Cellulolytic Enzyme Preparations Research Articles

Protein preparations from rapse processing waste: A review of the current status and development prospects of existing technologies

The demand for protein products is increasing due to the demographic growth of the world’s population. As an alternative to traditional sources of protein, waste from plant raw material processing is becoming increasingly popular. An important place in the global economy is occupied by oilseeds, in particular rapeseed, which production volumes are increasing in the Russian Federation every year. Rapeseed (Brassicaceae napus) is of great interest due to its high oil content (39.80–46.00%) and rich fatty acid composition, while cake and meal formed in the process of oil production are characterized by a significant content of crude protein (35.00–45.00%) and crude fiber (8.20–17.50%); however, they are used mainly as a feed additive. Recent studies on the processing of rapeseed waste indicate the value of this raw material as a source of dietary protein, which has a balanced amino acid profile and a high degree of digestibility (up to 85%). To obtain protein, rapeseed processing is envisaged: cleaning, grinding, cold pressing at a temperature of ≤ 40 °C, fat extraction with a solvent. At the next stages, the protein is extracted with 0.1–0.5 M NaCl at pH 5.3–12.0 and a temperature of 5–30 °C for 1 hour. The extracted protein is precipitated at the isoelectric point (pH 4.0) with HCl, separated from the mixture and neutralized. The result is a protein isolate with a protein content of 90.0–98.7%. It is possible to increase the quality and yield of the protein product due to the additional stage of processing the defatted cake with cellulolytic enzyme preparations. In this case, additional studies are required to determine the substrate specificity of commercial cellulase enzyme preparations and the optimal hydrolysis conditions. The parameters of extraction and precipitation of the protein in the case of using the stage of enzymatic lysis should also be specified.

Вплив параметрів ферментативної обробки на вихід пресової гарбузової олії та її властивості

The influence of parameters of pumpkin seeds pretreatment with enzyme preparations of proteolytic and cellulolytic activity on the yield of press oil, its composition and quality parameters were studied in this work. The enzyme preparation PENICILOPEPSIN (Enzym Biotech, Ukraine) with proteolytic activity and CELLULAD (Enzym Biotech, Ukraine) with cellulolytic, hemicellulase and xylanase activity were used for pretreatment. Enzymatic treatment of seeds was carried out with a mixture of these preparations at a ratio of 7:3 for 2 h at a pH 5.2 and 48—54°C. The following parameters of enzymatic pretreatment were studied: the amount of the enzyme mixture, which varied from 0.3% to 2.4% of the weight of the seeds and the amount of moisture added together with the enzymes — from 15% to 50%. It was established that the rational parameters of the enzymatic treatment of pumpkin seeds are the mass of the enzyme mixture of 0.6% and the amount of moisture added with enzymes — 35% of the seed mass. Under such technological parameters, the yield of press pumpkin oil was 65.6%, which is almost 6% higher than in the control sample. The acid and anisidine value of the control oil sample and the oil extracted after the enzymatic treatment of the seeds did not differ significantly. However, the content of peroxide compounds in the oil extracted after enzymatic treatment of seeds was lower. The total antioxidant activity of the oil, determined by the reaction of the 2,2-diphenyl-1- picrylhydrazyl radicals quenching within 30 min, was higher in the oil samples after seed pretreatment at the maximum tested moisture (50%). The obtained results indicate that the enzymatic pretreatment of pumpkin seeds with proteolytic and cellulolytic enzyme preparations is a promising method of oil yield increasing.

Optimization of the use of cellulolytic enzyme preparation for the extraction of health promoting anthocyanins from black carrot using response surface methodology

Anthocyanin-rich extracts from black carrots are being considered as a candidate replacer for the red colour in processed foods. The present investigation optimizes the extraction of anthocyanins with high phenolic content and low degradation parameters from black carrots using a cellulolytic multi-enzyme preparation known as Viscozyme. The optimized conditions for Viscozyme using a Box-Behnken design (BBD) of response surface methodology (RSM) were as follow: temperature (50.2 °C), extraction time (58.4 min) and enzyme concentration (0.20%). The predicted value of anthocyanins content was 1380 mg/L, which was near to the optimized experimental value of 1375 mg/L. The extracted anthocyanins based on above mentioned conditions exhibited the lowest degradation parameters such as degradation index (DI) of (0.86), browning index of (BI) (1.31) and were characterized with cyanidin 3-sinapoylxylosylglucosylgalactoside as being the most abundant. The findings clearly reveal that Viscozyme-assisted extraction (VAE) is the best approach for extracting superior quality extracts from black carrots with high anthocyanin and other phenolic component concentrations.

