Figures & data
Fig. 1. Separation of N-Ac-(R)- and (S)-β-Phe acylase activities using a Phenyl Sepharose column.
Notes: The protein solution obtained by ammonium sulfate precipitation was loaded onto a Phenyl Sepharose column. The N-Ac-β-Phe acylase activity of the eluted fractions was measured and expressed as N-Ac-(R)-(closed circles) or (S)-β-Phe (open circles) acylase activity in the panel.
Fig. 2. SDS-PAGE analysis of the purified (A) N-Ac-(R)-β-Phe acylase and (B) N-Ac-(S)-β-Phe acylase.
Notes: In the left lanes (denoted with M), standard proteins were applied; their molecular masses are indicated. The purified acylases were loaded to the right lanes.
Table 1. Purification of N-Ac-(R)-β-Phe acylase and N-Ac-(S)-β-Phe acylase.
Fig. 3. Enantioselectivity of purified acylases.
Notes: N-Ac-(R,S)-β-Phe was incubated with either purified N-Ac-(R)- or (S)-β-Phe acylase. The reaction products were analyzed by HPLC. (A) N-Ac-(R,S)-β-Phe standard; (B) (R)-β-Phe standard; (C) (S)-β-Phe standard; (D) reaction products using N-Ac-(R)-β-Phe acylase as the enzyme source; (E) reaction products using N-Ac-(S)-β-Phe acylase as the enzyme source.
Fig. 4. Genetic organizations around the gene encoding (A) N-Ac-(R)-β-Phe acylase and (B) N-Ac-(S)-β-Phe acylase.
Notes: The extents and directions of the ORFs are indicated by thick arrows. Genes encoding N-Ac-β-Phe acylases are illustrated with solid arrows.
Table 2. Location and properties of sequenced genes and proteins predicted to be encoded by those genes.
Fig. 5. Optimum pH (A) and temperature (B) of N-Ac-(R)-β-Phe acylase and N-Ac-(S)-β-Phe acylase.
Notes: (A) N-Ac-(R)-β-Phe acylase (open symbols) and N-Ac-(S)-β-Phe acylase (closed symbols). The buffers used were 100 mM acetate (pH 3.5–5.5, ○●), 100 mM MES (pH 5.5–7.0, △▲), 100 mM Tris (pH 7.0–9.0, □■) and 100 mM borate (pH 9.0–11.0, ◊♦) (B) N-Ac-(R)-β-Phe acylase (open symbols) and N-Ac-(S)-β-Phe acylase (closed symbols).
