Endogenous Xylanase Research Articles

Using in feed xylanase or stimbiotic to reduce the variability in corn nutritive value for broiler chickens

This study investigated the effects of xylanase and stimbiotic (fiber fermentation enhancer) on the response of broiler chickens fed different corn varieties and determine correlations between variables of interest. Four corn genetic varieties were selected based on their range in nutrient composition. Diets containing 600 g/kg of each corn were supplemented with 0 or 100 g/ton of xylanase or stimbiotic. A total of 1,152 one-day-old male broiler chicks (Ross 308) were divided into 12 treatments, each with 8 pens and 12 birds per pen, for a 21-day study. On d 21, performance parameters were measured, and the ileal energy and organic matter (OM) digestibility and cecal xylanase activity determined. Stimbiotic supplementation improved mFCR compared with all other treatments. There was a treatment by corn variety interaction for body weight (BW), BW gain and coefficient of variation (CV) of BW (P ≤ 0.05). Birds fed corn Variety 1 (highest neutral dietary fiber, protein and soluble arabinoxylan content) supplemented with stimbiotic had the highest BW, while Variety 2 control diet had the lowest. The BW CV in corn Variety 2 was the highest, which improved with stimbiotic supplementation. The BW CV in corn Variety 1 responded better to stimbiotic than xylanase. There were no treatment differences on BW CV in corn Varieties 3 and 4. The lowest OM digestibility was observed in birds fed corn Variety 1 with xylanase, and the highest value was associated with corn Variety 3 with xylanase (highest total arabinoxylan). Xylanase and stimbiotic supplementation increased the endogenous xylanase activity regardless of the corn variety (P ≤ 0.05). Positive correlations between corn fiber contents and phytic acid and the arabinose:xylose ratio were seen, while nonstarch polysaccharide content was negatively correlated with apparent metabolizable energy. In conclusion, corn variety influenced nutrient digestibility and broiler chicken growth. The response to supplementation with xylanase or stimbiotic varied based on the nutritional profile of corn with regards to improvements in digestibility and performance in broiler chickens.

Enhanced expression of xylanase in Aspergillus niger enabling a two-step enzymatic pathway for extracting β-glucan from oat bran

The high cost and process complexity limit the enzymatic extraction of β-glucan. In this study, β-glucan was extracted from oat bran in a two-step enzymatic pathway using a recombinant strain of Aspergillus niger AG11 overexpressing the endogenous xylanase (xynA) and amylolytic enzyme. First, co-optimization of promoter and signal peptide and a fusion of glucoamylase (glaA) fragment were integrated into the β-glucosidase (bgl) locus to improve xynA expression. Then, the optimized expression cassette was simultaneously integrated into bgl, α-amylase amyA, and acid α-amylase ammA loci, yielding the Rbya with 3,650-fold and 31.2% increase in xynA and amylolytic enzyme activity than the wild-type strain, respectively. Finally, Rbya’s supernatants at 72 h (rich in xynA and amylolytic enzyme) and 10 d (rich in proteases) were used to decompose xylan/starch and proteins in oat bran, respectively, to obtain 85.1% pure β-glucan. Rbya could be a robust candidate for the cost-effective extraction of β-glucan.

Characterization of Two Wheat-Derived Glycoside Hydrolase Family-10 Xylanases Resistant to Xylanase Inhibitors