Разработка инновационного способа получения биологически активных соединений пивной дробины

Introduction. Brewery mash, or brewer’s spent grain (BSG), is a by-product of brewing industry. It is known to contain valuable biologically active substances. However, their extraction is complicated by the presence of various polymers. The research featured various physicochemical methods for obtaining valuable biological compounds from brewery waste. The new method modified complex non-starch polysaccharides, lignin, arabinoxylans, and other high-molecular compounds associated with phenolic compounds. The research objective was to solve the problem of recycling industrial by-products that accumulate in large quantities and require expensive processing or disposal. The paper introduces new technological approaches for deep processing of BSG as a source of secondary raw materials in order to obtain extracts fortified with polyphenolic compounds.
 Study objects and methods. The research featured BSG from malt subjected to treatment with ECA-activated water (catholyte with pH 9.6 ± 0.1), followed by enzymatic hydrolysis of cellulolytic enzyme preparations and extraction with a polar solvent of the resulting free polyphenolic substances. The experiments were based on standard methods for assessing the content of various biologically active substances.
 Results and discussion. A 70% water-ethanol solution proved to be optimal at the BSG:extractant ratio of 2:1, process temperature = 50 ± 2°C, and extraction time = 60 ± 5 min. Under the same conditions, 70 %vol. of beer distillate made it possible to extract phenolic acids, flavonoid rutin, irreplaceable and nonessential amino acids, and non-starch polysaccharide β-glucan from the BSG matrix. The BSG treatment with 1M NaOH solution delivered viscous hydrolysates fortified with flavonoids rutin and quercetin, which did not happen when acid hydrolysis was used. The combined use of ECA-treated water (catholyte with pH 9.6 ± 0.1) for 24 ± 0.05 h, combined with biocatalysis with the enzyme preparation Viskoflo MG for 2 ± 0.05 h, made it possible to obtain BSG extracts with a high content of phenolic acids and aldehydes, as well as flavonoid rutin.
 Conclusion. The study revealed the mechanism of hydrolytic decomposition of BSG non-starch polysaccharides, considering the compounds contained in the extracts. The BSG hydrolysates fortified with various phenolic compounds can be used in various food technologies, e.g., in fermented drinks.

Bioprocessing of wheat to produce food and technical products

The article presents the parameters and conditions for the formation of gluten in the laboratory. The influence of some technological parameters in the preparation of water-powdery suspension on the yield of gluten was studied. It was found that for the formation of a strong gluten skeleton, the temperature must be 50 ° C, the duration of the batch - 30 minutes, and the hydromodule - 1.5:1. It is under these conditions that the binding of the gluten and gliadin protein fractions in the gluten frame. It is proved that due to cellulolytic enzyme preparations, sequential hydrolysis of non-starch polysaccharides to glucose is carried out, as well as dispersion of starch and other high-molecular substances in the grain composition. This increases the quantity of fermentable substances and increases the quantity of alcohol produced. Processing of wort obtained after extraction of gluten from water-flour suspension results in higher ethanol concentration, glucose fermentation rate, and ethanol production rate. In addition, gluten can be used as a value-added by-product in a wheat-based bioprocessing plant, which significantly increases the cost-effectiveness of the process.

Analysis of the chemical composition and properties of the main and side products during the implementation of new technologies for producing ethanol

The article studied yeast composition isolated from the separation of the mash, bran separated in the grinding step, as well as a protein supplement obtained by mixing these two products. Crude fat content ranges from 4,7 to 6,2%, crude protein content is 38%, which is 20% lower than in the fodder yeast, but 14% higher than that of the bran. It is also found that the resulting protein product enriched in micro- and macro-elements, as well as readily hydrolyzable and water soluble carbohydrates. crude fiber content is 5,5%, which is almost 2,5 times lower than in the bran. The composition of the feed protein supplement obtained with the introduction at the stage of aqueous suspension of wheat powdery cellulolytic enzyme preparation Viskolaza 150 L and without it. The enzyme preparation was added in the step of preparing aqueous suspensions and wheat powdery dosage of 0,01% by weight of the feedstock. protein feed additives obtained with and without addition of cellulolytic enzyme preparation of high quality crude protein content and protein. Thus, the content of crude protein in the protein with the introduction of an additive in an aqueous suspension of powdery Viskolazy 150 L was 37% whereas 34% without the enzyme preparation. The amount of soluble polysaccharides and the mass fraction of ash were practically at the same level of from 2,4 to 5% and from 5,5 to 7,0%, respectively. fiber content of the protein supplement using Viskolazy 150 L was 4,2-6,1%, which is 2,5 times lower than in the protein supplement obtained without enzyme preparation.

