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A Note on the Estimation of Microbial Glycosidase Activities by Dinitrosalicylic Acid Reagent

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Title A Note on the Estimation of Microbial Glycosidase Activities
by Dinitrosalicylic Acid Reagent
 
Creator Sengupta, S
Jana, M L
Sengupta, D
Navneet, A K
 
Subject Drug Development/Diagnostics & Biotechnology
 
Description In the estimation of glycosidase activity by
dinitrosalicylic acid (DNS) reagent, the stoichiometry of
DNS reduction was reported to increase proportionately
with the increase in the number of glycosidic linkages
present in oligosaccharides liberated by the enzyme. The
relationship between increases in DNS reduction and
increases in the number of glycosidic bonds was found to
be represented by a part of a rectangular hyperbola. The
increase was optimum with disaccharide and insigni®-
cant when the degree of polymerization (DP) was ³10.
The di�erence did not arise as a result of the DNSA
discriminating between mono- and oligosaccharide oxi-
dation. The relationship stemmed from the acidity of the
hydroxyl group adjacent to the reducing group, which
repressed DNS reduction. The acidity is likely to decrease
with an increase in oligosaccharide chain length. It is
suggested that DNS reduction is actually optimum and
uniform for all oligosaccharides of DP ³ 10 and that it is
minimum for monosaccharide. Thus the introduction of
recti®cation factors in the estimation of glycosidase
activities by the DNS method appears to be justi®ed. Smith and Wood 1991) compared to those given by the
arsenomolybdate (AM) reagent of Nelson (1944) and
Somogyi (1952). Ruttloff et al. (1970) indicated that the
stoichiometry of DNS reduction increased linearly with
the increase in chain length of the reducing substrate
(oligosaccharide). Maltohexaose had 3.5 times more
reducing activity than glucose. Breuil and Sadder (1985)
also indicated the chain-length-dependent reduction of
DNS. We report that DNS reductions by mono- and
oligosaccharide are not directly proportional to chain
length; instead, reduction increases via a complex rela-
tionship. The relationship arose out of the fact that DNS
reduction was inhibited by the C2 hydroxyl group ad-
jacent to the potential aldehyde group, inhibition being
maximum with monosaccharide and negligible when the
degree of polymerization (DP) of oligosaccharide was 10
or more. A theoretical relationship similar to that ob-
served between increases in DNS reduction and the DP
values of oligosaccharides can also be deduced on the
basis of the above observation.
 
Publisher Springer Verlag
 
Date 2000
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://www.eprints.iicb.res.in/802/1/APPLIED_MICROBIOLOGY_AND_BIOTECHNOLOGY_53(_6)_732%2D735;2000[36].pdf
Sengupta, S and Jana, M L and Sengupta, D and Navneet, A K (2000) A Note on the Estimation of Microbial Glycosidase Activities by Dinitrosalicylic Acid Reagent. Applied Microbiology and Biotechnology, 53 (6). pp. 732-735.
 
Relation http://dx.doi.org/10.1007/s002530000327
http://www.eprints.iicb.res.in/802/