Technical note: detection and quantification of supplemental fungal β-glucanase activity in animal feed

Walsh, Gary; Murphy, Richard; Killeen, Gerry F.; Headon, Denis R.; Power, Ronan F.

doi:10.2527/1995.7341074x

Cited by 51 publications

(27 citation statements)

References 6 publications

(6 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Ecient precipitation of the enzyme with ammonium sulfate or ethanol facilitates detection of very small amounts of the enzyme. Walsh et al (1995) have described a method for quanti®cation of b-glucanase activity based on the diameter of hydrolyzed zones in agar gels containing substrate. We believe that this approach can also be (1) and without (4) lichenan (0.5%) after 4 weeks of incubation at 25°C under normal light intensity.…”

Section: Discussionmentioning

confidence: 99%

The use of a thermostable β-glucanase gene from Clostridium thermocellum as a reporter gene in plants

et al. 1998

View full text Add to dashboard Cite

In order to take advantage of the high thermostability of its product, beta-1,3;1,4-glucanase (lichenase), we used a modified version of the licB gene from Clostridium thermocellum as a reporter gene for the analysis of gene expression in transformed plants. The coding region of the licB gene was truncated at both ends. The truncated enzyme retained its activity and thermostability. The modified gene (m-licB), with and without a plant leader peptide-encoding sequence, was expressed in tobacco plants under control of either the Agrobacterium octopine TR-DNA 2' gene promoter or the promoter of the gene for the small subunit of ribulose-1,5-bisphosphate carboxylase. Expression of licB can be measured quantitatively and accurately, the assay is sensitive and simple enough to be used for analysis of various gene fusion systems or for screening of transformants. The enzyme is very stable and remains active in tissue extracts even after storage for 1 year and survives many thawing-freezing cycles. The lichenase-encoding gene was expressed at high levels in transformed tobacco plants without any apparent detrimental effects on vegetative growth or flowering.

show abstract

Section: Discussionmentioning

confidence: 99%

The use of a thermostable β-glucanase gene from Clostridium thermocellum as a reporter gene in plants

et al. 1998

View full text Add to dashboard Cite

show abstract

“…Diffusion of the enzyme and subsequent degradation of the substrate causes clearing of zones that can be detected using staining procedures. Walsh et al (1995) used this technique to measure β-glucanase in feed. We have recently modified this technique to measure a range of other exogenous enzyme activities (Colombatto, Furtado, and Beauchemin, unpublished data), and are working on improving the quantitative ability of the technique.…”

Section: Biochemical Characterizationmentioning

confidence: 99%

A rationale for the development of feed enzyme products for ruminants

Beauchemin¹,

Colombatto²,

Morgavi³

2004

Can. J. Anim. Sci.