Xylanase inhibitors inhibit the activities of microbial xylanases and seriously compromise the efficacy of microbial xylanases added to modify cereals. Cereal endogenous xylanases are unaffected by these xylanase inhibitors, but little information is available regarding their effects in improving cereal quality, a neglected potential application. As a strategy for circumventing the negative effects of xylanase inhibitors, the objective of this study was to use genetic engineering to obtain sufficient amounts of active endo-1,4-β-D-xylanase from wheat to analyze the characteristics of its structure. The endo-1,4-β-D-xylanase from wheat was heterologously expressed. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), western blotting, MALDI-TOF/TOF (MS) analyses, and enzyme activity determination confirmed 2 active endo-1,4-β-D-xylanases (EXY3 and EXY4) were successfully obtained. The molecular weights (MW) and isoelectric point (pI) of EXY3 were 36.108 kDa and 5.491, while those of the EXY4 protein were 41.933 kDa and 5.726. They both contained the same catalytic domain of GH10 xylanases from G266 to V276 and have the same catalytic site, Glu273. They shared the same putative N-glycosylation sites (N62-T63-S64 and N280–V281–S282) and 3 putative O-glycosylation sites (Ser8, Ser9, and Thr21), but EXY4 had an additional O-glycosylation site (Thr358). EXY3 was smaller than EXY4 by 51 amino acids because of a nonsense mutation and premature termination. They both had the 8-fold beta/alpha-barrel (TIM-barrel) fold. The specific activities of EXY3 and EXY4 were 152.0891 and 67.2928 U/mg, respectively. This work demonstrates a promising way to obtain wheat xylanases by genetic engineering; the properties of the enzymes indicate their potential application in cereal-based industries.

Effect of Bran Pre-Treatment with Endoxylanase on the Characteristics of Intermediate Wheatgrass (Thinopyrum intermedium) Bread.

Previous work indicated that bran removal promotes network formation in breads prepared from intermediate wheatgrass (IWG) flour. However, refinement reduces yields as well as contents of nutritionally beneficial compounds such as fiber. This study evaluated xylanase pretreatment of IWG bran as a processing option to enhance the properties of bread made with half of the original bran content. Xylanase pretreatment did not affect stickiness but significantly reduced hardness and increased specific loaf volumes compared to negative (without xylanase) and positive controls (with xylanase but without pretreatment). However, the surface of breads with pretreated bran was uneven due to structural collapse during baking. Fewer but larger gas cells were present due to pretreatment. Addition of ascorbic acid modulated these effects, but did not prevent uneven surfaces. Accessible thiol concentrations were slightly but significantly increased by xylanase pretreatment, possibly due to a less compact crumb structure. Endogenous xylanases (apparent activity 0.46 and 5.81 XU/g in flour and bran, respectively) may have been activated during the pretreatment. Moreover, Triticum aestivum xylanase inhibitor activity was also detected (193 and 410 InU/g in flour and bran). Overall, xylanase pretreatment facilitates incorporation of IWG bran into breads, but more research is needed to improve bread appearance.

Barley α-amylase/subtilisin inhibitor shows inhibitory activity against endogenous xylanase isozyme I of malted barley: A novel protein function.

Mashing process had little influence on the arabinoxylan content in the finished wort. In this paper, a protein with inhibitory activity against the endogenous xylanase isozyme I (X-I) of malted barley was extracted and purified using a combination of ion-exchange and size-exclusion chromatography. The protein was identified as barley α-amylase/subtilisin inhibitor (BASI). According to the amino acid sequence analysis, BASI was completely different from the previous reported xylanase inhibitors. BASI showed dosage-dependent inhibitory activity. BASI exhibited a maximum inhibitory activity at 50°C and pH 6.0. BASI inhibited X-I as a competitive manner. PRACTICAL APPLICATIONS: A protein with inhibitory activity against the major endogenous xylanase isozyme I (X-I) of malted barley was extracted, purified, and characterized, which was identified as barley α-amylase/subtilisin inhibitor (BASI). The results help brewers to achieve a better understanding of the mechanism of arabinoxylan degradation during mashing. BASI can be used as an indicator to screen microbial xylanases. The microbial xylanases insensitive to BASI would have obvious advantages in the degradation of arabinoxylan polymers and filterability improvement during mashing.

