Chandrasekaran, M; Archana, Kishore; Sreeja, Chellappan; Soorej, Basheer M; Jasmin, C; Sarita,G Bhat; Elyas, K K(Springer, November 26, 2010)
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Abstract:
An alkaline protease from marine Engyodontium
album was characterized for its physicochemical
properties towards evaluation of its suitability for potential
industrial applications. Molecular mass of the enzyme by
matrix-assisted laser desorption ionization-mass spectrometry
(MALDI-MS) analysis was calculated as 28.6 kDa.
Isoelectric focusing yielded pI of 3–4. Enzyme inhibition by
phenylmethylsulfonyl fluoride (PMSF) and aprotinin
confirmed the serine protease nature of the enzyme.Km, Vmax,
and Kcat of the enzyme were 4.727 9 10-2 mg/ml,
394.68 U, and 4.2175 9 10-2 s-1, respectively. Enzyme
was noted to be active over a broad range of pH (6–12) and
temperature (15–65 C), withmaximumactivity at pH 11 and
60 C. CaCl2 (1 mM), starch (1%), and sucrose (1%) imparted
thermal stability at 65 C. Hg2?, Cu2?, Fe3?, Zn2?, Cd?, and
Al3? inhibited enzyme activity, while 1 mMCo2? enhanced
enzyme activity. Reducing agents enhanced enzyme activity
at lower concentrations. The enzyme showed considerable
storage stability, and retained its activity in the presence of
hydrocarbons, natural oils, surfactants, and most of the
organic solvents tested. Results indicate that the marine
protease holds potential for use in the detergent industry and
for varied applications.
Description:
J Ind Microbiol Biotechnol (2011) 38:743–752
DOI 10.1007/s10295-010-0914-3
Chandrasekaran, M; Jasmin, C; Sreeja, Chellappan; Sarita,G Bhat; Rajeev, Sukumaran K; Elyas, K K(Springer, January 10, 2010)
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Abstract:
An alkaline protease gene (Eap) was isolated
for the first time from a marine fungus, Engyodontium
album. Eap consists of an open reading frame of 1,161 bp
encoding a prepropeptide consisting of 387 amino acids
with a calculated molecular mass of 40.923 kDa. Homology
comparison of the deduced amino acid sequence of
Eap with other known proteins indicated that Eap encode
an extracellular protease that belongs to the subtilase
family of serine protease (Family S8). A comparative
homology model of the Engyodontium album protease
(EAP) was developed using the crystal structure of proteinase
K. The model revealed that EAP has broad substrate
specificity similar to Proteinase K with preference for
bulky hydrophobic residues at P1 and P4. Also, EAP is
suggested to have two disulfide bonds and more than two
Ca2? binding sites in its 3D structure; both of which are
assumed to contribute to the thermostable nature of the
protein.
Chandrasekaran, M; Sreeja, Chellappan; Jasmin, C; Soorej, Basheer M; Elyas, K K; Sarita,G Bhat(Elsevier, October 17, 2005)
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Abstract:
Engyodontium album isolated from marine sediment produced protease, which was active at pH 11. Process parameters influencing the
production of alkaline protease by marine E. album was optimized. Particle size of <425 mm, 60% initial moisture content and incubation at 25 8C
for 120 h were optimal for protease production under solid state fermentation (SSF) using wheat bran. The organism has two optimal pH (5 and 10)
for maximal enzyme production. Sucrose as carbon source, ammonium hydrogen carbonate as additional inorganic nitrogen source and amino acid
leucine enhanced enzyme production during SSF. The protease was purified and partially characterized. A 16-fold purified enzyme was obtained
after ammonium sulphate precipitation and ion-exchange chromatography. Molecular weight of the purified enzyme protein was recorded
approximately 38 kDa by SDS-PAGE. The enzyme showed maximum activity at pH 11 and 60 8C. Activity at high temperature and high alkaline
pH suggests suitability of the enzyme for its application in detergent industry