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The Pharma Innovation Journal 2019; 8(4): 18-20
ISSN (E): 2277- 7695
ISSN (P): 2349-8242
NAAS Rating: 5.03
TPI 2019; 8(4): 18-20
© 2019 TPI
www.thepharmajournal.com
Received: 11-02-2019
Accepted: 13-03-2019
Dani Benchamin
PG & Research, Department of
Zoology, St. Stephen’s College,
Pathanapuram, University of
Kerala, India
Sreejai R
P.G & Research Department of
Zoology, St. Stephen’s College,
Pathanapuram, University of
Kerala, India
Jensy Roshan F
PG & Research Department of
Zoology, St. Stephen’s College,
Pathanapuram, University of
Kerala, India
Sujitha S
PG & Research Department of
Zoology, St. Stephen’s College,
Pathanapuram, University of
Kerala, India
Beena S Kurup
PG & Research Department of
Zoology, St. Stephen’s College,
Pathanapuram, University of
Kerala, India
Albert C
Department of Zoology &
Research Centre, Scott Christian
College (Autonomous) Nagercoil,
Tamil Nadu, India
Correspondence
Dani Benchamin
PG & Research, Department of
Zoology, St. Stephen’s College,
Pathanapuram, University of
Kerala, India
Morphological and molecular identification of L-
Asparaginase producing fungus from Annona muricata
Dani Benchamin, Sreejai R, Jensy Roshan F, Sujitha S, Beena S Kurup,
Albert C
Abstract
The identification of L-Asparaginase producing fungal organisms has gained considerable attention
because of its less allergic properties. The present investigation deals with the morphological and
molecular identification of L-Asparaginase producing fungi from the leaves of the medicinal plant A.
muricata. It was confirmed that the fungal isolate obtained from A. muricata has the potential to produce-
Asparaginase and the fungal species was identified as Aspergillus fumigatus.
Keywords: L-Asparaginase, Annona muricata, Aspergillus fumigatus, Czapek Dox medium
Introduction
The tropical plant species A. muricata is very well known for its medicinal properties. The
plant parts such as bark, leaves, seed and fruit have been used as a curative substance for a
wide variety of health issues
[1]
. The anticancer enzyme L-Asparaginase has been derived from
various microorganisms isolated from different parts of many medicinal plants
[2, 3]
.
Surprisingly A. muricata is also a rich source of L-Asparaginase producing fungus
[4].
The L-
Asparaginase is from a fungal source is more suitable than other microbial sources because of
its less allergic properties when it is used in the treatment of cancer
[5]
. Acute lymphoblastic
leukemia is the condition where the blood cells become cancerous and in this case, L-
Asparaginase is an amazing remedial measure for the treatment of acute lymphoblastic
leukemia. The detection of L-Asparaginase as an anticancer drug and its further investigation
began in 1953. Researchers found that the guinea pig serum has the capability to fight against
lymphomas in rat and mice. Later they discovered that the L-Asparaginase enzyme present in
guinea pig serum was responsible for the prevention of lymphomas
[6, 7]
. Microorganisms are
widely used for the production of L-Asparaginase because of their effortless optimization and
high yield by genetic modification. Nowadays fungal L-Asparaginase is preferred by many
researchers because of their reduced side effects. The reviews showed that the Genus
Aspergillus has the ability to produce L-Asparaginase
[8]
.
The identification of L-Asparaginase producing fungi up to molecular level is very essential in
anti-cancer research and it will be helpful for the enormous production of L-Asparaginase. The
present investigation deals with the morphological and molecular identification of fungus
isolated from the medicinal plant A. muricata.
Materials and Methods
Collection of plant sample
The leaf samples were collected from the medicinal plant A. muricata growing in the southern
Western Ghat regions of Kerala.
Isolation and screening for L-Asparaginase
Mortar and pestle was used to ground the samples to very fine paste and serial dilution was
used for the isolation of fungal organism
[9]
. The modified Czapek Dox agar medium with a
pH 6.2 and phenol red as indicator were employed in the screening for L-Asparaginase. The
formation of pink color around the colonies indicated the production of L-Asparaginase
[10]
.
Identification of fungal species
The morphological and microscopical characteristics of isolated fungi were identified by
following the method Lacto Phenol Cotton Blue Staining
[11]
. For the molecular identification,