 |
|
Title
|
|
Expression, purification and crystallization of penicillin G acylase from Escherichia coli ATCC 11105.
|
|
|
Authors
|
|
P.D.Hunt,
S.P.Tolley,
R.J.Ward,
C.P.Hill,
G.G.Dodson.
|
|
|
Ref.
|
|
Protein Eng, 1990,
3,
635-639.
|
|
|
PubMed id
|
|
|
|
|
|
Abstract
|
|
|
The penicillin acylase gene (pac) from Escherichia coli ATCC 11105 was cloned
into pUC 9 and the resulting vector (pUPA-9), when transformed into E. coli
strain 5K, allowed the constitutive overproduction of mature penicillin acylase
when grown at 28 degrees C. The enzyme was purified from the periplasmic
fraction of E. coli pUPA-9 by hydrophobic interaction chromatography and anion
exchange. Crystals of penicillin acylase were grown in batch using polyethylene
glycol 8000 as a precipitant. The crystals (space group P1) diffracted to beyond
2.3 A.
|
|
|
|