Enhancement of L-Asparaginase Production by Isolated
Bacillus circulans (MTCC 8574) Using Response Surface
M. Hymavathi & T. Sathish & Ch. Subba Rao &
R. S. Prakasham
Received: 13 August 2008 /Accepted: 11 November 2008
# Humana Press 2008
Abstract L-asparaginase production was optimized using isolated Bacillus circulans (MTCC
8574) under solid-state fermentation (SSF) using locally available agricultural waste
materials. Among different agricultural materials (red gram husk, bengal gram husk, coconut,
and groundnut cake), red gram husk gave the maximum enzyme production. Awide range of
SSF parameters were optimized for maximize the production of L-asparaginase. Preliminary
studies revealed that incubation temperature, moisture content, inoculum level, glucose, and
L-asparagine play a vital role in enzyme yield. The interactive behavior of each of these
parameters along with their significance on enzyme yield was analyzed using fractional
factorial central composite design (FFCCD). The observed correlation coefficient (R2) was
0.9714. Only L-asparagine and incubation temperature, were significant in linear and
quadratic terms. L-asparaginase yield improved from 780 to 2,322 U/gds which is more than
300% using FFCCD as a means of optimizing conditions.
Keywords L-Asparaginase . Bacillus circulans .
Fractional factorial central composite design . Optimization .
Red gram husk . Solid-state fermentation
L-Asparaginase (EC.184.108.40.206; asparagine amidohydrolase) has been widely used for the
chemotherapy of acute lymphoblastic leukemia and is gaining commercial importance in
the pharmaceutical sector. This enzyme is produced by a wide range of organisms including
animals, microbes, plants, and in the serum of certain rodents but not in human beings .
L-asparaginase production pattern was studied in Escherichia coli, Erwinia cartovora,
Corynebacterium glutamicum, Cylindrocarpon obtusisporum, Pseudomonas stutzeri,
Appl Biochem Biotechnol
M. Hymavathi : T. Sathish : C.