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Abstract

Production of cellulase is of great significance in present day biotechnology. Cellulose biodegradation by cellulases, produced by numerous microorganisms is very important in several agricultural and waste treatment processes. The development of microbial strains, media composition and process control has including submerged fermentation and solid state fermentation all contributed to achievements of high levels of cellulases for subsequent applications. One of these important applications is supplementing diets of farm animals with cellulases to improve feed utilization and animal performance by enhancing fiber degradation. Dairy cows feed forge treated with a cellulase enzyme preparations ate more feed and produced 5-25% more milk. This review provides an over view of the main variables to be considered for cellulase production from agricultural residues for animal feeding.
Endoglucanase
Exo-cellobiohydrolase I
Exo-cellobiohydrolase II
β-glucosidase
Cellobiose
Glucose
Cells
Transverse
Section of a log
Middle lamella
Primary wall
Secondary
Wall (S3)
Secondary
Wall (S2)
Secondary
Wall (S1)
Protein
Secondary wall
Middle lamella
Primary wall
Cellulose
Hemicellulose
Lignin
... Cellulose and xylanase are the two main evaluated fibredegrading enzymes (Zilio et al., 2019) in dairy cows. Inconsistent effects were found on nutrient utilisation (Kung et al., 2000;Yang et al., 2000;Elwakeel et al., 2007) and production performance (Murad and Azzaz, 2010;Dean et al., 2013;Kholif and Aziz, 2014). Recently, two meta-analyses (Arriola et al., 2017;Tirado-González et al., 2018) showed that supplementing diets with exogenous fibre-degrading enzymes has positive overall effects on lactation in dairy cows (Adesogan et al., 2019). ...
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We hypothesised that adding a combination of fibrolytic and amylolytic enzymes to the diet of early-lactation dairy cows would improve rumen enzyme activity and bacterial diversity, promote energy metabolism, and benefit milk production in cows. Twenty multiparous early-lactation (90 ± 5 d) Holstein cows with similar body conditions were randomly allocated to control (CON, n = 10) and experimental (EXP, n = 10) groups in a completely randomised single-factor design. The CON was fed only a basal total mixed ration diet, and the diet of the EXP was supplemented with a combination of fibrolytic and amylolytic enzymes at 70 g/cow/d (cellulase 3 500 CU/g, xylanase 2 000 XU/g, β-glucanase 17 500 GU/g, and amylase 37 000 AU/g). The experiment lasted 28 days, with 21 days for adaptation and 7 days for sampling. Enzyme addition increased the activity levels of α-amylase and xylanase, and the ammonia-N concentration (P 0.1%) or genus (richness > 1%) composition between the CON and EXP groups (P > 0.05). A tendency of difference was found between CON and EXP (R = 0.22, P = 0.098) in principal component analysis. Ten genera showed different abundances across the CON and EXP groups (linear discriminant analysis effect size, linear discriminant analysis > 2). EXP increased the ratio of albumin to globulin and the concentrations of total cholesterol and low-density lipoprotein cholesterol (P
... Also, using cellulase in combination with different enzymes removes nonnutritional substances (some lignocelluloses like lignin and cellulose, among others) in feed grains and green fodder by breaking them down. Hence, the enzyme benefits the animals (Asmare et al., 2014;Murad et al., 2010;Sharada et al., 2014). ...
Chapter
Endophytes are the microorganisms that inhabit plants without any detriment to the latter. These microbial communities are believed to be a long associated with plants and have evolved with them. Living inside the plants, these endophytes help the plants in dealing with various kinds of environmental or biological stresses, increasing the productivity and sustainability of the plant. Endophytes achieve this by producing certain bioactive compounds (secondary metabolites or enzymes), altering the physiology of the plant, acting as biocontrol agents for both pathogenic microbes and pests, etc. This chapter reviews a variety of fungal endophytes residing in crop plants and aiding crop improvement through various mechanisms.
... Since the use of cellulases in ruminants diets such as cattle is very common, stability against proteolytic digestion can be a significant benefit for exogenous cellulase enzymes 48,49 . An important application of cellulases enzymes is in the animal feed industry, which, alongside other enzymes, including proteases, improves feed utilization and animal performance 50,51 . ...
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Cellulases are hydrolytic enzymes with wide scientific and industrial applications. We described a novel cellulase, CelC307, from the thermophilic indigenous Cohnella sp. A01. The 3-D structure of the CelC307 was predicted by comparative modeling. Docking of CelC307 with specific inhibitors and molecular dynamic (MD) simulation revealed that these ligands bound in a non-competitive manner. The CelC307 protein was purified and characterized after recombinant expression in Escherichia coli (E. coli) BL21. Using CMC 1% as the substrate, the thermodynamic values were determined as Km 0.46 mM, kcat 104.30 × 10–3 (S−1), and kcat/Km 226.73 (M−1 S−1). The CelC307 was optimally active at 40 °C and pH 7.0. The culture condition was optimized for improved CelC307 expression using Plackett–Burman and Box–Behnken design as follows: temperature 20 °C, pH 7.5, and inoculation concentration with an OD600 = 1. The endoglucanase activity was positively modulated in the presence of Na+, Li+, Ca2+, 2-mercaptoethanol (2-ME), and glycerol. The thermodynamic parameters calculated for CelC307 confirmed its inherent thermostability. The characterized CelC307 may be a suitable candidate for various biotechnological applications.
