New idea about LacZ

u6064043 at ucsvc.ucs.unimelb.edu.au u6064043 at ucsvc.ucs.unimelb.edu.au
Tue Jun 9 20:54:14 EST 1992


Hi there,

I am working on a class of proteoglycan (>80% carbohydrate) which is rich in
galactose (mainly Beta-1-3-galactoside chains) and have been struggling with
the deglycosylation condition (so that I can purify the protein backbone and
clone it) for quite a while. The method I used was anhydrous HF hydrolysis
as it was used in many other cases to deglycosylate O-linked Gal-Hypro linkage.
However I am always wondering about enzymetic method (chemical deglycosylation
always is difficult to predict, e.g., the effect of acidic condition on 
polypeptide chains). Then I came across with this funny idea: we have been
using LacZ (Beta-galactosidase) for cloning for years. Can this bacterial
enzyme help me in hydrolyse galactosyl chains? I went through a few early
papers about LacZ, all they tried was to hydrolyse dissachrise containing
galactosyl unit. Can it also degrades polysacchride (e.g., 1-3-galactan)?
Thank you in advance.



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