Β-glucanase Activity Research Articles

Comparative metagenomic analysis of microcosm structures and lignocellulolytic enzyme systems of symbiotic biomass-degrading consortia

Decomposition of lignocelluloses by cooperative microbial actions is an essential process of carbon cycling in nature and provides a basis for biomass conversion to fuels and chemicals in biorefineries. In this study, structurally stable symbiotic aero-tolerant lignocellulose-degrading microbial consortia were obtained from biodiversified microflora present in industrial sugarcane bagasse pile (BGC-1), cow rumen fluid (CRC-1), and pulp mill activated sludge (ASC-1) by successive subcultivation on rice straw under facultative anoxic conditions. Tagged 16S rRNA gene pyrosequencing revealed that all isolated consortia originated from highly diverse environmental microflora shared similar composite phylum profiles comprising mainly Firmicutes, reflecting convergent adaptation of microcosm structures, however, with substantial differences at refined genus level. BGC-1 comprising cellulolytic Clostridium and Acetanaerobacterium in stable coexistence with ligninolytic Ureibacillus showed the highest capability on degradation of agricultural residues and industrial pulp waste with CMCase, xylanase, and β-glucanase activities in the supernatant. Shotgun pyrosequencing of the BGC-1 metagenome indicated a markedly high relative abundance of genes encoding for glycosyl hydrolases, particularly for lignocellulytic enzymes in 26 families. The enzyme system comprised a unique composition of main-chain degrading and side-chain processing hydrolases, dominated by GH2, 3, 5, 9, 10, and 43, reflecting adaptation of enzyme profiles to the specific substrate. Gene mapping showed metabolic potential of BGC-1 for conversion of biomass sugars to various fermentation products of industrial importance. The symbiotic consortium is a promising simplified model for study of multispecies mechanisms on consolidated bioprocessing and a platform for discovering efficient synergistic enzyme systems for biotechnological application.

β-Glucanase specific expression in the parotid gland of transgenic mice

The feasibility of using the pig parotid secretory protein promoter to drive the β-glucanase transgene expression in mouse parotid glands was examined in this study. The parotid gland-specific vector expressing β-glucanase gene (GLU, from Paenibacillus polymyxa CP7) was constructed. Transgenic mice were produced by the pronuclear microinjection. Both PCR and Southern blot analysis showed that the mice carried the β-glucanase gene and the β-glucanase gene could be stably inherited. Furthermore, RT-PCR and northern blot analysis indicated that it was specifically expressed in the parotid. The β-glucanase activity in the saliva was found to be 0.18U/mL. After feeding a diet containing 2% β-glucan, the average daily gain of transgenic was significantly higher than non-transgenic mice. The crude protein and crude fat concentration in faeces of transgenic mice were significantly reduced compared with that of the non-transgenic mice. These results suggest that the successful expression of foreign β-glucanase in the animal parotid would offer a promising biological approach to reduce the anti-nutritional effect of β-glucans in feed.

Assessment of potential enhancing effects of a carbohydrase mixture on phytase efficacy in male broiler chicks fed phosphorus-deficient diets from 1 to 18 days of age

A total of 540 one-day-old male broiler chicks were used to evaluate the potential enhancing effects of a mixture of 2 commercial carbohydrase preparations on phytase efficacy. A nonphytate phosphorus (nPP) deficient diet (0.15%) was supplemented with different levels of phytase (0X, 1X, 2X, and 3X of recommended level of 500 phytase units per kg of feed), individually or in combination with different levels of a mixture of 2 commercial carbohydrase enzymes [0X, 1X, and 2X of recommended level (X = 0.2 g per kg of feed; a mix with predominantly xylanase and β-glucanase activity)] to determine their effects on performance and bone development. A standard response curve was developed using graded levels of nPP (0.15 to 0.45%) from dicalcium phosphate in a diet with no phytase supplementation to estimate the phosphorus equivalency of the different enzyme combinations. Each diet was fed to 6 replicate pens of 5 male chicks from 1 to 18 d of age. Phytase at normal usage levels of 500 phytase units per kg of feed was effective (P < 0.05) in improving BW, increasing feed intake, reducing mortality rate, and increasing toe and tibia ash without significantly influencing feed conversion. Addition of extra levels of phytase (2X and 3X) significantly (P < 0.05) improved toe and tibia ash without further affecting performance traits. The carbohydrase mixture failed to improve performance or enhance phytase efficacy. The phosphorus equivalency of phytase was dose dependent, with estimates of 0.100, 0.123, and 0.213% for 1X, 2X, and 3X supplementation levels of phytase, respectively, based on tibia ash. In conclusion, results indicate that increasing the level of phytase resulted in significant improvements in utilization of dietary nPP, whereas the carbohydrase preparation had no significant effects on performance or had limited effects in enhancing phytase ability in young broiler chicks.

