Publications

Authors:

Bombardi, Luca; Coltro, Annalaura; Fusco, Salvatore

Title:

Endo-1,4-β-D-xylanase Assay Using Azo-Xylan and Variants Thereof

Year:

2025

Type of item:

Articolo in Rivista

Tipologia ANVUR:

Articolo su rivista

Language:

Inglese

Referee:

No

Name of journal:

BIO-PROTOCOL

ISSN of journal:

2331-8325

N° Volume:

15

Number or Folder:

1369

Page numbers:

1-11

Keyword:

Azo-Xylan; Colorimetric assay; Dyed xylan; Enzymatic assay; Lignocellulosic biomass; Thermostability; Xylan degradation; Xylanases

Short description of contents:

: Xylan is the main component of hemicellulose and consists of a complex heteropolysaccharide with a heterogeneous structure. This framework, in addition to the crystalline structure of cellulosic fibers and the rigidity of lignin, makes lignocellulosic biomass (LCB) highly recalcitrant to degradation. Xylanases are glycoside hydrolases that cleave the β-1,4-glycoside linkages in the xylan backbone and have attracted increasing attention due to their potential uses in various industrial sectors such as pulp and paper, baking, pharmaceuticals, and lignocellulosic biorefining. For decades, the measurement of xylanase activity was based on reducing sugar quantification methods like DNS or Nelson/Somogyi assays, with numerous limitations in terms of specificity and interference from other enzymatic activities. A better alternative is the colorimetric Azo-Xylan assay, which specifically measures the endo-1,4-β-D-xylanase activity. In this study, the Azo-Xylan protocol was adapted from the company Megazyme to determine the enzymatic activity of thermostable xylanases produced by microbial consortia (i.e., microbiomes), aiming to determine biochemical features such as temperature and pH optima, thermostability, and shelf life. This modified approach offers a rapid, cost-effective, and highly specific method for the determination of xylanase activity in complex mixtures, helping the development of a xylanase-based method for the hydrolysis of hard-degrading substrates in bio-based industries. Key features • Direct enzyme assay for qualitative xylanase activity detection or quantitative measurement with a calibration curve. • Specific for determination of endo-1,4-β-D-xylanase activity, allowing to overcome interferences by enzymes with other activities.

Product ID:

145816

Handle IRIS:

11562/1162229

Last Modified:

May 16, 2025

Bibliographic citation:

Bombardi, Luca; Coltro, Annalaura; Fusco, Salvatore, Endo-1,4-β-D-xylanase Assay Using Azo-Xylan and Variants Thereof «BIO-PROTOCOL» , vol. 15 , n. 1369 , 2025 , pp. 1-11

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