Characterization of Acidic Protease from Aspergillus niger BCRC 32720

Abstract

An acid protease from the broth of a 24 h cultivated Aspergillus niger BCRC 32720 was purified to electrophoretical homogeneity by CM Sepharose FF and Sephacryl S-100 HR chromatographs. The specific activity, purification fold, and yield were 23.29 kU/mg, 2.5, and 24.2%, respectively. Molecular mass (M) and N-terminal amino acid sequence were 47.5 kDa and SKGSAVTT, whereas the pH and temperature optima were at 2.5 and 50 °C, respectively. It was stable at pH 2.0-4.0 or ≤40 °C and activated by Fe(2+) and cysteine, but partially inhibited by phenylmethanesulfonyl fluoride and tosyllysine chloromethyl ketone and highly inhibited by Ag(+), Sn(2+), Fe(3+), Sb(3+), and pepstatin A. It was considered to be an aspartic protease.

Partial Characterization of an Acidic Protease from Rhizopus stolonifer RN-11

“…2). Furthermore, ≥70% activity remained after 24 hours of incubation at room temperature at pH ranges of 2.0-4.0, in conformity with the results reported by Li [27], Jose [23], Eneyslaya [28], Yin [24], and Chen [25]. According to the studies conducted by Hsiao [29], Kumar [11], Li [33] and Xu [35], the optimal pH values of extracellular acid protease from Rhizopus oryzae, and Aspergillus spp.…”

“…The acidic protease was purified 41.8-fold and the recovery yielded 24.4% after Sephadex G-75 chromatography ( Table 1). Acidic proteases of varying molecular weights were previously isolated from bacteria, including an aspartate protease from Phycomyces blakesleeanu (35 kDa) [23], aspartate protease from Rhizopus oryzae (34 kDa [11] and 47.5 kDa [24] variants), protease A(37 kDa) and protease B (34 kDa) from Aspergillus niger [25], Aspartic protease Sapt1p (44 kDa), Sapt2p (49 kDa), Sapt3 (55 kDa proproitein and 48 kDa mature protein) [10], and two forms of acid protease (M1 150 kDa and M2 60 kDa) from Aspergillus oryzae [26]. Unlike previously reported data, the purified acidic protease in this study was a monomer with an MW of ~70 kDa according to SDS-PAGE analysis (Fig.…”

“…Some metal ions seemed to have various effects on the activity of acidic proteases from different fungi. [24]. Cu 2+ strongly inhibited the acidic protease from A. niger [3].…”

Kinetics Study of Extracellular Detergent Stable Alkaline Protease from Rhizopus oryzae

“…The active fractions from this peak was pooled and lyophilized for further use. Yin et al (2013) obtained purification fold of 6.6 with specific activity of 117.62 KU/mg using sephacryl S-200 HR column chromatography. Anandan et al (2007) achieved purified fractions having specific activity of 905.7 using sephadex G-100 column chromatography.…”

Onychomycosis - a kórokozók fajspektruma és terápia

“…A mintában kimutatott penész ezen kívül laboratóriumi kontaminációnak is tekinthetô. A penészgombák kórokozó szerepét igazolja azonban, hogy egyes fajok keratináz és proteáz enzimeket termelnek súlyos szerkezeti deffektust okozva a körmökön (15). A penészgombák közül egy német és egy iráni tanulmánnyal ellentétben a vizsgált magyar betegpopulációban az Aspergillus niger (97,14% kézkörömben, 54,8% lábkö-römben) volt a leggyakoribb kórokozó (16,17).…”