Evaluation of Cellulolytic Enzyme-Assisted Microwave Extraction of Punica granatum Peel Phenolics and Antioxidant Activity.

Conventional techniques for phenolics extraction from pomegranate (Punica granatum) peel (PP) have several insufficiencies like longer time duration, bioactive degradation, excess use of harmful chemicals and solvents. In the present study, we established the synergistic use of two non-conventional extraction strategies i.e., enzyme-assisted extraction (EAE) using a cellulolytic enzyme preparation (Viscozyme) followed by microwave-assisted extraction (MAE) for efficient recovery of phenolics from PP. This optimized method was individually compared with EAE, MAE, and conventional solvent extraction (CSE) methods for recovering PP phenolics with maximum antioxidant activity (AOA). Extracts were analyzed for AOA using ferric reducing antioxidant power (FRAP) and cupric reducing AOA (CUPRAC) methods. Response surface methodology (RSM), was used as an optimization tool to achieve maximum yield of phenolics and with highest AOA at power 443.5W, time 131.0min, and solvent-to-solid ratio 23.6:1. The predicted values for maximum phenolics and AOA obtained through RSM were 305mg GAE/g, 1788μmol TE/g (FRAP) and 2641μmol TE/g (CUPRAC), respectively. Phenolic contents of only 94.6, 165.46, and 197.6mg GAE/g were achieved through CSE, EAE and MAE, respectively. Here we substantiate the auxiliary role of Viscozyme and microwave treatment in achieving high phenolic content and AOA from PPs. Phenolic rich extracts are known to act as multi-target ligands that inhibit various enzymes involved in diseases like Alzheimer's, Parkinson's, and diabetes mellitus. The extract can be commercially exploited for the development of functional foods, supplements, and natural preservatives.

Demonstration‐scale enzymatic saccharification of sulfite‐pulped spruce with addition of hydrogen peroxide for LPMO activation

AbstractThe saccharification of lignocellulosic materials like Norway spruce is challenging due to the recalcitrant nature of the biomass, and it requires optimized and efficient pretreatment and enzymatic hydrolysis processes to make it industrially feasible. In this study, we report successful enzymatic saccharification of sulfite‐pulped spruce (Borregaard's BALI™ process) at demonstration scale, achieved through the controlled delivery of hydrogen peroxide (H2O2) for the activation of lytic polysaccharide monooxygenases (LPMOs) present in the cellulolytic enzyme preparation. We achieved 85% saccharification yield in 4 days using industrially relevant conditions – that is, an enzyme dose of 4% (w/w dry matter of substrate) of the commercial cellulase cocktail Cellic CTec3 and a substrate loading of 12% (w/w). Addition of H2O2 and the resulting controlled and high LPMO activity had a positive effect on the rate of saccharification and the final sugar titer. Clearly, the high LPMO activity was dependent on feeding the reactors with the LPMO co‐substrate H2O2, as in situ generation of H2O2 from molecular oxygen was limited. These demonstration‐scale experiments provide a solid basis for the use of H2O2 to improve enzymatic saccharification of lignocellulosic biomass at large industrial scale.© 2020 The Authors. Biofuels, Bioproducts, and Biorefining published by Society of Chemical Industry and John Wiley & Sons, Ltd.