Novel sources of β-glucanase for the enzymatic degradation of schizophyllan

Schizophyllan is a homoglucan produced by the fungus Schizophyllum commune, with a β-1,3-linked backbone and β-1,6-linked side chains of single glucose units at every other residue. Schizophyllan is commercially produced for pharmaceutical and cosmetics uses. However, surprisingly little information is available on the biodegradation of schizophyllan. Enzymes that attack schizophyllan could be useful for controlled modifications of the polymer for novel applications. Enrichment cultures were used to isolate 20 novel fungal strains from soil samples, capable of growing on schizophyllan as a sole carbon source. Three additional strains were isolated as contaminants of stored schizophyllan solutions. Strains showing the highest levels of β-glucanase activity were identified as Penicillium simplicissimum, Penicillium crustosum, and Hypocrea nigricans. β-glucanases also showed activity against the similar β-glucans, laminarin and curdlan. By comparison, commercial β-glucanase from Trichoderma longibrachiatum and laminarinase from Trichoderma sp. showed lower specific activities toward schizophyllan than most of the novel isolates. β-glucanases from P. simplicissimum and H. nigricans exhibited temperature optima of 60°C and 50°C against schizophyllan, respectively, with broad pH optima around pH 5.0. Partial purifications of β-glucanase from P. simplicissimum and P. crustosum demonstrated the presence of multiple active endoglucanase species, including a 20–25kD enzyme from P. simplicissimum.

Presence of β-glucanase activity in wheat and dairy ingredients and use of organic salts as potential enzyme inhibitors

Depolymerization of mixed linkage cereal β-glucans has been observed during the mixing and fermentation of multigrain breads. To maintain the bioactivity of β-glucans in bread and other foods, control of β-glucanase activity during production is needed. β-Glucanase activity in wheat and dairy ingredients was determined by adding them to β-glucan solutions and following the change in the molecular weight of β-glucan at 50 °C for 60 min. The activities in six wheat and six dairy ingredients ranged from undetectable to a 98% reduction in molecular weight. Calcium propionate, potassium sorbate, and sodium benzoate are salts of organic acids commonly used as food preservatives. Low concentrations of these additives (0.1–1.0%) were found to have an inhibitory effect on the β-glucanases found in wheat flour and whey protein isolate. The depolymerization rate of β-glucan was reduced by up to 76% by the three additives.

Effect of enzyme addition to forage at ensiling on silage chemical composition and NDF degradation characteristics

The effect of different exogenous fibrolytic enzymes added to forages at ensiling was examined for effect on chemical composition and in vitro NDF degradability characteristics of the resulting silage. Maize stover and lucerne were used to study effect on chemical composition in experiment 1, and two varieties of maize stover, lucerne and grass clover were used to study NDF degradation characteristics in experiment 2. Forages were treated with enzymes (500mg crude protein of the enzyme products/kg DM) and ensiled for 60 days in vacuum-sealed bags. Samples of forage (before ensiling) and silage were analysed for chemical composition and silages were analysed for pH and fermentation products. The in vitro NDF degradation characteristics of four forages treated with selected enzymes were measured by incubation for up to 96h with rumen fluid.Enzymes with glucanase, β-glucanase and pectinase activity increased lactic acid and decreased butyric acid, ammonia and pH compared with control silage, and increased glucose concentration in lucerne silage. NDF concentration generally decreased due to enzyme treatment with glucanase, β-glucanase and xylanase activity and in vitro organic matter digestibility decreased in treated maize stover silage. Potential NDF degradability decreased due to enzyme treatments but not for all maize stover treatments. Treatments with combination of enzymes with glucanase, β-glucanase and pectinase activity mostly resulted in increases in fermentation products compared with treatment with individual enzymes. Enzyme mix with xylanase, glucanase and β-glucanase activities was effective for maize stover, whereas a mix containing pectinase activity was most effective for reducing pH in lucerne.Data from this study suggest that adding fibrolytic enzymes to forages at ensiling can solubilise some of the easily digestible parts of cell wall carbohydrates and thereby supply substrates for lactate fermentation. Therefore enzyme addition could be efficient to improve silage quality in forages which are difficult to ensile due to low sugar concentration and/or high protein concentration and buffer capacity. Further, treatment with the enzymes reduced the NDF concentration and thereby might increase the energy value and enzyme treatment might be beneficial to increase DM digestibility in high producing dairy cattle with short rumen retention time.