Functional characterization of a new pathogen induced xylanase inhibitor (RIXI) from rice

Rice xylanase inhibitor RIXI is a XIP-type inhibitor that belongs to the glycoside hydrolase family 18 (GH18), which includes plant class III chitinases (EC 3.2.1.14) known as PR-8 proteins. The aim of this study was to elucidate whether RIXI had any effect on rice endoxylanase and its role(s) in plant defence. RIXI encoding sequence was cloned from rice (Oryza sativa cv. Nipponbare), and was expressed in Escherichia coli. The activity of recombinant RIXI was investigated – among the tested xylanases, the GH11 xylanase from Aspergillus niger was the most inhibited, while rice endogenous xylanase OSX was least inhibited. Semi-quantitative real-time polymerase chain reaction (PCR) and quantitative real-time PCR showed that the xylanase inhibitor gene RIXI was up-regulated in rice seedlings infected by Magnaporthe grisea. The increased RIXI expression was also accompanied by significantly elevated expression of pathogenesis-related protein gene (PR-1) and class III chitinase gene (Cht-3). This suggested that RIXI might be involved in environmental responses such as defense against phytopathogens.

Search for Cell-Wall-Degrading Enzymes of World-Wide Rice Grains by PCR and Their Effects on the Palatability of Rice

Such rice cultivars as Japonica, Japonica-Indica hybrid, Javanica and Indica, were evaluated for their main chemical components (amylose content and protein content), pasting property of rice flour (consistency), physical property of the cooked rice grains (adhesion, L3), and enzyme activities (cellulase and xylanase). The amylose content, cellulase activity and xylanase activity showed significant positive or negative correlation with the pasting property (consistency) of rice flour (r = 0.89, r = 0.58, r = 0.70, respectively) and with the physical property of the cooked rice grains (adhesion, L3: r = -0.51, r = -0.61, r = -0.71, respectively) at the level of 1%. Endogenous xylanase and cellulase played important roles to determine the texture of the cooked rice grains similarly to the amylose content. Part of the DNA sequences of the α-glucosidase gene differed among the Japonica, Japonica-Indica hybrid and Indica subspecies. We found discriminative DNA bands appearing by PCR, corresponding to 1,4-β-xylanase and endo-1,4-β-glucanase 13 in the case of Indica rice, Indica-Japonica hybrid rice, and Javanica rice (non-Japonica subspecies). The equation for estimating the physical property (adhesion) of cooked rice grains by PCR was improved by adding novel primers related to the cell-wall-degrading enzymes.

Effects of an exogenous xylanase gene expression on the growth of transgenic rice and the expression level of endogenous xylanase inhibitor gene RIXI

Xylanases have attracted considerable interest in recent years owing to their various applications in industry and agriculture. The use of transgenic plants to produce xylanases is a less expensive alternative to biotechnological programmes. The aim of this study was to elucidate whether introducing a foreign xylanase gene ATX into rice had any adverse effect on plant growth and development. A recombinant xylanase gene ATX was introduced into rice variety Zhonghua 11 through Agrobacterium-mediated transformation. The T₂ generation of transgenic rice was compared with the control (non-transgenic plants). Exogenous xylanase gene ATX was expressed in rice, and all examined transgenic lines exhibited xylanase activity. The transgenic lines (T₂, 'X1-3' and 'X2-5') appeared to grow and develop normally. There were no differences in net photosynthetic rate between transgenic rice lines ('X1-3' and 'X2-5') and wild type (WT) rice plants at the heading/flowering stage. Xylanases are key enzymes in the degradation of plant cell walls. Cell wall composition analysis showed that that there were no changes in cell wall polysaccharides in the root apex but some alterations in leaves in transgenic rice plants. The results also showed that the expression of exogenous xylanase gene ATX in rice would increase the expression of endogenous xylanase inhibitor gene RIXI, which could play a role in plant defence. Thus the stress resistance of transgenic rice plants might be improved. Exogenous xylanase gene ATX could be successfully expressed in rice, and the exogenous protein had no apparent harmful effects on growth and development in transgenic rice plants.