... Major attention has been given to lignocellulose as a substrate for the production of cellulase enzymes. The component of lignocellulose materials is cellulose, hemicellulose and lignin [16]. Fibers can not be completely digested by animals and around 20-70% of cellulose is lost. ...
Article
In the present investigation, the production of amylase, cellulase, and protease activity by Bacillus subtilis ATCC 6051a was evaluated on different raw materials (soybean meal, peas, sorghum flour, corn) and combined feed (CF). The effect of various fermentation conditions (24-72 h) on enzymatic production through shake-flask culture (Erlenmeyer 100 mL) in optimum conditions (150 rpm, pH 7.0±0.2, 37◦C) was investigated. The inoculum strain presents 1.907 optical density (OD) 600 nm with a concentration of 1.6 x 109 CFU/mL. The screening showed a capacity of amylase, cellulase, and protease strain production. The maximum amylase level was obtained at 72 h when the strain was cultured in CF fermentation medium (13.19±0.15 U/mL), followed by corn (11.72±0.15 U/mL), peas (9.22±0.11 U/mL), soybean meal (7.29±0.19 U/mL) and sorghum (6.31±0.2 U/mL). For production of cellulase by DNS method, the activity was noticed in corn (4.39±0.04 U/mL), sorghum (4.05±0.02 U/mL), peas (2.94±0.06 U/mL), soybean meal (2.87±0.04 U/mL) and CF medium (1.41±0.02 U/mL). Maximum protease activity was observed after 24 h in CF (4.91±0.08 U/mL), with minimum enzyme production in the rest of fermentation media, both at 48 h and 72 h. In conclusion, qualitative analysis revealed that Bacillus subtilis ATCC 6051a could be successfully used for high scale production of amylase, cellulase and to less extend protease, particularly in fermented medium containing CF or corn, and may be considered a potential candidate for supplementation of animal feed.
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Chapter
Cellulose, an important structural component of the plant cell, is the most plentiful biological matter in the world, owing to its ubiquity in the entire plant kingdom. Cellulosic sugar is a renewable and sustainable source of fuels, biochemicals, and other value-added products. Among all type of cell wall carbohydrates, cellulose is not easily degraded through enzymatic catalysis. Fungal cellulases are a group of potential enzymes which are exploited for the bioconversion of lignocellulosic plant biomass for various commercial, environmental, and academic research purposes. This chapter presents the cellulase structure and its biodegradation by fungal cellulases, along with elaboration of various production methods. Overall, this chapter attempts to give a brief narration of the updated biotechnological state-of-the-art for cellulases.
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This book covers key areas of Microbiology and Biotechnology. The contributions by the authors include Elaeagnus conferta, Mimusops elengi, insecticidal activity, antimicrobial activity, fruit extract, mosquito borne diseases, prevention, Anaerobic bacteria, susceptibility testing, methodology, antimicrobial resistance, Bacteroides fragilis group, Clostridium spp., Phoenix dactylifera L., Barhee, in vitro, bacterial cell wall, initial pro callus, nitrogen, Bacterial media treatment, Broad bean seeds, germination in peeled form, microbiological indicators, bio-activated seed, Microbiological Safety, Poultry farms, Antibiotic resistance, Tetracycline, Erythromycin, Staphylococci, livestock industry, treatment and prevention of diseases, Antitumor activity, iodine complex, LiCl (I)- I2-a-dextrin- complexes, doxorubicin, quantumchemical approach, Lithium Halogenides, Bioorganic Ligands, Anti-cancer activity, a-dextrins, halogenides, antitumor drugs, Rumuola pond, Excavated Ponds, Antibiotic Resistance, Bacteria, Pefloxacin, Mimotopes, epitopes, Salmonella, E. coli, lipopolysaccharide, phagotopes, immunogenic peptides, synthetic peptides, phage display, Enterobacteria, Pleurotus, mushroom, inter simple sequence repeat (ISSR), internal transcribed spacer region, genetic diversity, Aspergillus niger, cellulase, microbial enzyme, purification and characterization, Biocatalyst, Basal metabolic rate, existence metabolism, endothermy, evolutionary and ecological scenario, erythrocytes in birds and mammals, aerobic supply of activity and sensory systems, minimal size, parental care etc. This book contains various materials suitable for students, researchers and academicians in the field of Microbiology and Biotechnology.
