Title: Modification of the standard enzymatic protocol to a cost-efficient format for mixed-linkage (1¿3, 1¿4)-¿-D-glucan measurement Authors
Submitted to: Journal of Agricultural and Food Chemistry
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: April 1, 2008
Publication Date: September 1, 2008
Repository URL: http://riley.nal.usda.gov/nal_web/digi/submission.html
Citation: Hu, G., Burton, C. 2008. Modification of the standard enzymatic protocol to a cost- efficient format for mixed-linkage (1¿3, 1¿4)-ß-D-glucan measurement. Cereal Chemistry 85: 648-653. Interpretive Summary: The manuscript titled "Modification of the standard enzymatic protocol to a cost-efficient format for mixed-linkage (1->3, 1->4)-B-D-glucan measurement" is to improve the current beta-glucan measurement method for high throughput format and low cost purpose. The current available methods in B-glucan assay either lack the measurement accuracy or are low throughput and costly. They are difficult to use given the accuracy and large sample size required in experiments such as QTL mapping. In this manuscript we report the modified procedures for the enzymatic assay of B-glucan in small grains. The modifications enable the enzyme-based measurement of grain B-glucan to be upgraded into a high throughput format. The modified procedure dramatically reduced the cost on each sample to 20% of the original commercial protocol. The statistical analysis proved that the modified protocols are comparable to the standard one in accuracy and in precision of the measurements. The optimized protocol will provide a very useful tool in B-glucan studies in both genetic and breeding programs where a large number of samples are involved.
Technical Abstract: Abstract The current enzymatic assay approach (AACC Method 32-23) for the measurement of mixed-linkage B-glucan in small grains was modified to a cost-efficient and high throughput format without compromising the accuracy of the results. Ten barley (Hordeum vulgare L.) genotypes used in the study represented a wide range of B-glucan content levels. The reduced reaction volume is used in the new protocol to adapt to a 96-well plate format. Key components lichenase and B-glucosidase were reduced to 25% and the cost per sample was reduced to 22% of that in the original protocol. The accuracy of the measurement from the modified protocol is comparable to the current standard enzymatic procedure. B-glucan measurement accuracy of the modified and original protocols was also compared using 21 oat (Avena sativa L.) samples. The results indicated that the new protocol consistently produced accurate measurements in both barley and oat.