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A simple and efficient diffusion technique for assay of endo β-1,4-xylanase

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Title A simple and efficient diffusion technique for assay of endo β-1,4-xylanase
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Creator Samanta AK
Senani S
Kolte AP
Sridhar M
Jayapal N
 
Subject Endo-β-1
4-xylanase
Diffusion technique
Congo red assay
 
Description Not Available
Endo-β-1, 4-xylanases is thought to be of great significance for several industries namely paper, pharmaceuticals, food, feed etc. in addition to better utilization of lignocellulosic biomass. The present investigation was aimed to develop an easy, simple and efficient assay technique for endo-β-1, 4-xylanases secreted by the aerobic fungi. Under the proposed protocol, 9 g/L xylan containing agar was prepared in 100 mM phosphate buffer at different pH (4.5, 5.5 and 6.5). The sterilized xylan agar was dispensed in 90 mm petri dishes. 100 µl of culture supernatant of 12 fungal isolates was added to the wells and left overnight at 31±10C. The petri dishes were observed for zone of clearance by naked eye and diameter was measured. Congo red solution (1 g/L) was applied over the petri dishes as per the established protocol and thereafter plates were flooded with 1M Sodium chloride solution for the appearance of zone of clearance. The diameter for zone of clearance by the proposed method and the established protocol was almost identical and ranged from 21 to 42 mm at different pH depending upon the activity of endo-β-1, 4-xylanases. Change of pH towards alkaline side enabled similar or marginal decrease of diameter for the zone of clearance in most of the fungal isolates. The specific activities of these fungal isolates varied from 1.85 to 11.47 IU/mg protein. The present investigation revealed that the proposed simple diffusion technique gave similar results as compared to the established Congo red assay for endo-β-1, 4-xylanases. Moreover, the present technique avoided the cumbersome steps of staining by Congo red and de-staining by sodium chloride.
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Date 2021-08-04T06:02:34Z
2021-08-04T06:02:34Z
2012-01-01
 
Type Journal
 
Identifier Samanta AK, Senani S, Kolte AP, Sridhar M and Jayapal N. 2011. A simple and efficient diffusion technique for assay of endo β-1,4-xylanase. Brazilian Journal of Microbiology, 42:1349-1353
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http://krishi.icar.gov.in/jspui/handle/123456789/52840
 
Language English
 
Relation Not Available;
 
Publisher Not Available