Effects of the inclusion of carbohydrases and different soybean meals in the diet on palatability, digestibility and faecal characteristics in dogs

The objectives of this study were to evaluate the palatability, coefficient of total tract apparent digestibility (CTTAD), metabolisable energy (ME) and faecal characteristics of dogs fed diets with increasing levels of soybean meals (SBM) with or without the inclusion of an enzyme complex with α-galactosidase, β-glucanase, and xylanase activity. Iso-proteic diets containing 0, 150 or 300g of regular SBM (RSBM)/kg, corresponding to 460g of crude protein (CP)/kg, or high-protein SBM (HSBM), corresponding to 500gCP/kg, with or without 2.0genzymes/kg, were evaluated in three experiments. In experiment 1, a palatability test was performed to make six pair-wise comparisons: 0 vs. 150g RSBM/kg; 0 vs. 300g RSBM/kg; 300g RSBM vs. 300g HSBM/kg; 0 vs. 0g RSBM/kg+enzymes; 300g RSBM vs. 300g RSBM/kg+enzymes and 300g HSBM vs. 300g HSBM/kg+enzymes. In addition, two digestibility trials were conducted to evaluate three levels of RSBM (experiment 2) or HSBM (experiment 3) diets. The results of experiment 1 indicated that dogs preferred diets containing 300g HSBM/kg to the RSBM diet (P<0.05), whereas they preferred RSBM diets relative to the control diet (P<0.05). Dogs also preferred the control and RSBM diets with enzymes to the diets without enzymes (P<0.05). In experiments 2 and 3, enzyme supplementation increased the CTTAD of dry matter (DM), organic matter and CP of the RSBM and HSBM diets (P<0.05). The RSBM×enzymes interaction supplied 0.53MJ/kg in addition to the ME in the 150g RSBM/kg+enzymes diet, relative to the control diet (P<0.05). The inclusion of RSBM and HSBM in the diet increased the CTTAD of the DM and CP compared to the control diet but reduced that of ether extract and ME (P<0.05). The faecal characteristics of the dogs were not influenced by the enzyme supplementation (P>0.05). An increase in the level of inclusion of RSBM and HSBM reduced the DM content, faecal score (looser stools), and pH, whereas the volume of the faeces produced was increased (P<0.05). Enzyme supplementation increased palatability, the DM and CP digestibility, and the ME content of dog foods containing up to 300g of the two SBM/kg.

Fundamental study on the influence of Fusarium infection on quality and ultrastructure of barley malt

Barley infection with Fusarium species has been a long standing problem for the malting and brewing industries. In this study, we evaluate the impact of Fusarium culmorum infected raw barley on the final malt quality. Barley grains were infected for 5days at optimum fungal growth conditions. Grains were fully characterized and compared to standard barley grains. Due to fungal infection, germinative energy of infected barley grains decreased by 45%; its water sensitivity increased dramatically, and grains accumulated 199μg/kg of deoxynivalenol (DON). Barley grains were subsequently malted for 8days, fully characterized and compared to standard malt grains. Fungal growth behavior was evaluated during malting using a PCR-based assay and mycotoxins were measured using HPLC. Fungal biomass increased in grains, during all stages of malting. Infected malt accumulated 8-times its DON concentration during malting. Kernel ultrastructure was evaluated using scanning electron and confocal laser scanning microscopy. Infected malt grains were characterized by extreme structural proteolytic, (hemi)-cellulolytic and starch deterioration with increased friability and fragmentation. Infected grains had higher protease and β-glucanase activities, lower amylase activity, a greater proportion of free amino and soluble nitrogen, and a lower β-glucan content. Malt loss was over 27% higher in infected malt in comparison to the control. The results of this study revealed that 20% F. culmorum infected barley kernels lead to a significant reduction in malt quality as well as mycotoxin formation.