BIOCONVERSION OF SECONDARY PRODUCTS OF PROCESSING OF GRAIN CEREALS CROPS

A method has been developed for the production of organic ingredients from secondary products resulting from the high-quality grinding of triticale and wheat into flour, which involves the enzymatic action of amylolytic enzymes to release starch polysaccharides while preserving the native properties of dietary fibers and biologically active substances associated with them. To a large extent, the features of the properties of the obtained ingredients are due to the number and composition of the components of dietary fiber of grain, as well as the morphological features of their structure. It is shown that the viscosity of aqueous colloidal systems at a concentration of soluble dietary fiber of the ingredient 0.5 % increases 11 times; at a concentration of 1.0 % — 30 times, forming a viscous gel-like structure. This allows them to be used for gelling, thickening and stabilization of aquatic food systems. The use of ingredients with a high content of NLP in baking is possible only taking into account their water absorption capacity. A method for the enzymatic modification of secondary products of processing of grain triticale was developed. On the basis of the study of the kinetics and efficiency of the effect of proteolytic and cellulolytic enzyme preparations (EP) and their compositions, optimal conditions for enzymatic modification (the EP dosage is 0.5…0.75 units of PA/g of bran, 0.3…0.4 units of CA/g of bran, the optimum temperature is 40–50 °С, pH is 5.0 and 3.5, the duration of reactions is 1.5 hours) have been determined. The use of cellulolytic EP allowed to increase the amount of reducing substances and soluble protein by 1.5–2.5 times in comparison with the control sample. The biomodified bran obtained using the MEC «Shearzyme 500 L» + «Neutrase 1.5 MG» and «Viscoferm L» + «Distizym Protacid Extra» has a high degree of hydrolysis of non-starch polysaccharides and proteins, is characterized by a certain ratio of high-, medium-, low-molecular peptides and amino acids, has different functional and technological properties. They can be used in the production of a wide range of general-purpose, functional and treatmentand-prophylactic food products.

Intensification of grain sprouting process using complex enzyme preparations

The regimes were grounded for enzymatic treatment of food wheat and peas grain to intensify seedling production. Optimization of grain germination process envisages application at the steeping stage of cellulolytic enzyme preparations (cellulase, xylanase, β-glucanase) and amylolytic enzymes in 0.005 % concentrations during 3 h. The highest efficiency among tested biopreparations for wheat sprouting was shown by Viscoferm (Novozymes, Denmark) and Compliferm (Institute of Microbiology, NAS Belarus), whereas Viscoferm proved the most active in terms of peas germination. Enzymatic treatment results in reduced duration of wheat grain sprouting by 2–4 h, pea seed sprouting by 4–5 h on the average, and increased seedlings yield, raising thereby profitability of the process.

Lignin from hydrothermally pretreated grass biomass retards enzymatic cellulose degradation by acting as a physical barrier rather than by inducing nonproductive adsorption of enzymes

BackgroundLignin is known to hinder efficient enzymatic conversion of lignocellulose in biorefining processes. In particular, nonproductive adsorption of cellulases onto lignin is considered a key mechanism to explain how lignin retards enzymatic cellulose conversion in extended reactions.ResultsLignin-rich residues (LRRs) were prepared via extensive enzymatic cellulose degradation of corn stover (Zea mays subsp. mays L.), Miscanthus × giganteus stalks (MS) and wheat straw (Triticum aestivum L.) (WS) samples that each had been hydrothermally pretreated at three severity factors (log R0) of 3.65, 3.83 and 3.97. The LRRs had different residual carbohydrate levels—the highest in MS; the lowest in WS. The residual carbohydrate was not traceable at the surface of the LRRs particles by ATR-FTIR analysis. The chemical properties of the lignin in the LRRs varied across the three types of biomass, but monolignols composition was not affected by the severity factor. When pure cellulose was added to a mixture of LRRs and a commercial cellulolytic enzyme preparation, the rate and extent of glucose release were unaffected by the presence of LRRs regardless of biomass type and severity factor, despite adsorption of the enzymes to the LRRs. Since the surface of the LRRs particles were covered by lignin, the data suggest that the retardation of enzymatic cellulose degradation during extended reaction on lignocellulosic substrates is due to physical blockage of the access of enzymes to the cellulose caused by the gradual accumulation of lignin at the surface of the biomass particles rather than by nonproductive enzyme adsorption.ConclusionsThe study suggests that lignin from hydrothermally pretreated grass biomass retards enzymatic cellulose degradation by acting as a physical barrier blocking the access of enzymes to cellulose rather than by inducing retardation through nonproductive adsorption of enzymes.

