Endo-P-galactosidase

For degradation of the carbohydrate moiety of 50 mg ARS2 sample, 10 or 25 U exo-P-galactosidase were added and the mixture was incubated at 37°C for 18 hr 10 ml of 100 mM acetate buffer (pH 4.2) containing 25 U endo-P-1,4-galactanase or 1 mg crude endo-P-1,6-galactanase [43] were added and the mixture was incubated at 45°C for 48 hr. Fifty unit exo-a-mannosidase or 0.5 U endo-P-galactosidase in 50 mM potassium phosphate buffer (pH 6.0) at 37 °C for 18 h was also used. 

M. Kitamikado, M. Ito and Y.-T. Li, Endo-p-galactosidase from Fiavobacterium keratolyticus. Methods Enzymol, 1982, 83, 619-625. 

P. Scudder, K.-I. Uemura, J. Dolby, M.N. Fukuda and T. Feizi, Isolation and characterization of an endo-P-galactosidase from Bacteroides fragilis. Biochem. J., 1983, 213, 485-494. 

S. Takasaki and A. Kobata, Purification and characterization of an endo-P-galactosidase produced by Diplococcus pneumoniae. J. Biol Chem., 1976, 251, 3603-3609. 

Hounsell, E.F., J. Feeney, P. Scudder, P.W. Tang, and T. Feizi. 1986. H-NMR studies at 500 MHz of a neutral disaccharide and sulphated di-, tetra-, hexa-, and larger oligosaccharides obtained by endo-P-galactosidase treatment of keratan sulphate. Eur. J. Biochem. 157 375-384. 

Incubation of SBP with CinnAE alone gave a single peak of absorbance at 310 nm corresponding to FA (0.91% of the alkali-extractable ferulic acid was released). No feruloylated material was released when SBP was incubated with a mixture of endo-(, 4)-p-D-galactanase and /3-D-galactosidase. There was no increase in FA release when CinnAE was supplemented with these two enzymes. 

A list of entrapped enzymes is given in Table 3.2. There are two endo-l,3-P-D-glucanases from marine mollusks Spisula sacchalinensis and Ch. albidus and a-D-galactosidase from marine bacterium Pseudoalteromonas sp. MM 701. They were selected for immobilization for the following reasons ... 

Hamaguchi, Y., Yoshitake, S., Ishikawa, E., Endo, Y., and Ohtaki, S. (1979) Improved procedure for the conjugation of rabbit IgG and Fab antibodies with p-D-galactosidase from Escherichia coli using N,N -o-phenylenedimaleimide./. Biochem. (Tokyo) 85,1289—1300. 

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