Isolation of industrially important microorganisms from soil

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BACKGROUND

In nature, there are about a million different species of microbes. Fewer than 1% of all microbes on the planet have been examined. Only a small number of species—less than a hundred—are crucial for commercial application.

Soils, lakes, and river muds make excellent sources for isolating microorganisms. A gram of soil is thought to contain between 106 and 108 bacteria, 104 and 106 actinomycete spores, and 102 and 104 fungus spores.

Biotechnologists frequently choose isolating microbes from extremely harsh and unique habitats. This is done in the hopes that these strains may be able to create novel, industrially significant products. For this goal, several unique conditions, including frigid habitats, high elevations, deserts, deep seas, and petroleum fields, are continuously tested.

The latest metabolites produced by bacteria are still being sought after by industrial microbiologists.

In addition to a number of other chemicals, research is primarily focused on identifying chemotherapeutically significant products for the treatment of cancers, bacterial diseases (newer antibiotics against resistant strains), and viral infections (e.g. hormones, enzyme inhibitors).

Industrially important microorganisms from soil

The isolation of microorganisms is important for the development of the food sector as well as for the effective breakdown of dangerous substances and environmental contaminants.

The microbes can be isolated and evaluated directly for product production. Actually, the soil or water samples can be utilized directly or with the appropriate dilution for metabolite screening.

Metabolites produced by the bacteria can be screened on agar plates. For instance, if the test product is an antibiotic, the organism strains on the agar plates that inhibit the zones serve as the test system.

The inhibitory activity suggests that, the microbes may be producing some sort of antibiotic. And another illustration is the isolation of bacteria that make amylases. Amylase-producing organisms can be recognized and isolated by growing them on starch-containing agar plates and then staining the cultures with iodine.

REQUIREMENTS

Sterile spatula
Sterile bag
Sterile distilled water
0.8% saline
Centrifuge
Olive oil
Tween 20 or Tween 80
50 mM Sodium phosphate buffer (pH 7.0)
Acetone: ethanol solution (1:1)
0.05 M NaOH solution
Phenolphthalein indicator
Tributyrin agar medium (Peptone 0.5 g/L, Yeast extract 0.3 g/L, tributyrin 1 g/L, Agar 2%)
Tryptic soy broth medium (Tryptone 10 g/L, Yeast extract 5 g/L, NaCl 10 g/L, pH 7.5)

PROCEDURE

  1. Collect soil with the help of sterile spatula and stored in sterile bags. [2]
  2. 1 gm of soil sample dissolve in 10 ml of sterile distilled water.
  3. From the above solution prepare serial dilution using the sterile distilled water or 0.8% saline.
  4. 100 µl of each dilution was spread on the tributyrin agar plates, and incubated at 37˚C for 24 hr. [1]
  5. Observe for lipolytic activity by the formation of hydrolysis zones around the bacterial colonies.
  6. Selected bacterial strains morphologically and biologically characterized by using the Bergey’s manual system.
  7. Prepare the tryptic soy broth medium for the lipase production and sterilize it.
  8. Inoculate it with the isolated bacterial strain, and incubate at 37˚C for 24 hr with constant shaking.
  9. Centrifuge the medium to obtain cell-free supernatant at 5000 rpm for 15 min at 4˚C.
  10. Collect the supernatant, and used to determine the lipolytic activity
  11. Lipolytic activity measured by the titrimetric method using olive oil as substrate, 10% olive oil was emulsified with 5% Tween 20 or Tween 80 in 50mM sodium phosphate buffer (pH 7.0). [1]
  12. 100 µl cell free supernatant added to the emulsion and incubate at 37˚C for 15-20 min, the reaction stopped and fatty acid were extracted by addition of 1.0 ml of acetone: ethanol solution (1:1).
  13. The amount of fatty acid liberated estimated by titrating with 0.05M NaOH using phenolphthalein as indicator.
Isolation of industrially important microorganisms from soil
Isolation of industrially important microorganisms from soil

CONCLUSION

The lipase-producing microorganism was isolated from the soil, its morphological and biochemical characterization was done by using Bergey’s manual system, and its lipolytic activity was measured by using the titration method.

REFERENCES

  1. Kalpana Sagar, Yasir Bashir, Mayur M Phukan, B. K. Konwar, “Isolation of Lipolytic Bacteria from Waste Contaminated Soil: A Study with Regard to Process Optimization for Lipase”, INTERNATIONAL JOURNAL OF SCIENTIFIC & TECHNOLOGY RESEARCH, VOLUME 2, ISSUE 10, OCTOBER 2013.
  2. F Soundra Josephine, Ramya V S, Neelam Devi, Suresh Babu Ganapa, Siddalingeshwara K. G. Venugopal. N and Vishwanatha T, “Isolation, production and characterization of protease from Bacillus Sp isolated from soil sample”, J. Microbiol. Biotech. Res., Vol 2 (1), 2012.
  3. Gomashe AV, Gulhane PA and Bezalwar PM, “Isolation and Screening of Cellulose Degrading Microbes from Nagpur Region Soil”, Int. J. of Life Sciences, Vol. 1(4), 2013.

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FAQs

What various products do bacteria produce?

Food additives, alcoholic and non-alcoholic beverages, biofuels, metabolites, biofertilizers, enzymes, bioactive molecules, vaccines, antibiotics, etc.

At what stage of growth do microorganisms produce products?

In the stationary phase, microorganisms produce products, which are classified as primary and secondary metabolites.

Examples of industrially important microorganisms?

Saccharomyces cerevisiae, Escherichia coli,Aspergillus niger,Clostridium butyclicum, Xanthomonas campestri, Deinococcus radiorans, etc.