High activity CAZyme cassette for improving biomass degradation in thermophiles

BackgroundThermophilic microorganisms and their enzymes offer several advantages for industrial application over their mesophilic counterparts. For example, a hyperthermophilic anaerobe, Caldicellulosiruptor bescii, was recently isolated from hot springs in Kamchatka, Siberia, and shown to have very high cellulolytic activity. Additionally, it is one of a few microorganisms being considered as viable candidates for consolidated bioprocessing applications. Moreover, C. bescii is capable of deconstructing plant biomass without enzymatic or chemical pretreatment. This ability is accomplished by the production and secretion of free, multi-modular and multi-functional enzymes, one of which, CbCel9A/Cel48A also known as CelA, is able to outperform enzymes found in commercial enzyme preparations. Furthermore, the complete C. bescii exoproteome is extremely thermostable and highly active at elevated temperatures, unlike commercial fungal cellulases. Therefore, understanding the functional diversity of enzymes in the C. bescii exoproteome and how inter-molecular synergy between them confers C. bescii with its high cellulolytic activity is an important endeavor to enable the production of more efficient biomass degrading enzyme formulations and in turn, better cellulolytic industrial microorganisms.ResultsTo advance the understanding of the C. bescii exoproteome we have expressed, purified, and tested four of the primary enzymes found in the exoproteome and we have found that the combination of three or four of the most highly expressed enzymes exhibit synergistic activity. We also demonstrated that discrete combinations of these enzymes mimic and even improve upon the activity of the whole C. bescii exoproteome, even though some of the enzymes lack significant activity on their own.ConclusionsWe have demonstrated that it is possible to replicate the cellulolytic activity of the native C. bescii exoproteome utilizing a minimal gene set, and that these minimal gene sets are more active than the whole exoproteome. In the future, this may lead to more simplified and efficient cellulolytic enzyme preparations or yield improvements when these enzymes are expressed in microorganisms engineered for consolidated bioprocessing.

Using an Inducible Promoter of the Glucoamylase Gene to Construct New Multienzyme Complexes from Penicillium verruculosum

New recombinant strains of Penicillium verruculosum are created using a new plasmid construct based on an inducible promoter of glucoamylase gene (gla1) that secretes heterologous xylanase E (XylE) from P. canescens. New biocatalysts are produced that contain cellulolytic enzyme preparations (EPs) enriched with XylE. The amount of XylE in the recombinant EPs varies in the optimum range of 11–24% of the total protein while generally preserving the P. verruculosum cellulose complex. The hydrolytic activity of the new EPs with respect to polymeric plant-derived substrates exceeds that of EPs produced using other expression systems and commercial preparations. The new EP glaX-17 surpasses in particular a control EP based on the recipient strain by 13% in the efficiency of hydrolyzing aspen wood and is 20% more efficient than the commercial EP Accelerase Duet. The new EP glaX-17 displays 25% greater efficiency (35–43%) during the hydrolysis of wheat bran than the commercial EP Accelease Duet. The effectiveness is demonstrated of using the new gla1 promoter for the production of EPs (biocatalysts) while preserving the balanced cellulose complex of the strain and optimum yield of heterologous XylE required for the deep hydrolysis of xylan-containing plant biomass.

Chokeberry pomace valorization into food ingredients by enzyme-assisted extraction: Process optimization and product characterization

Currently berry juice pressing residues are used inefficiently due to a lack of innovative processes of their valorization. This study reports valorization of chokeberry pomace via combined high pressure fractionation and enzyme-assisted extraction (EAE), utilizing commercially available cellulolytic and xylanolytic enzyme preparations, namely Viscozyme L and CeluStar XL. The results obtained indicate that optimized enzyme treatment (E/S 6% v/w, 40°C, pH 3.5, 7h), especially using Viscozyme L, facilitates pomace cell-wall breakdown and: (1) increases the yield of water-soluble fraction by 44–113%, monosaccharide content by 12–140%, total phenolic content by 29–41%, radical scavenging capacity by 20–39%; (2) promotes some alterations in phenolic acid and flavonol profiles of EAE-derived supernatants. For the first time antioxidant potential of solid residues from enzyme-treated berry pomaces was evaluated too, indicating that all fractions could be utilized as a low-cost source of high added value functional ingredients with various scientific and commercial applications.

Synergies in coupled hydrolysis and fermentation of cellulose using a Trichoderma reesei enzyme preparation and a recombinant Saccharomyces cerevisiae strain.

