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Asian Pacific Journal of Tropical Biomedicine (2012)1-6

1

Contents lists available at ScienceDirect

Asian Pacific Journal of Tropical Biomedicine
journal homepage:www.elsevier.com/locate/apjtb

Document heading

Statistical optimization of glutaminase production from Lactobacillus
casei using response surface methodology
Sajitha Nagamony*, Lakshminarayanan Ramasamy1,Vasanthabharathi venkataraman2, S. Jayalakshmi Singaram3
CAS in Marine Biology, Faculty of Marine Sciences, Annamalai University, Parangipettai, 608 502, Tamilnadu, India.

ARTICLE INFO

ABSTRACT

Article history:
Received 15 April 2011
Received in revised form 27 April
Accepted 28 June 2011
Available online 28 June 2011

Objective: To isolate and optimize the glutaminase producing bacteria usingResponse Surface
Methodology. Methods: In the present study, the water, sediment samples and mangrove detritus
were collected from the vellar estuary (Tamil Nadu, India) and the potential strains of glutamines
production were screened in minimal medium plates. The optimization of glutaminase enzyme
was done by response surface methodology. Result: Totally 25 strains were collected. Among themLactobacillus casei in was found to be the potential strain. Hence this potential strain was used for
further optimization using Response Surface Methodology, partial purification using ammonium
sulphate precipitation, characterization, stability, antimicrobial and antioxidant application. The
stability of glutaminase from Lactobacillus casei was studied and better stability was observed atpH 7 to 8. The thermal stability was observed at 30°C to 40°C. The SDS PAGE of glutaminase
enzyme shows two different molecular weights. The molecular weight of partially purified
glutaminase enzyme was 23kDa and 34kDa. The glutaminase enzyme shows moderate antibacterial
activity. The glutaminase enzyme showed antioxidant activity. Conclusion: Thus the present study
revealed that thisLactobacillus casei strain is highly suitable for industrial applications especially
in medicinal and food industries.

2011

Keywords:
Isolation
Screening
Identification
Optimization using RSM
Partial purification
Characterization
Antibacterial and Antioxidant.

1. Introduction
Enzymes are essential to life because they speed up
metabolic reactions to a very great extent, but do notundergo any change in themselves. Enzymes are chiral
catalysts, producing mostly or only one of the possible
stereo isomeric products. A chiral molecule is a type of
molecule that lacks an internal plane of symmetry and has a
non-super imposable mirror image. Examples can be found
for many chiral compounds comprising alcohols and diols,
amines, amides, 毩-amino acids, 毬-amino acids, aminoalcohols, carboxylic acids and esters. The fermentation of
wine, leavening of bread, curdling of milk into cheese, and
brewing of beer are few examples of enzymatic reactions.
T he uses of enzymes in medical field include killing
*Corresponding author: N.Sajitha *
CAS in Marine Biology, Faculty of Marine Sciences, Annamalai University, Parangipettai,
608 502, Tamilnadu-, India.
Email:sajimicro@gmail.com

disease-causing microorganisms, promoting wound healing
and diagnosing certain diseases. In the industry they are
used for degrading oil spills and wastes into harmless
compounds, in cleaning stains, and in fermentation
processes to make alcoholic beverages, acids, chemicals
etc.
L-Glutaminase (L-glutamine amidohydrolase EC 3.5.1.2)
catalyzes the hydrolysis of L-glutamine toL-glutamic
acid and ammonia . The use of enzyme as drug is a crucial
facet of today’s pharmaceutical industry. L-glutaminase
a potent anti-leukemic agent[1].The action of glutaminase
plays a major role in the cellular nitrogen metabolism of
both, prokaryotes and eukaryotes[2]. L-glutaminase can be
derived from plant as well as animal sources but microbial
enzymes are generally meeting the...
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