Isolation and Characterization of Cellulose-decomposing Bacteria Inhabiting Sawdust and Coffee Residue Composts

Clarifying the identity and enzymatic activities of microorganisms associated with the decomposition of organic materials is expected to contribute to the evaluation and improvement of composting processes. In this study, we examined the cellulolytic and hemicellulolytic abilities of bacteria isolated from sawdust compost (SDC) and coffee residue compost (CRC). Cellulolytic bacteria were isolated using Dubos mineral salt agar containing azurine cross-linked (AZCL) HE-cellulose. Bacterial identification was performed based on the sequence analysis of 16S rRNA genes, and cellulase, xylanase, β-glucanase, mannanase, and protease activities were characterized using insoluble AZCL-linked substrates. Eleven isolates were obtained from SDC and 10 isolates from CRC. DNA analysis indicated that the isolates from SDC and CRC belonged to the genera Streptomyces, Microbispora, and Paenibacillus, and the genera Streptomyces, Microbispora, and Cohnella, respectively. Microbispora was the most dominant genus in both compost types. All isolates, with the exception of two isolates lacking mannanase activity, showed cellulase, xylanase, β-glucanase, and mannanase activities. Based on enzyme activities expressed as the ratio of hydrolysis zone diameter to colony diameter, it was suggested that the species of Microbispora (SDCB8, SDCB9) and Paenibacillus (SDCB10, SDCB11) in SDC and Microbispora (CRCB2, CRCB6) and Cohnella (CRCB9, CRCB10) in CRC contribute to efficient cellulolytic and hemicellulolytic processes during composting.

Effect of Exogenous Fibrolytic Enzyme Application on the Microbial Attachment and Digestion of Barley Straw In vitro.

The effects of exogenous fibrolytic enzymes (EFE; a mixture of two preparations from Trichoderma spp., with predominant xylanase and β-glucanase activities, respectively) on colonization and digestion of ground barley straw and alfalfa hay by Fibrobacter succinogenes S85 and Ruminococcus flavefaciens FD1 were studied in vitro. The two levels (28 and 280 μg/ml) of EFE tested and both bacteria were effective at digesting NDF of hay and straw. With both substrates, more NDF hydrolysis (p<0.01) was achieved with EFE alone at 280 than at 28 μg/ml. A synergistic effect (p<0.01) of F. succinogenes S85 and EFE on straw digestion was observed at 28 but not 280 μg/ml of EFE. Strain R. flavefaciens FD1 digested more (p<0.01) hay and straw with higher EFE than with lower or no EFE, but the effect was additive rather than synergistic. Included in the incubation medium, EFE showed potential to improve fibre digestion by cellulolytic ruminal bacteria. In a second batch culture experiment using mixed rumen microbes, DM disappearance (DMD), gas production and incorporation of 15N into particle-associated microbial N (15N-PAMN) were higher (p<0.001) with ammoniated (5% w/w; AS) than with native (S) ground barley straw. Application of EFE to the straws increased (p<0.001) DMD and gas production at 4 and 12 h, but not at 48 h of the incubation. EFE applied onto S increased (p<0.01) 15N-PAMN at 4 h only, but EFE on AS increased (p<0.001) 15N-PAMN at all time points. Prehydrolysis increased (p<0.01) DMD from both S and AS at 4 and 12 h, but reduced (p<0.01) 15N-PAMN in the early stage (4 h) of the incubation, as compared to non-prehydrolyzed samples. Application of EFE to barley straw increased rumen bacterial colonization of the substrate, but excessive hydrolytic action of EFE prior to incubation decreased it.