We describe a procedure by which filter paper is digested with a cellulolytic enzyme preparation, obtained from Trichoderma reesei cultivated under solid state fermentation conditions and then fermented by a recombinant Saccharomyces cerevisiae strain. The yeast strain produces a β-glucosidase encoded by the BGL1 gene from Saccharomycopsis fibuligera that quantitatively and qualitatively complements the limitations that the Trichoderma enzyme complex shows for this particular activity. The supplemental β-glucosidase activity fuels the progression of cellulose hydrolysis and fermentation by decreasing the inhibitory effects caused by the accumulation of cellobiose and glucose. Fermentation of filter paper by this procedure yields ethanol concentrations above 70g/L.

Enzymatic Activity of Some Industrially-Applied Cellulolytic Enzyme Preparations

Abstract Enzymatic hydrolysis is the essential step in the production of 2nd generation biofuels made from lignocellulosic biomass, i.e. agricultural or forestry solid wastes. The enzyme-catalysed robust degradation of cellulose and hemicellulose to monosaccharides requires the synergistic action of the independent types of highly-specific enzymes, usually offered as ready-to-use preparations. The basic aim of the study was to experimentally determine the enzymatic activity of two widely industrially-applied, commercially available cellulolytic enzyme preparations: (i) Cellic® CTec2 and (ii) the mixture of Celluclast® 1.5L and Novozyme 188, in the hydrolysis of pre-treated lignocellulosic biomass, i.e. (a) energetic willow and (b) rye straw, or untreated (c) cellulose paper as well, used as feedstocks. Before the hydrolysis, every kind of utilized lignocellulosic biomass was subjected to alkaline-based (10% NaOH) pre-treatment at high-temperature (121°C) and overpressure (0.1 MPa) conditions. The influence of the type of applied enzymes, as well as their concentration, on the effectiveness of hydrolysis was quantitatively evaluated, and finally the enzyme activities were determined for each of tested cellulolytic enzyme preparations.

Fractionation of sea buckthorn pomace and seeds into valuable components by using high pressure and enzyme-assisted extraction methods

Sea buckthorn pomace and seeds were processed via 3-step fractionation using supercritical carbon dioxide (SFE-CO2), pressurized ethanol (PLE-EtOH) and enzyme-assisted (EAE) extraction. SFE-CO2 yielded 146 and 135 g/kg of lipophilic fraction from pomace and seeds, respectively. PLE-EtOH of SFE-CO2 residues recovered 158 g/kg of polar constituents from the pomace and 5-fold lower amount from seeds. Finally, the treatment of PLE-EtOH residues with cellulolytic and xylanolytic enzyme (Viscozyme, CeluStar XL) preparations increased the amount of soluble constituents by 24–80%, as compared to enzyme-untreated samples. CO2 extracts of pomace and seeds contained 630 and 2223 mg/kg tocopherols, respectively; while PLE-EtOH extracts contained sugar acids, phenolics and triterpenes. EAE increased glucose, fructose and maltose content. The total phenolic content (2–22 g gallic acid/kg DW prior SFE-CO2) and radical scavenging capacity (6–166 g Trolox/kg DW) indicate that both soluble and insoluble EAE fractions could be utilized as a low-cost source of functional ingredients for their potential applications in foods.

The Application of the Inducible Promoter of Glucoamylase Gene for Preparation of New Multienzyme Complexes Penicillium verruculosum

A new plasmid construction based on the inducible promoter of glucoamylase gene ( gla1 ) was used for the obtaining of new recombinant strains Penicillium verruculosum that secrete heterological xylanase E (XylE) P. canescens . New biocatalysts were developed which are XylEenriched cellulolytic enzyme preparations (EP). The content of XylE in recombinant EP varied within the optimal range from 11 to 24 % of the general protein pool at the preservation (in total) of the cellulase complex P. verruculosum . In comparison to EP prepared using the other expression systems and commercial preparations, the new EP demonstrated a higher hydrolytic activity with respect to polymers of plant substrates. For example, the efficiency of new EP glaX-17 to hydrolysis of aspenwood was higher by 13 % than that of the control EP agent and by 20 % than that of the commercial EP Accelerase Duet. In hydrolysis of wheat bran, glaX-17 was 35–43 % and 25 % more efficient than the control EP and the commercial EP Accelerase Duet, respectively. The efficiency of the new gla1 promoter for the production of EP (biocatalysts) at the preserved balanced cellulase complex of the strain and optimal yield of heterological XylE (that are necessary for complete hydrolysis of xylan-containing plant biomass) was demonstrated.

