STABILIZING EFFECT OF PENICILLIN-G SULFOXIDE, A COMPETITIVE INHIBITOR OF PENICILLIN-G ACYLASE - ITS PRACTICAL APPLICATIONS

被引:20
作者
ALVARO, G [1 ]
FERNANDEZLAFUENTE, R [1 ]
BLANCO, RM [1 ]
GUISAN, JM [1 ]
机构
[1] CSLC,INST CATALISIS,SERRANO 119,E-28006 MADRID,SPAIN
关键词
PENICILLIN-G ACYLASES; COMPETITIVE INHIBITORS OF; STABILIZING EFFECT OF COMPETITIVE INHIBITORS; IMMOBILIZATION-STABILIZATION OF PENICILLIN-G ACYLASES; ENZYME-SUPPORT MULTIPOINT COVALENT ATTACHMENT;
D O I
10.1016/0141-0229(91)90130-3
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
We have found that penicillin G sulfoxide (pen G SO) behaves as a general stabilizing agent of two bacterial penicillin G acylases (PGAs) (from E. coli and from K. citrophila), and this role is related to a strong inhibitory effect on the enzymes. The stabilizing effect has been observed during two different inactivation processes: (i) thermal inactivation of soluble enzymes at alkaline pH, and (ii) inactivation of immobilized enzymes as a consequence of covalent multiinteraction with highly activated agarose aldehyde gels. At the same time, pen G SO behaves as a strong competitive inhibitor of these two enzymes. The inhibition constant is more than 10-fold lower than the one corresponding to another smaller competitive inhibitor, phenylacetic acid (PAA), the structure of which is exactly the acyl donor moiety corresponding to pen G SO. In turn, PAA hardly exerts any stabilizing effect on PGAs. The stabilizing effect of pen G SO allowed the preparation of derivatives of these PGAs preserving full catalytic activity in spite of being 1,400- and 650-fold more stable than the corresponding soluble or one-point attached immobilized enzymes.
引用
收藏
页码:210 / 214
页数:5
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