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Antibiotic growth promoters have been utilized for long time at subtherapeutic levels as feed supplements in monogastric animal rations. Because of their side-effects such as antibiotic resistance, reduction of beneficial bacteria in the gut, and dysbiosis, it is necessary to look for non-therapeutic alternatives. Probiotics play an important role as the key substitutes to antibacterial agents due to their many beneficial effects on the monogastric animal host. For instance, enhancement of the gut microbiota balance can contribute to improvement of feed utilization efficiency, nutrients absorption, growth rate, and economic profitability of livestock. Probiotics are defined as “live microorganisms that, when administered in adequate amounts, confer a health benefit on the host.” They are available in diverse forms for use as feed supplements. Their utilization as feed additives assists in good digestion of feed ingredients and hence, making the nutrients available for promoting growth. Immunity can also be enhanced by supplementing probiotics to monogastrics diets. Moreover, probiotics can help in improving major meat quality traits and countering a variety of monogastric animals infectious diseases. A proper selection of the probiotic strains is required in order to confer optimal beneficial effects. The present review focuses on the general functional, safety, and technological screening criteria for selection of ideal Bacillus probiotics as feed supplements as well as their mechanism of action and beneficial effects on monogastric animals for improving production performance and health status.
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Palm oil milling has produced tonnes of solid waste including palm decanter cake (PDC). The recalcitrant nature of PDC limits its full inclusion in animal feed. This study aims to investigate the effect of fibre-degrading enzyme such as cellulase and xylanase on the prebiotic activity and solid state fermentability of PDC. We used the following commercial enzyme loading: 5U cellulase (5UC), 5U xylanase (5UX) and combined enzymatic treatment 2.5U:2.5U (C25U) per gram of substrate to hydrolyse the defatted PDC. The sugar profile in the effluent was analysed by using high-performance liquid chromatography, and the degree of hydrolysis (DH) was estimated based on the total carbohydrates amount in the effluent. The DH of enzymatic-hydrolysed PDC followed the order of 5UC< C25U <5UX, which was 7.8 %, 44.2 2% and 46.27 %, respectively. The prebiotic activity score of ethanolic extract obtained from the PDC followed the order of untreated < C25U < 5UX < 5UC, which were -1.04, -0.74, -0.10 and 0.58, respectively. To further investigate the role of lignin (which can be eliminated through alkaline hydrogen peroxide treatment (AHPT) on the extent of hydrolysis and the fermentability of enzymatic-hydrolysed PDC, we tested the invasion capacity of fungus Aspergillus oryzae on untreated, and AHPT followed by enzymatic-treated PDC. Pre-treatment of PDC with AHP improved accessibility for enzymatic hydrolysis in which the highest fungus growth rate was observed on the AHP-C25U PDC. Enzymatic treatment succeeding AHPT is a feasible way to improve the fermentability of palm decanter cake.
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Different permeate-based media were screened for their productivity of Lactobacillus bulgaricus-β-galactosidase. The P4 medium containing yeast extract, NH4CI, K2HPO4, peptone gave the best activity (2-6 u/ml). Supplementation of P4 medium with several test amino acids as well as different test sugars failed to support the enzyme production. On the other hand, (NH4)2HPO4 at a concentration of 0.4-0.5% was the best nitrogen source supporting higher activity reaching 4.28 u/ml. The presence of a buffering system composed of 1g/l K2HPO4, 0.5 g/l KH2PO4 enhanced the enzyme production up to 5.16 u/ml. Pretreatment of milk with β-galactosidase enhanced growth of yoghurt starter bacteria. However, lower quality fermented milk was obtained.
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Colombatto, D. and Beauchemin, K. A. 2003. A proposed methodology to standardize the determination of enzymic activities present in enzyme additives used in ruminant diets. Can. J. Anim. Sci. 83: 559-568. There is increasing interest in using enzymes that degrade plant cell walls in ruminant diets to enhance production efficiency. Despite strong evidence from several studies suggesting a beneficial effect of enzyme supplementation on nutrient utilization and animal performance, overall the results have been somewhat inconsistent. One of the main problems faced by researchers is the lack of adequate biochemical characterization of the products used, which leads to a poor understanding of their mode of action. Of these biochemical characteristics, enzyme activities are the most important, but they are not always evaluated prior to use. Furthermore, as many arbitrary units of expression for these activities coexist, direct comparisons among studies are essentially impossible. In this paper, we propose a methodology that we feel accounts for the requirements of accuracy, simplicity and safety of use. In addition, a rationale for the standardization of the assays as a function of the conditions under which the enzymes are expected to act is presented. The standardization of these assays will benefit researchers, the feed industry, regulatory organizations, and ultimately the consumer, as it will result in the development of better, safer and more consistent enzyme additives for use in ruminant diets.
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The objective of this study was to investigate the effect of method of delivery of a solution contaning cellulase on the in vitro rumen fermentation (24 h) of three tropical forages. Enzyme was applied to forages either at the time of incubation or 24 h before. Both cellulase treatments increased (P<0.05) acetate, propionate and total VFA production, as well as neutral-detergent fibre degradability (NDFD) with all forages. NDFD increased significantly (P<0.05) with all forages when 24 h pre-treatment of forage with the enzyme was allowed, and this pre-treatment also tended to increase (P<0.05) gas production with two of the three forages. The results indicate that 24 h pre-treatment of forages with cellulase could increase the efficacy of this enzyme, but the incubated forage affects the results.