View full text Add to dashboard Cite

. 2004. A rationale for the development of feed enzyme products for ruminants. Can. J. Anim. Sci. 84: [23][24][25][26][27][28][29][30][31][32][33][34][35][36]. The use of exogenous cell wall degrading enzymes is an emerging technology that shows potential in terms of improving feed utilization by ruminants. This review discusses current information related to enzyme product formulation for ruminants, and addresses the conditions necessary to ensure effective and consistent in vivo results of providing feed enzymes to ruminants. Research has demonstrated that, in some cases, adding fibrolytic enzymes to dairy cow and feedlot cattle diets improves cell wall digestion and, consequently, weight gain or milk production are enhanced. However, considerable research is required to develop more effective enzyme products and to ensure consistency of responses in vivo. There is a need to identify the key enzyme activities involved in the positive responses observed in vivo and these enzyme activities should be assessed using a temperature and pH representative of the conditions in the rumen. However, to date, it has not been possible to accurately evaluate exogenous enzymes based only on their biochemical characterization because the model substrates used do not represent the complexity of plant cell wall material. In vitro techniques using feed substrates, buffer and ruminal fluid can be used more reliably as a bioassay to predict in vivo response to exogenous enzymes, however, other factors, including under or over-supplementation of enzyme activity, method of providing the enzyme product to the animal, composition of the diet, and the target animals must also be considered. Cet article passe en revue l'information existante sur la formulation des mélanges enzymatiques pour les ruminants et indique les conditions nécessaires pour obtenir des résultats in vivo utiles et conséquents lorsqu'on administre ces préparations aux animaux. Les recherches prouvent que, dans certains cas, l'addition d'enzymes fibrolytiques à la ration des vaches laitières et des bovins d'engrais entraîne une meilleure digestion de la paroi cellulosique, donc un gain de poids ou la production accrue de lait. Néanmoins, il reste encore beaucoup à faire avant qu'on mette au point de meilleurs produits et parvienne à des réactions in vivo uniformes. Ainsi, il faut identifier les principales activités enzymatiques à l'origine des réactions positives observées in vivo puis évaluer l'activité des enzymes à une température et à un pH représentatifs de ceux du rumen. Jusqu'à présent, on n'a pu évaluer avec précision les enzymes exogènes uniquement à partir de leurs propriétés biochimiques, les substrats employés dans le modèle ne reproduisant pas assez bien la complexité de la paroi cellulosique. Les techniques in vivo qui recourent aux aliments du bétail comme substrat, à des solutions tampons et aux fluides du rumen pourraient s'avérer biologiquement plus fiables pour prédire la réaction in vivo aux enzymes exogènes, mais on doit aussi prendre en compte d...

show abstract

“…Among them, endo-1,3-1,4-b-glucanases and endo-1,3(4)-b-glucanases have been widely applied as industrial enzymes in the brewing process to reduce the viscosity and filtration time of mash and in animal feed to improve the digestibility of b-glucan [2,7,38]. A number of bacterial glucanases have been reported from Streptomyces [33], Fibrobacter succinogenes [36], Bacillus [30,37], and Paenibacillus [10,39].…”

mentioning

confidence: 99%

High-level expression of a novel Penicillium endo-1,3(4)-β-d-glucanase with high specific activity in Pichia pastoris

Chen

Meng

Shi

et al. 2012

Journal of Industrial Microbiology and Biotechnology

View full text Add to dashboard Cite

A novel endo-1,3(4)-β-D-glucanase gene (bgl16C1) from Penicillium pinophilum C1 was cloned and sequenced. The 945-bp full-length gene encoded a 315-residue polypeptide consisting of a putative signal peptide of 18 residues and a catalytic domain belonging to glycosyl hydrolase family 16. The deduced amino acid sequence showed the highest identity (82%) with the putative endo-1,3(4)-β-glucanase from Talaromyces stipitatus ATCC 10500 and 60% identity with the characterized β-1,3(4)-glucanase from Paecilomyces sp. FLH30. The gene was successfully overexpressed in Pichia pastoris. Recombinant Bgl16C1 constituted 95% of total secreted proteins (2.61 g l⁻¹) with activity of 28,721 U ml⁻¹ in a 15-l fermentor. The purified recombinant Bgl16C1 had higher specific activity toward barley β-glucan (12,622 U mg⁻¹) than all known glucanases and also showed activity against lichenan and laminarin. The enzyme was optimally active at pH 5.0 and 55°C and exhibited good stability over a broad acid and alkaline pH range (>85% activity at pH 3.0-7.0 and even 30% at pH 11.0). All these favorable enzymatic properties make it attractive for potential applications in various industries.

show abstract

Technical note: detection and quantification of supplemental fungal β-glucanase activity in animal feed

Cited by 51 publications

References 6 publications

The use of a thermostable β-glucanase gene from Clostridium thermocellum as a reporter gene in plants

The use of a thermostable β-glucanase gene from Clostridium thermocellum as a reporter gene in plants

A rationale for the development of feed enzyme products for ruminants

High-level expression of a novel Penicillium endo-1,3(4)-β-d-glucanase with high specific activity in Pichia pastoris

Contact Info

Product

Resources

About