Fermented Wheat Bran as a Functional Ingredient in Baking

ABSTRACTThe aim of the current study was to identify factors influencing the technological functionality of fermented bran. The influences of fermentation type and type of wheat bran on the microbial community, bioactivity, arabinoxylans (AX), and activity of xylanases were studied in the bran ferments. Furthermore, technological quality of ferments was established by using them to replace wheat in baking with a 20% substitution level. Solubilization of AX and endogenous xylanase activity of bran were influenced by the type of bran, fermentation type, and conditions. Peeled bran had a clearly reduced microbial load and different microbial community in comparison to native bran. Bran from peeled kernels contained 10‐fold lower activities of endogenous xylanases in comparison to native bran. Yeast fermentation of bran from peeled kernels increased the level of folates (+40%), free phenolic acids (+500%), and soluble AX (+60%). Bread containing yeast‐fermented peeled bran had improved volume (+10–15%) and crumb softness (25–35% softer) in comparison to unfermented counterparts. Solubilization of AX during the 20 hr fermentation and decreased endogenous xylanase activity are proposed as the main reasons for the improved technological functionality of fermented bran.

Physicochemical properties of arabinoxlans in refrigerated dough

Abstract Dough syruping is considered a problem associated to the quality of refrigerated dough products. Degradation of arabinoxylans (AX) by endogenous xylanase is one of the causes for the dough syruping development. The objective of this study was to determine physicochemical properties of AXs during storage in relation to the endoxylanase activity. Arabinoxylans were extracted from the refrigerated dough samples that were prepared using two different hard red spring varieties, Glenn and Parshall. The changes in weight-averaged molecular weight, the monosaccharide composition and the viscosity of AXs were investigated during refrigeration of dough. Structural features of AX changed dramatically during storage, resulting in changes in the rheological properties. The high molecular weight fraction of AXs was decreased up to 17% after 34 days of storage. Major structural change occurred in the unsubstituted xylose residues, which was detected by Proton Nuclear Magnetic Resonance ( 1 H NMR). The increase in the peak of unsubstituted xylose backbone indicated that mono- and di-substituted AXs have been degraded during the storage. The viscosity of water extractable AX declined by as much as 43%. Linear regression coefficient, R 2 , was ~−0.9 between A/X and dough syruping for both Glenn and Parshall.

Accumulated Evidence Substantiates a Role for Three Classes of Wheat Xylanase Inhibitors in Plant Defense

Since their discovery in 1997, work on proteinaceous wheat xylanase inhibitors (XIs) led to the identification and thorough biochemical and structural characterization of three classes of XIs, namely TAXI-type (Triticum aestivum xylanase inhibitor), XIP-type (xylanase inhibiting protein), and TLXI-type (thaumatin-like xylanase inhibitor) XIs. Already early on, a role in plant defense has been put forward for these proteins, mainly based on the observation that these XIs are only active against xylanases of microbial origin and can not inhibit the plant endogenous xylanases known so far. Considerable effort has been devoted to substantiate this plant defense hypothesis. Data at the genomic, transcriptomic and proteomic levels increasingly provide evidence that XIs, occurring as large polymorphic families, do indeed participate in plant defense. This review summarizes the current knowledge on XIs and the accumulated evidence on their role in plant defense. TLXI and XIP can be classified as pathogenesis-related (PR) proteins of classes PR-5 and PR-8, respectively, based on their homology with thaumatin-like proteins and chitinases. For TAXI, more evidence on the induction of TAXI proteins by a pathogen in tissues that normally do not express TAXI is required before it can be categorized as a new class of PR proteins. At the moment, TAXI proteins should be referred to as inducible defense-related proteins.

Putative endogenous xylanase from brackish-water clam Corbicula japonica

Xylan digestion by Corbicula japonica was investigated according to the hypothesis that C. japonica can breakdown xylan as well as cellulose. C. japonica showed relatively high xylanase activity compared to other bivalve species. Molecular cloning of a xylanase gene was performed in order to determine whether C. japonica possesses an endogenous xylanase, and resulted in the isolation of cDNA with an ORF of 2523 bp corresponding to 840 amino acids (CjXyn10A). CjXyn10A has a Glycoside Hydrolase Family 10 (GHF10) catalytic domain, N-terminal family 4 carbohydrate binding domain and novel C-terminal cysteine-rich domain. Phylogenetic analysis implies that this gene has common evolutional origin with other GHF10 genes of animal origin. Reverse transcription (RT)-PCR analysis and in situ hybridization revealed that CjXyn10A is likely to be expressed in the secretory cells in the digestive gland, suggesting that this enzyme is produced in the same site as previously reported endogenous cellulases of C. japonica. These findings suggest that CjXyn10A is an endogenous xylanase gene of this species. The occurrence of an endogenous xylanase gene in addition to cellulase genes in C. japonica strongly supports our hypothesis that this species can decompose plant-derived structural polysaccharides.

