Research Article Improvement in Saccharification Yield of Mixed Rumen Enzymes by Identification of Recalcitrant Cell Wall Constituents Using Enzyme Fingerprinting Ajay Badhan, 1 Yu-Xi Wang, 1 Robert Gruninger, 1 Donald Patton, 2 Justin Powlowski, 2 Adrian Tsang, 2 and Tim A. McAllister 1 1 Agriculture and Agri-Food Canada, Lethbridge Research Centre, Lethbridge, AB, Canada T1H 4P4 2 Centre for Structural and Functional Genomics, Concordia University, Montreal, QC, Canada H4B 1R6 Correspondence should be addressed to Tim A. McAllister; tim.mcallister@agr.gc.ca Received 19 November 2014; Revised 15 January 2015; Accepted 16 January 2015 Academic Editor: Tean-Hock Tang Copyright © 2015 Ajay Badhan et al. Tis is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Identifcation of recalcitrant factors that limit digestion of forages and the development of enzymatic approaches that improve hydrolysis could play a key role in improving the efciency of meat and milk production in ruminants. Enzyme fngerprinting of barley silage fed to heifers and total tract indigestible fbre residue (TIFR) collected from feces was used to identify cell wall components resistant to total tract digestion. Enzyme fngerprinting results identifed acetyl xylan esterases as key to the enhanced ruminal digestion. FTIR analysis also suggested cross-link cell wall polymers as principal components of indigested fber residues in feces. Based on structural information from enzymatic fngerprinting and FTIR, enzyme pretreatment to enhance glucose yield from barley straw and alfalfa hay upon exposure to mixed rumen-enzymes was developed. Prehydrolysis efects of recombinant fungal fbrolytic hydrolases were analyzed using microassay in combination with statistical experimental design. Recombinant hemicellulases and auxiliary enzymes initiated degradation of plant structural polysaccharides upon application and improved the in vitro saccharifcation of alfalfa and barley straw by mixed rumen enzymes. Te validation results showed that microassay in combination with statistical experimental design can be successfully used to predict efective enzyme pretreatments that can enhance plant cell wall digestion by mixed rumen enzymes. 1. Introduction Rising grain prices heightened concerns over the use of food as feed for livestock production and the negative impacts of annual crops on carbon sequestration and biodiversity has prompted research into fnding ways to increase the use of fbrous forage in ruminant diets. Plant cell walls can consti- tute a primary source of nutritional energy for ruminants. However for many types of forage, less than 50% of the cell wall fraction is digested and utilized by the ruminant host [1]. Substantial benefts would be realized if a greater percentage of this potential energy was made available for fermentation in the rumen through an increase in the digestibility of the cell wall fraction. Fiber digestion in ruminants occurs primarily in the rumen and cecum. Generally, the amount of fber digested in the lower tract is relatively small, with the rumen being the primary site of digestion. Te ruminal microbial population secrets diverse hydrolases to degrade and ferment structural carbohydrates in plant cell walls. Te physical and chemical nature of forages can present a barrier to their complete digestion in the rumen [2]. Terefore, prior knowledge about the structural aspects of cell wall polymers that limit digestion is critical to identifying efcient enzymatic pretreatments. In this study enzyme fngerprinting was used in combina- tion with Flourier infrared spectroscopy (FTIR) to identify recalcitrant factors that limit fber digestion by mixed rumen enzymes. Hindawi Publishing Corporation BioMed Research International Volume 2015, Article ID 562952, 13 pages http://dx.doi.org/10.1155/2015/562952