Secretory Expression, Purification, Characterization, and Application of an Aspergillus oryzae Prolyl Aminopeptidase in Bacillus subtilis.

cDNA coding a prolyl aminopeptidase (PAP) was cloned from Aspergillus oryzae and over expressed in Bacillus subtilis with a 6×His tag in N-terminus. The recombinant prolyl aminopeptidase was secreted to extracellular by adding 2 mM CaCl and 5% D-sorbitol in TB medium; the enzyme activity in fermented supernatant increased from 7.2 to 41.

Over-expression of a proline specific aminopeptidase from Aspergillus oryzae JN-412 and its application in collagen degradation.

A strain that exhibited intracellular proline-specific aminopeptidase (PAP) activity was isolated from soy sauce koji and identified as Aspergillus oryzae JN-412. The gene coding PAP was cloned and efficiently expressed in Escherichia coli BL21 in a biologically active form. The highest specific activity reached 52.

A thermo-stable lysine aminopeptidase from Pseudomonas aeruginosa: Isolation, purification, characterization, and sequence analysis.

Pseudomonas aeruginosa NJ-814, isolated from garden soil, produced an extracellular aminopeptidase that was purified using ammonium sulfate precipitation and ion exchange chromatography. The purity was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the Mr value of the enzyme was estimated to be 55 kDa. The purified enzyme shows maximum activity at pH 9.

Purification and characterization of a urethanase from Penicillium variabile.

Urethanase produced by Penicillium variabile was purified through ultrasonication, concentration by polyethylene glycol 20,000, and Superdex G-200 gel filtration chromatography. The molecular weight of urethanase was determined to be around 96 kDa by gel filtration. The purified enzyme showed a single band in SDS-PAGE with the molecular weight of ~13.

Isolation and characterization of urethanase from Penicillium variabile and its application to reduce ethyl carbamate contamination in Chinese rice wine.

A strain with urethanase activity was isolated from mouse gastrointestine. By combination of morphological characterization of the colony, hyphae, and spore and the sequence analysis of its rDNA ITS, the strain was determined as Penicillium variabile and named as P. variabile JN-A525.

Purification of a novel pepsin inhibitor from Coriolus versicolor and its biochemical properties.

A novel pepsin inhibitor was isolated from Coriolus versicolor. The purification was carried out by a 2-step ultrafiltration followed by DEAE-52 and Mono Q ion-exchange chromatography. SDS-PAGE and gel filtration chromatography analysis showed that the isolated inhibitor was a 22.