RNAi-Mediated Knockdown of the XIP-Type Endoxylanase Inhibitor Gene, OsXIP, Has No Effect on Grain Development and Germination in Rice

OsXIP (Oryza sativa xylanase inhibitor protein) is a XIP-type xylanase inhibitor which was identified as a protein encoded by a wound stress-responsive gene in rice. Although the OsXIP gene was specifically expressed in mature grains under basal conditions, recombinant OsXIP had no effect on rice endogenous xylanases, and OsXIP-suppressed transgenic rice plants did not exhibit any change in grain development and germination, suggesting that rice development may be independent of OsXIP. Analysis using an OsXIP-specific antibody revealed that OsXIP is markedly accumulated in apoplast in rice root cells by wounding. These results reinforced the possibility that OsXIP is involved in plant defense mechanisms against phytopathogens.

Variability of polymorphic families of three types of xylanase inhibitors in the wheat grain proteome

Cereals contain proteinaceous inhibitors of endo-beta-1,4-xylanases (E.C.3.2.1.8, xylanases). Since these xylanase inhibitors (XIs) are only active against xylanases of microbial origin and do not interact with plant endogenous xylanases, they are believed to act as a defensive barrier against phytopathogenic attack. So far, three types of XIs have been identified, i.e. Triticum aestivum XI (TAXI), xylanase inhibiting protein (XIP), and thaumatin-like XI (TLXI) proteins. In this study the variation in XI forms present in wheat grain was elucidated using high-resolution 2-DE in combination with LC-ESI-MS/MS and biochemical techniques. Reproducible 2-DE fingerprints of TAXI-, XIP-, and TLXI-type XIs, selectively purified from whole meal of three European wheat cultivars using cation exchange chromatography followed by affinity chromatography, were obtained using a pH-gradient of 6 to 11 and a molecular mass range of 10 to 60 kDa. Large polymorphic XI families, not known to date, which exhibit different pI- and/or molecular mass values, were visualised by colloidal CBB staining. Identification of distinct genetic variants by MS/MS-analysis provides a partial explanation for the observed XI heterogeneity. Besides genetic diversity, PTMs, such as glycosylation, account for the additional complexity of the 2-DE patterns.

Effect of wheat type grinding, heat treatment and phytase supplementation on growth efficiency and nutrient utilization of wheat-based diets for broilers

This experiment studied the effect of phytase and processing (heat treatment, grind size) on performance and nutrient utilization of broilers fed diets from 1 to 21 d that included hardred spring (HRS) and durum wheat with three grind sizes (470, 560, 630 µm) with or without heat treatment (oven dry at 80°C for 15 h) before supplementation with phytase (Natuphos® 0 or 600 FTU kg-1). All 24 diets contained 0.5 g Avizyme 1302 kg-1 and were formulated (80% wheat, 20% basal diet) to supply nutrients necessary to meet the requirements of broiler chicks, except for a low level of nonphytate phosphorus (P, 2.7 g kg-1). The medium ground HRS-based diet produced significantly better feed conversion ratio (FCR) and body weights compared with the other two grind sizes. Heat treatment significantly improved feed conversion ratio and apparent metabolizable energy (AME), protein, and P digestibility of diets. It also significantly decreased feed intake, but produced no differences in 21 d body weight. Supplementation with phytase significantly improved AME, protein, and P digestibility of test diets. The AME of diets was influenced by wheat type, processing and grind size. Phytase increased N and P digestibility. Improvements in N and P digestibility by phytase and heat treatment were dependent on wheat type and grind size.The present results suggest an advantage of both phytase and xylanase in durum- and HRS-based diets. Heat processing destroyed endogenous phytase and xylanase in wheat-based diets, and increased the levels of soluble non-starch polysaccharides. Combined phytase, heat treatment, and grind size may synergistically improve N and P digestibility. Key words: Wheat source, grind size, heat treatment, phytase, digestibility, broiler

