ISOLATION, PURIFICATION AND CHARACTERIZATION OF SERINE PROTEASE INHIBITOR PRODUCING ENDOPHYTIC FUNGUS Penicillium chrysogenum sp. AZX2011
AZHAR KAMAL
Department of Chemistry, Faculty of Science, Integral University, Lucknow, UP, India and Department of Biosciences, Faculty of Sciences, Integral University, Lucknow, UP, India
ABDUL RAHMAN KHAN
Department of Chemistry, Faculty of Science, Integral University, Lucknow, UP, India
SYED SAYEED AHMAD
Department of Bioengineering, Faculty of Engineering, Integral University, Lucknow, UP, India
SAHEEM AHMAD
Department of Biosciences, Faculty of Sciences, Integral University, Lucknow, UP, India
MOHD. HARIS SIDDIQUI
Department of Bioengineering, Faculty of Engineering, Integral University, Lucknow, UP, India
MOHD. SAJID KHAN *
Department of Biosciences, Faculty of Sciences, Integral University, Lucknow, UP, India
*Author to whom correspondence should be addressed.
Abstract
The endophytic fungus Penicillium chrysogenum sp. AZX2011 was isolated from the medicinal plant
Lawsonia inermis. The phylogenetic study of isolated fungus was carried out by 18s rRNA analysis.
The partial sequence of 18s rRNA was submitted to the Gen bank database which allotted accession
number KU253718.1. This fungal species produces a strong serine protease inhibitor (SPI) which
showed inhibition against trypsin as a model enzyme. Purity of purified SPI was confirmed by SDSPAGE
(Sodium Dodecyl Sulphate-Polyacrylamide gel electrophoresis) and MALDI-TOF (Matrixassisted
laser ionization- time-of-flight mass spectrometer) with molecular weight 18.5 kDa. Serine
protease inhibitor (SPI) was found to be stable from pH 4.0 to pH 10.0 and temperature 25°C to
60°C. However, this trypsin inhibitor did not show any anti-protease activity against other proteases
like papain, metallo-proteases and pepsin.
Keywords: Trypsin;, serine protease inhibitor, matrix-assisted laser ionization- time-of-flight mass spectrometer, N-α-Benzoyl-DA-argenine4-nitroanilide hydrochloride