Mohanasundaram S et al., (2017) Int. J. Res. Pharm. Sci., 8(3), 282-287
282 ©JK Welfare & Pharmascope Foundation | International Journal of Research in Pharmaceutical Sciences
Optimization of production of Extracellular proteolytic keratinase using
Streptomyces sp. isolated from poultry plant soil
Mohanasundaram S*
1
, Doss VA
2
, Prabhaharan CTA
1
, Joseph J
1
, Venugopal K
1
, Agilandeshwari P
3
, Sudansuhaa
C
1
, Balaganesh R
1
, Ramkumar R
1
, Rajaruban MDS
1
, Balaji B
1
1
Department of Biotechnology, Karpaga Vinayaga College of Engineering and Technology, Maduranthagam,
Tamil Nadu, India
2
Department of Biochemistry, PSG College of Arts and Science, Coimbatore, Tamil Nadu, India
3
Department of Biotechnology, Vivekananda College of Engineering for Women, Tiruchengode, Tamil Nadu, India
ABSTRACT
In this study, Proteolytic keratinase was observed from Streptomyces sp., isolated from poultry plant soil and puri-
fied by ammonium sulfate fractionation method followed by Gel filtration chromatography. The purified enzyme
was further characterized partially which covers the influence of pH, temperature, incubation time, nitrogen
sources and metal ions on the activity of proteolytic keratinase. From the results, it was observed that, proteolytic
keratinase isolated from the Streptomyces sp. enumerated from the poultry plant soil showed maximal activity at
pH 8.0 and at temperature 50oC within 55mins. It was also found that, at the minimum concentration of 2mM
BaCl.2H
2
O, CuSO
4
.5H
2
O and MgCl
2
, it showed highest keratinase activity.
Keywords: fermentation; Proteolytic keratinase; Streptomyces sp.; stability.
INTRODUCTION
Wool, bristle, horns, feathers, hoof, etc are some ex-
ample waste materials containing huge level of keratin
discharged from animal waste, leather and poultry
plants (Gousterova et al., 2005). Since these wastes are
rich sources of protein and amino acids, they can be
utilized as an essential component in animal feed
preparations. Few years before, these materials were
subjected into high temperature and subsequent mill-
ing for some other uses after degradation (Wang and
Parson, 1977). Keratin based wastes can be compe-
tently degraded by a few bacteria and fungi due to the
secretion of keratinolytic peptidases into the culture
medium (Onifade et al., 1998). Keratin-like materials
cannot be successfully degraded by the peptidases
currently included in detergents. Hence, enzyme with
high activity has to be finding out to meet the industri-
al requirements and for other applications. So far, wide
range of microorganisms with proteolytic keratinase
activity has been found, including some species of Ba-
cillus (Williams et al., 1990), Actinomycetes (Boeckle et
al., 1995) and fungi (Santos et al., 1996). Always, there
is a constant demand for thermophilic organisms be-
cause of their high thermal stability and resistance to-
wards denaturation by acidic and alkaline factors. The
most important feature of thermophilic organisms is
production of enzymes that catalyze biochemical reac-
tions at higher levels of temperatures and these en-
zymes are widely used in many industrial areas in
modern day of science. In recent years, many enzymes
from these thermophilic bacteria have been effectively
purified, characterized and used in industrial applica-
tions (Berna et al., 2015). The aim of this present study
is to isolate the Streptomyces species producing extra-
cellular proteolytic keratinase from the poultry plant
soil and subsequent purification and characterization
of the enzyme.
MATERIALS AND METHODS
Microorganism
Streptomyces sp. producing extracellular proteolytic
keratinase was isolated from the poultry plant soils
from in and around Thirukkalukundram, Kanchipuram
district, Tamil Nadu, South India (Dey et al., 1992 and
Hinge et al., 1989).
Chemicals
Solvent and other chemicals which were used in this
study were purchased from Himedia, Merck and s.d.
Fine-Chemicals, Mumbai, India.
Methods
Enzyme Production
Submerged fermentation was performed by inoculat-
ing pure culture of isolate into the production medium
www.ijrps.pharmascope.org
ISSN: 0975-7538
Research Article
* Corresponding Author
Email: sbmohan2007@gmail.com
Contact: +91-8870903888
Received on: 22-07-2017
Revised on: 29-07-2017
Accepted on: 06-08-2017