Malting revisited: Germination of barley (Hordeum vulgare L.) is inhibited by both oxygen deficiency and high carbon dioxide concentrations

During malting, barley (Hordeum vulgare L.) seeds are germinated to promote the mobilisation of storage compounds. Germination is strongly influenced by O2 and CO2; however, any distinction between the particular effects is missing. Since, in this study, the ambient O2 concentration was maintained when high CO2 concentrations were applied, for the first time the impacts of CO2 and of O2 deficiency could be distinguished unambiguously.Germination was inhibited by both O2 deficiency and high CO2 (80%) concentrations, documented by the lack of any growth of coleoptiles and any increase of α-amylase and β-glucanase activity. In contrast, the related impacts of O2 starvation and high CO2 on fermentation differ strongly, demonstrated by quite different patterns of ethanol emission. Additionally, the stress metabolism – monitored by the means of GABA accumulation – was also differently impacted. The elucidation of the underlying, so far unknown, mechanisms will provide novel opportunities to improve malting.

Purification and characterization of cold-active endo-1,4-β-glucanase produced by Pseudoalteromonas sp. AN545 from Antarctica

A bacterium hydrolyzing carboxymethylcellulose, isolated from Antarctic sea ice, was identified as Pseudoalteromonas sp. based on 16S rDNA gene sequences and named as Pseudoalteromonas sp. AN545. The extracellular endo-1,4-β-glucanase AN-1 was purified successively by ammonium sulfate precipitation, DEAE-Sepharose ion exchange chromatography and Sephadex G-75 gel filtration chromatography. The molecular mass of AN-1 was estimated to be 47.5 kDa utilizing SDS-PAGE and gel chromatography analysis. AN-1 could hydrolyze caboxymethylcellulose, avicel and β-glucan, but not cellobiose, xylan and p-Nitrophenyl-β-D-glucopyranoside. The optimal temperature and pH for the β-glucanase activity of AN-1 were determined to be at 30°C and pH 6.0, respectively. AN-1 was stable at acidic solutions of pH 5.0–6.5 and temperatures below 30°C for 1 h. Moreover, the specific activity was enhanced by Ca2+ and Mg2+, and inhibited by Cu2+. The kinetic parameters Michaelis constant (K m) and maximum velocity (V max) of AN-1 were 3.96 mg/mL and 6.06×10−2 mg/(min·mL), respectively.

Levels of endogenous β-glucanase activity in barley affect the efficacy of exogenous enzymes used to supplement barley-based diets for poultry

To improve the nutritive value of barley-based diet for broilers, 2 experiments using 2 different barley lots were performed to evaluate the capacity of a mesophilic cellulase when fused to a β-glucan specific family 11 carbohydrate-binding module. The data revealed that the recombinant β-glucanase derivatives were not appropriate for feed supplementation because of a lack of stability at acidic pH levels. However, under the same experimental conditions, a commercial enzyme mixture improved the nutritive value of 1 of the cereal lots used. Analysis of the nutritive value of the 2 barleys revealed intrinsic differences in the levels of endogenous β-glucanase activity. These differences were extensively evident when the studies were expanded to a range of 64 barley lots. Thus, to clarify the effect of endogenous cellulases on the efficacy of exogenous β-glucanases used to supplement barley-based diets for poultry, 2 barley lots presenting low and high levels of endogenous plant cell wall-degrading enzymes were selected. These lots were used to prepare 2 barley-based diets, which were supplemented with or without a commercial enzyme product and fed to broiler chicks. The data revealed that the exogenous enzymes were effective when the basal diet presented low levels of endogenous β-glucanases but were unable to improve the nutritive value of the barley lot displaying higher β-glucanase activity. Thus, these studies suggest that levels of endogenous β-glucanases may affect the efficacy of exogenous enzymes used to improve the nutritive value of barley-based diets for broilers. The development of a quick β-glucanase assay that could be applied for cereal-based feeds may help identify those barley-based diets that are more responsive to the action of feed enzymes.

