Screening of PHB (Poly- β- hydroxybutyrate) Producing Bacteria

               

Aim:

          To Screen the PHB producing Bactria.

Introduction:

          Bacterial Cytoplasm contains a large number of inclusions called cytoplasmic inclusions for example volutin granules, metachromatic granules, phosphate granules,  Poly- β- hydroxybutyrate granules etc. The PHB is active microbial polyester and has been observed in many bacteria including rhizobia. It is an internal reserve food due to which bacterial cells survive to extended period of starvation in soil.

Materials Required:

Bacterial culture (Alcaligenes sp., Rhizopbia and Bacillus Sp.,)

0.3 % Sudan Black B solution (in ethylene glycol)

Safranin

Microscopic slide

Cover slip

Microscope

Procedure:

        1.    Prepared thin bacterial  smear on a slide surface, and thoroughly air dried

2.    Stained the smear with sudan black b solution for 5-15 min.

3.    Drained and dry out the slide and washed the slide with distilled water.

4.    Counter stained the slide with safranin for 10 -20 seconds.

5.    Then rinsed the slide in distilled water and blot dried with tissue paper.

6.    Observed the slide under microscope.  

1. 

Result:

         

          Blue droplets of Poly- β- hydroxybutyrate were observed, while cytoplasm appeared pink in oil immersion microscopy.