Modified bicinchoninic acid assay for accurate determination of variable length reducing sugars in carbohydrates

Mohd Hasnain Md Hussain, . and Zainab Ngaini, . and Chong N. F. M., . Modified bicinchoninic acid assay for accurate determination of variable length reducing sugars in carbohydrates. pp. 2614-2619. ISSN 2231-7546

The accurate determination of reducing ends of malto-oligosaccharides is essential for calculating the enzyme activities of starch debranching enzymes. The suitability of the 3 5-Dinitrosalicylic acid (DNS) method the Dygert method and the Bicinchoninic acid (BCA) method for accurate determination of reducing ends from malto-oligosaccharides of different chain lengths is compared. The results showed that BCA assay was much more accurate than the other assays. The results for the BCA assay showed that different malto-oligosaccharides gave observed (measured) values that were significantly similar to the expected (predetermined) values. In contrast the DNS and Dygert assays underestimated the amount of reducing sugar present for glucose. Furthermore both DNS and Dygert methods showed increasing degree of overestimation of the amount of reducing sugar present with the increasing length of the malto-oligosaccharide sugar chains. The BCA assay can suitably quantify reducing sugars even in mixtures of oligosaccharides with different chain lengths. Thus enzyme activities can be measured without bias towards higher values for enzymes that preferentially cleave the longer chain lengths.

Full text available from: Official URL: http://www.ifrj.upm.edu.my/25%20(06)%202018/(50).p...

Abstract

The accurate determination of reducing ends of malto-oligosaccharides is essential for calculating the enzyme activities of starch debranching enzymes. The suitability of the 3 5-Dinitrosalicylic acid (DNS) method the Dygert method and the Bicinchoninic acid (BCA) method for accurate determination of reducing ends from malto-oligosaccharides of different chain lengths is compared. The results showed that BCA assay was much more accurate than the other assays. The results for the BCA assay showed that different malto-oligosaccharides gave observed (measured) values that were significantly similar to the expected (predetermined) values. In contrast the DNS and Dygert assays underestimated the amount of reducing sugar present for glucose. Furthermore both DNS and Dygert methods showed increasing degree of overestimation of the amount of reducing sugar present with the increasing length of the malto-oligosaccharide sugar chains. The BCA assay can suitably quantify reducing sugars even in mixtures of oligosaccharides with different chain lengths. Thus enzyme activities can be measured without bias towards higher values for enzymes that preferentially cleave the longer chain lengths.

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Item Type:	Article
AGROVOC Term:	Carbohydrates
AGROVOC Term:	Reducing sugars
AGROVOC Term:	Oligosaccharides
AGROVOC Term:	Enzyme activity
AGROVOC Term:	Starch
AGROVOC Term:	Glucose
AGROVOC Term:	Maltose
AGROVOC Term:	Isoamylase
AGROVOC Term:	Amylopectin
AGROVOC Term:	Oxidation
Depositing User:	Ms. Suzila Mohamad Kasim
Last Modified:	24 Apr 2025 06:29
URI:	http://webagris.upm.edu.my/id/eprint/24961

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