A xylose-tolerant β-xylosidase from Paecilomyces thermophila: Characterization and its co-action with the endogenous xylanase

An extracellular β-xylosidase from the thermophilic fungus Paecilomyces thermophila J18 was purified 31.9-fold to homogeneity with a recovery yield of 2.27% from the cell-free culture supernatant. It appeared as a single protein band on SDS–PAGE with a molecular mass of approx 53.5 kDa. The molecular mass of β-xylosidase was 51.8 kDa determined by Superdex 75 gel filtration. The enzyme was a glycoprotein with a carbohydrate content of 61.5%. It exhibited an optimal activity at 55 °C and pH 6.5, respectively. The enzyme was stable in the range of pH 6.0–9.0 and at 55 °C. The purified enzyme hydrolyzed xylobiose and higher xylooligosaccharides but was inactive against xylan substrates. It released xylose from xylooligosaccharides with a degree of polymerization ranging between 2 and 5. The rate of xylose released from xylooligosaccharides by the purified enzyme increased with increasing chain length. It had a K m of 4.3 mM for p-nitrophenol-β- d-xylopyranoside and was competitively inhibited by xylose with a K i value of 139 mM. Release of reducing sugars from xylans by a purified xylanase produced by the same organism increased markedly in the presence of β-xylosidase. During 24-hour hydrolysis, the amounts of reducing sugar released in the presence of added β-xylosidase were about 1.5–1.73 times that of the reaction employing the xylanase alone. This is the first report on the purification and characterization of a β-xylosidase from Paecilomyces thermophila.

Wheat value: improvements by feed technology, plant breeding and animal genetics

Wheat is a common ingredient in poultry diets, especially in Europe. Due to its low protein and high starch levels, wheat is essentially used as an energy supplier. Nutritional values of wheat samples vary according to their nutrient content and digestibilities. Variations of nutrient contents may be associated with cultivars and environmental conditions. Digestibility variations in growing chicks may be associated with these factors, and also, with technological treatments of diets and genetic origin of birds.Total nutrient content (starch, protein, lipids and sugars) is strongly related to fibre content through a negative relationship. A large part of fibre content variations of wheat samples depends on their cultivar origin. Wheat protein content depends both on cultivar and environmental conditions. High yield cultivars are often associated with low protein content.Lipid and starch digestibilities of wheat diets given to growing birds may sometimes be rather low.The main reason of the low lipid digestibility values observed with some wheat samples is the high viscosity of water-extract induced by their water-soluble arabinoxylans. This viscosity results from a combination of several variables including the potential applied viscosity value (PAV as mL/g) of the wheat cultivar, the endogenous and exogenous xylanases, and the endogenous anti-xylanase contents. Environmental conditions and technological treatments are major factors acting on xylanase and anti-xylanase contents.A part of wheat starch digestibility variations may be related to accessibility problems in coarse particles due to hard wheat cultivars and coarse grinding. However, coarse particles may be positive for stimulating feed intake of mash diets, and protecting against intestinal transit disorders.In growing chicks, genetic origin of birds may result in very large variations in the digestibility of wheat diets, as observed with the divergent D+ and D- “Digestion” lines selected on the digestion ability of a wheat diet. In 3 w old chickens selected over 4 generations, the AMEn value of a Rialto wheat diet was 13% lower in D- than in D+ birds (P=0.0001). D- birds showed 10% variations in AMEn values between four cereal diets (P=0.0003), while D+ birds showed only 4% variations (P=0.0006). In D+ birds, AMEn values of wheat and maize diets were the same (14,488 vs. 14,538 J/g DM), while, in D- birds, AMEn values of wheat diets were 5% lower (P=0.015) than the maize diet value (13,106 vs. 13,809 J/g DM).