Molecular cloning and characterization of a β-glucanase from Piromyces rhizinflatus

Cellulose is the most abundant renewable polysaccharide with a high potential for degradation to useful end products. In nature, most cellulose is produced as crystalline cellulose. Therefore, cellulases with high hydrolytic activity against crystalline cellulose are of great interest. In this study, a crystalline cellulose degradation enzyme was investigated. The cDNA encoding a β-glucanase, CbhYW23-2, was cloned from the ruminal fungus Piromyces rhizinflatus. To examine the enzyme activities, CbhYW23-2 was expressed in Escherichia coli as a recombinant His(6) fusion protein and purified by immobilized metal ion-affinity chromatography. Response surface modeling (RSM) combined with central composite design (CCD) and regression analysis was then employed for the planned statistical optimization of the β-glucanase activities of CbhYW23-2. The optimal conditions for the highest β-glucanase activity of CbhYW23-2 were observed at 46.4°C and pH 6.0. The results suggested that RSM combined with CCD and regression analysis were effective in determining optimized temperature and pH conditions for the enzyme activity of CbhYW23-2. CbhYW23-2 also showed hydrolytic activities toward Avicel, carboxymethyl cellulose (CMC), lichenan, and pachyman. The results also proved that the high activity of CbhYW23-2 on crystalline cellulose makes it a promising candidate enzyme for biotechnological and industrial applications.

Evaluation of Cellulolytic and Hemicellulolytic Abilities of Fungi Isolated from Coffee Residue and Sawdust Composts

This study focused on the evaluation of cellulolytic and hemicellulolytic fungi isolated from sawdust compost (SDC) and coffee residue compost (CRC). To identify fungal isolates, the ITS region of fungal rRNA was amplified and sequenced. To evaluate enzyme production, isolates were inoculated onto wheat bran agar plates, and enzymes were extracted and tested for cellulase, xylanase, β-glucanase, mannanase, and protease activities using different azurine cross-linked (AZCL) substrates. In total, 18 isolates from SDC and 29 isolates from CRC were identified and evaluated. Four genera (Aspergillus, Galactomyces, Mucor, and Penicillium) and five genera (Aspergillus, Coniochaeta, Fusarium, Penicillium, and Trichoderma/Hypocrea) were dominant in SDC and CRC, respectively. Penicillium sp., Trichoderma sp., and Aspergillus sp. displayed high cellulolytic and hemicellulolytic activities, while Mucor isolates exhibited the highest β-glucanase and mannanase activities. The enzyme analyses revealed that Penicillium, Aspergillus, and Mucor isolates significantly contributed to the degradation of SDC, whereas Penicillium, Aspergillus, and Trichoderma isolates had a dominant role in the degradation of CRC. Notably, isolates SDCF5 (P. crustosum), CRCF6 (P. verruculosum), and CRCF2 and CRCF16 (T. harzianum/H. lixii) displayed high activity regarding cellulose and hemicellulose degradation, which indicates that these species could be beneficial for the improvement of biodegradation processes involving lignocellulosic materials.

Genetic variants of HvGlb1 in Tibetan annual wild barley and cultivated barley and their correlation with malt quality

Improvement of malt quality is the most important objective in malt barley breeding. The current experiments investigated the variation of malt quality characters among barley genotypes and the difference in genetic variants of HvGlb1, encoding β-glucanase isoenzyme I, between Tibetan annual wild barley and cultivated barley. The correlation between the gene variants and malt quality showed that there was a large difference in the four malt quality parameters, i.e. Kolbach index, diastatic power (DP), viscosity and malt extract, among the analyzed barley cultivars. Kolbach index was negatively and positively correlated with viscosity and malt extract, respectively, while malt extract was negatively correlated with viscosity. Malt β-glucan content was a major determinant of malt quality, and was significantly correlated with Kolbach index (−0.633), malt extract (−0.333) and viscosity (0.672). On the other hand, malt β-glucan content was mainly controlled by malt β-glucanase activity. The correlation analysis showed that the HvGlb1 gene was correlated with malt β-glucan content and three of four main malt quality parameters, except DP. In addition, we also found that the HvGlb1 of Tibetan barley had wider diversity in haplotype than that of the cultivated barley, supporting the hypothesis that Tibet is one of the original centers of cultivated barley.

