbial biosensor.
• Biopesticides over Chemical pesticides.

16
REFERENCE

• Ref 1 - biotechnology. editor u. satyanarayanan.
  publishers books and allied (p) ltd. calcutta.
  india. 2005.
• Ref 2 Pharmaceutical Microbiology,sixth edition
  edited by w.b.hugo a.d.russell, professor of
  pharmaceutical microbiology university of wales
  cardiff, cardiff
• Ref 3 - Aamand, J., T. Ahl and E. Spieck. 2006.
  Monoclonal antibodies recognizing nitrite
  oxidoreductase of Nitrobacter hamburgensis, N.
  winogradskyi, and N. vulgaris. Appl. Environ.
  Microbiol.
• Ref 4 - Becker W.M., Kleinsmith L, and Hardin J.
  The World of the Cell, 4th ed., Benjamin
  Cummings, Redwood City, Calif., 2008.