OPTIMALIZATION OF

CELLULOSE PRODUCTION
FROM BACTERIA ISOLATED
FROM SOIL
ANJASTARI
HENOKH
TORY RACHEL
LAURENT
FUADY MAQOFFA AHMAD
INTRODUCTION

The cellulolytic property of some bacterial genera such as Cellulomonas,

Cellvibrio, Pseudomonas sp [9]. Bacillus, and Micrococcus [7], was also reported.
Enzyme production is closely controlled in microorganisms and for improving its
productivity these controls can be ameliorated.
Cellulase yields appear to depend upon a complex relationship involving a
variety of factors like inoculums size, pH value, temperature, presence of
inducers, medium additives, aeration, growth time, and so forth.
Cellulase is produced by a large number of microorganisms. They are either cell
bound or extracellular. Although a large number of microorganisms can
degrade cellulose, only a few of them produce significant quantities of free
enzymes capable of completely hydrolysing crystalline cellulose.
continuous research for advances in speckled aspects for cellulose production
(such as cost, substrate specificity, and specific activity) is desired to achieve
improved technoeconomic feasibility.
EXPERIMENTAL

Cellulase-producing bacteria were isolated from soils dilute by pour

plate or spread plate using CMC agar medium. The plates were
incubated at 45, 50, and 55C for 24 hours. To indicate the cellulose
activity of the organisms, diameter of the clear zone around colonies
on CMC agar was measured. This clear zone is to determine the
cellulose activity of the selected bacterial isolate in medium. The
cellulose activity of the selected bacterial isolate in liquid medium.
Enzyme Production Medium. Production medium contained (g/L)
glucose 0.5 gm, peptone 0.75 gm, FeSO4 0.01 gm, KH2PO4 0.5 gm,and
MgSO4 0.5 gm.Ten millilitres of medium were taken in a 100 mL conical
flask. The flasks were sterilized in autoclave at 121C for 15min, and after
cooling, the flask was inoculated with overnight grown bacterial
culture. The inoculated medium was incubated at 37C inshaker
incubator for 24h. At the end of the fermentation period,the culture
medium was centrifuged at 5000rpm for 15min to obtain the crude
extract,which served as enzyme source
Process Optimization for Maximum Cellulase Production
pH
Flasks with broth containing the optimum concentration of substrate
and carbon source are taken and the pH of the broth is adjusted to
7.0, 8.0, 9.0, 10.0, and 11.0 in different flasks using 1 N HCl and 1 N
NaOH and sterilized.
Temperature
Production medium at pH 7 was inoculated with overnight grown
selected bacterial strain. The broth was incubated at different
temperatures from 35, 40, 45,50,55,and 60 C for 24h. At the endof
incubation period, the cell-free culture filtrate is obtained and used as
enzyme source.
Carbon Sources
The effect of various carbon sources such as starch, glucose, maltose,
lactose, and fructose at the concentration of 1 to 5% was examined in
the production medium.
RESULT AND DISCUSSION
Effect of pH
 Effect of Temperature
 Effect of Carbon Source
 Effect of Different Concentration of Carbon Source
 Effect of Nitrogen Source
 Effect of Agro-Based Waste Material
CONCLUSION

From the experiment was already performed, pseudomonas

fulorescens among E.Coli, bacillus subtilis, and serratia marscens. The
optimum temperature and pH were determined as 40oC and 9-11
pH and best carbon and nitrogen sources were glucose and
ammonium sulphate. So we can make a maximum celulase
production with this ideal formulation of media composition. After
optimization, the mass production was carried in 1 litre of optimized
media at 40oC for 48 hours at a pH of 10 on a rotary shaker at 110
rpm. bacteria have high growth rate but the couldnt widely used in
cellulase production. Cellulase yield depends on some factors are
inoculum size, pH, temperature, presence of inducers, medium
additives, aeration, growth time, and so forth. The high activity ad
stability celulase enzymes can use in various industrial and
biotechnology application.