Evidence for the Involvement of Arabinoxylan and Xylanases in Refrigerated Dough Syruping

The relationship between syruping in refrigerated doughs upon prolonged storage and different aspects of arabinoxylan (AX) hydrolysis was investigated using Triticum aestivum xylanase inhibitor (TAXI) and different xylanases in the dough formula. Dough characteristics were evaluated with strong emphasis on the AX population and its fate as a function of storage time. Selective reduction of part of the flour endogenous xylanase activity in dough by added TAXI reduced dough syruping after 12 and 20 days of storage by 50%, providing straightforward evidence for the involvement of xylanases and, thus, AX in the syruping phenomenon. Addition of xylanases with different inhibitor sensitivities [an inhibition-sensitive Bacillus subtilis xylanase (XBS(i)) as well as a noninhibited mutant (XBS(ni)) thereof] to dough confirmed the importance of xylanases in dough syruping, on one hand, and the power of wheat flour TAXI to constitute a significant barrier against xylanase-mediated dough syruping, on the other hand. Use of xylanases with different substrate selectivities [an Aspergillus aculeatusxylanase (XAA) versus XBS(ni)] showed degradation of water-extractable AX (WE-AX) and solubilized AX to low molecular weight molecules rather than the conversion of water-unextractable AX (WU-AX) to high molecular weight water extractable components to be the main factor influencing dough syruping.

Evaluation of novel fungal cellulase preparations for ability to hydrolyze softwood substrates – evidence for the role of accessory enzymes

Seven fungal cellulase preparations from Trichoderma and Penicillium spp. were evaluated for their ability to hydrolyze the cellulose component of lignocellulose samples prepared from softwoods by steam explosion or organosolv pretreatment. Hydrolysis was quantified using two indices: mean specific rate and specific conversion. The activities of the cellulase preparations on model cellulosic substrates (filter paper, carboxymethylcellulose and Avicel) were also determined, together with their β-glucosidase, xylanase, pectinase, galactomannanase and β-glucanase activities. The ability of a cellulase preparation to hydrolyze pretreated softwood showed little correlation with its activity on filter paper, carboxymethylcellulose and Avicel; however, there was a significant correlation with its level of endogenous β-glucosidase and xylanase activity. Differences in the performance of the various cellulase preparations were substantially reduced following supplementation with a commercial β-glucosidase preparation from Aspergillus niger. In addition to β-glucosidase, this preparation was shown to contain significant levels of endogenous xylanase activity. It appears that the levels of endogenous β-glucosidase and xylanase are important factors in determining the ability of a cellulase preparation to hydrolyze pretreated softwood and that deficiencies in the levels of both enzymes can be compensated by supplementation with corresponding activities present in the β-glucosidase preparation. Presumably, β-glucosidase improves cellulose hydrolysis by reducing end-product inhibition by cellobiose, while xylanase activity increases the accessibility of cellulose to cellulases.

Refrigerated dough syruping in relation to the arabinoxylan population.

Refrigerated doughs develop syruping upon prolonged storage. To assess the role of arabinoxylans (AX), in this phenomenon, the evolution of the AX population and syruping in refrigerated doughs during storage were studied. When doughs were kept at 6 degrees C for up to 34 days of storage, dough syruping increased from 0% (fresh dough) to 22% of dough weight, reaching a plateau after 16 days of storage. High-performance size exclusion chromatography and gas-liquid chromatography showed hydrolysis of water-unextractable AX in the refrigerated dough, resulting in increased levels of solubilized AX in the first 2 days of storage. Longer storage resulted in further degradation of solubilized and water-extractable AX. Increased syruping was accompanied by a decrease in farinograph dough consistency. The results support the hypothesis that loss of water-holding capacity due to degradation of AX by endogenous xylanases is responsible for dough syruping.