Effects of Various Inhibitors on ?-galactosidase Purified from the Thermoacidophilic Alicyclobacillus acidocaldarius Subsp Rittmannii Isolated from Antarctica
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Tarih
2011
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Korean Soc Biotechnology & Bioengineering
Erişim Hakkı
info:eu-repo/semantics/closedAccess
Özet
beta-Galactosidase purified from the thermoacidophilic Alicyclobacillus acidocaldarius subsp. rittmannii isolated from Antarctica is a member of the GH42 family. The enzyme was not effected by various concentrations of its reaction product glucose, but was greatly inhibited by the other reaction product galactose using both substrates, ONPG and lactose. Linewever-Burk plot analysis derived from both ONPG and lactose hydrolysis results showed that galactose is a mixed-type inhibitor of the purified beta-galactosidase. The enzyme was slightly activated by Mg(2+) (13% at 20 mM), while inhibited at higher concentrations of Ca(+2) (33% at 10 mM), Zn(+2) (86% at 8 mM) and Cu(+2) (87% at 4 mM). The enzyme activity was not significantly altered by the metal ion chelators EDTA and 1,10-phenanthroline up to 20 mM, indicating that this enzyme is not a metalloenzyme. 2-Mercaptoethanol and DTT were found to enhance beta-galactosidase activity, while p-chloromercuribenzoic acid (PCMB) completely inhibited enzymatic activity (97% at 1 mM; 99.7% at 2 mM), indicating at least one essential Cys residue modified by the reagents in the active site of beta-galactosidase. Iodoacetamide and N-ethylmaleimide had little effect on the beta-galactosidase. Phenylmethylsulfonyl fluoride (PMSF) inhibited the enzyme strongly (19.8% at 1 mM; 71.9% at 10 mM), also showing the participation of serine for enzyme activity.
Açıklama
Anahtar Kelimeler
Beta-Galactosidase, Metal Ion Chelators, Inhibition, Galactose, Pmcb And Pmsf
Kaynak
Biotechnology and Bioprocess Engineering
WoS Q Değeri
Q3
Scopus Q Değeri
Q2
Cilt
16
Sayı
1