Characterization of Glycoside Hydrolase Family 5 Proteins in Schizosaccharomyces pombe

In yeast, enzymes with β-glucanase activity are thought to be necessary in morphogenetic events that require controlled hydrolysis of the cell wall. Comparison of the sequence of the Saccharomyces cerevisiae exo-β(1,3)-glucanase Exg1 with the Schizosaccharomyces pombe genome allowed the identification of three genes that were named exg1(+) (locus SPBC1105.05), exg2(+) (SPAC12B10.11), and exg3(+) (SPBC2D10.05). The three proteins have different localizations: Exg1 is secreted to the periplasmic space, Exg2 is a membrane protein, and Exg3 is a cytoplasmic protein. Characterization of the biochemical activity of the proteins indicated that Exg1 and Exg3 are active only against β(1,6)-glucans while no activity was detected for Exg2. Interestingly, Exg1 cleaves the glucans with an endohydrolytic mode of action. exg1(+) showed periodic expression during the cell cycle, with a maximum coinciding with the septation process, and its expression was dependent on the transcription factor Sep1. The Exg1 protein localizes to the septum region in a pattern that was different from that of the endo-β(1,3)-glucanase Eng1. Overexpression of Exg2 resulted in an increase in cell wall material at the poles and in the septum, but the putative catalytic activity of the protein was not required for this effect.

Effects of Supplemental Enzymes in Diets Containing Distillers Dried Grains with Solubles on Finishing Pig Growth Performance

ABSTRACT Four experiments involving 4,506 pigs were conducted to determine the effects of different commercial enzymes on the growth performance of growing-finishing pigs fed distillers dried grains with solubles (DDGS). All experiments were conducted in the same commercial swine research facility. Experiments 1 and 2 used corn- and soybean meal-based diets with 15% DDGS. A β-mannanase enzyme was used in Exp. 1, and a blend of enzymes that contained β-glucanase, cellulase, and protease activities was used in Exp. 2. There were no differences (P > 0.10) in ADG, ADFI, and G:F between pigs fed diets with added enzyme and pigs fed diets without enzyme in either experiment (ADG 1.00 vs. 1.01 kg/d, and G:F 0.408 vs. 0.408; ADG 0.94 vs. 0.94 kg/d, and G:F 0.424 vs. 0.421, in Exp. 1 and 2, respectively) In Exp. 3, diets containing 45 and 60% DDGS were fed with or without 2 commercial proprietary enzyme products designed for use in diets containing DDGS. Average daily gain (0.85 vs. 0.86 kg/d) and G:F (0.370 vs. 0.373) were not different (P > 0.10) among treatments. In Exp. 4, an enzyme product with bacterial endo-1,4-β-xylanase was evaluated in diets containing 30% DDGS. Average daily gain (0.82 vs. 0.82 kg/d) and G:F (0.391 vs. 0.387) were not different (P > 0.10) among treatments. Based on the results, the different enzymes evaluated in these experiments did not enhance finishing pig growth performance when diets contained varying levels of DDGS.

Analysis of grain yield and cytolytic degradation of winter and spring barley cultivars.

Six barley cultivars were grown in locations of Osijek and Nova Gradiska (Croatia) for three successive years. Our study included cultivars of different seasonal type. ‘Zlatko’, ‘Barun’ and ‘Vanessa’ are two-rowed winter type barley cultivars and ‘Fran’, ‘Matej’ and ‘Scarlett’ are spring barley cultivars. The results of grain yield and cytolytic degradation showed significant differences among winter and spring cultivars under different growing conditions. Winter type cultivars ‘Zlatko’ and ‘Barun’ showed significantly higher grain yield in comparison with other cultivars. Generally, spring barley cultivars when compared with winter cultivars had significantly higher malt extract content, lower malt extract fine-coarse difference, lower malt viscosity, higher β-glucanase activity and lower malt β-glucan content, higher friability.

Comparative assessment of amylolytic and cellulolytic enzyme activity of malts prepared from tropical cereals

The activity levels of amylolytic enzymes and β-glucanase in malts prepared from four tropical cereal grains were assessed to establish the relative usefulness of these malts for production of glucose syrups. Rice malt showed the highest activity for the amylolytic enzymes, whilst millet and sorghum malts were richest in β-glucanase activity. Optimum amylolytic enzyme development in rice malts occurred between 9–13 days; and 11 days for optimum β-glucanase development in millet malt. β-Amylase was the predominant enzyme in all the cereal malts except maize, for which the predominant enzyme was α-amylase. Options for optimising the production of enzymes from the three cereals, and their potential for use in the production of glucose syrups are discussed.