Evaluation of efficiency of using the enzyme preparation «Celloviridin Dx20» in the content of pig feed

One of the main problems in the feeding of farm animals is the low degree of digestion and assimilation of nutrients in the feed used in animal production. In the bodies of animals such as pigs and poultry enzymes and microflora which are necessary for breaking down cellulose and other polysaccharides of vegetable raw materials are absent, therefore they not only fail to be digested in the gastrointestinal tract, but also obstruct access of other digestive enzymes to the other feed components, in particular intracellular proteins. The only way to solve this problem is deliberate introduction of enzymes into food. Depending on the type and composition of the feed, amylolytic, proteolytic, cellulolytic enzyme preparations are used. The purpose of this work is to assess the effect of the enzyme preparation "Celloviridin Dx20" as a fodder component on the growth and biochemical parameters of young pigs. For this purpose, a scientific and economic experiment on feeding young pigs was conducted at the agro-industrial farm«Niva» (Novonikolaevka village, Dnipropetrovsk region), in which the enzyme preparation «Celloviridin Dx20», which promotes the breakdown of plant cell polysaccharides (cellulose, xylans) was introduced to a standard feed mixture used in swine husbandry. The composition of the ration consisted of 60% barley, 20% wheat and 20% of the protein-vitamin-mineral complex «ShenPigStart». The results of the experiment showed that the introduction into the feed mixture of the enzyme preparation at 100 g per ton helps to increase the body weight of pigs by 11.6% compared to the control, can increase the digestion rate of fibre (13.8%), protein and fat. These results can be explained by the mechanism of action of the enzyme complex, which consists in the hydrolysis of polysaccharides of plant cell walls in the feed in the digestive tract of pigs, the formation of digestable polysaccharides and the release of additional quantities of intracellular proteins. The result of these phenomena is also a positive trend in feed nitrogen assimilation in animals of the experimental group: a 19% decrease in the amount of physiological secretions and a 12.5% increase in nitrogen use. At the same time hematology (hemoglobin, erythrocytes, leukocytes), calcium and phosphorus metabolism remained unchanged. This shows that the preparation used does not accumulate in the body and gradually loses enzymatic activity, and under the influence of the pigs’ stomach proteases it is broken down into separate amino acids which are absorbed by the animals. The resulting stimulatory effect of using "Celloviridin Dх20" has not only physiological but also economic importance, as it helps reduce the time for the pigs to reach slaughter weight (close to 100 kg) by 11 days, and consequently, reduces feed intake and its costs in rearing pigs on farms. Therefore, it is advisable to use "Celloviridin Dх20" in swine husbandry.

Composition and microstructure alteration of triticale grain surface after processing by enzymes of cellulase complex

It is found that the pericarp tissue of grain have considerable strength and stiffness, that has an adverse effect on quality of whole-grain bread. Thereby, there exists the need for preliminary chemical and biochemical processing of durable cell walls before industrial use. Increasingly used in the production of bread finds an artificial hybrid of the traditional grain crops of wheat and rye - triticale, grain which has high nutritional value. The purpose of this research was to evaluate the influence of cellulose complex (Penicillium canescens) enzymes on composition and microstructure alteration of triticale grain surface, for grain used in baking. Triticale grain was processed by cellulolytic enzyme preparations with different composition (producer is Penicillium canescens). During experiment it is found that triticale grain processing by enzymes of cellulase complex leads to an increase in the content of water-soluble pentosans by 36.3 - 39.2%. The total amount of low molecular sugars increased by 3.8 - 10.5 %. Studies show that under the influence of enzymes the microstructure of the triticale grain surface is changing. Microphotographs characterizing grain surface structure alteration in dynamic (every 2 hours) during 10 hours of substrate hydrolysis are shown. It is found that the depth and direction of destruction process for non-starch polysaccharides of grain integument are determined by the composition of the enzyme complex preparation and duration of exposure. It is found, that xylanase involved in the modification of hemicelluloses fiber having both longitudinal and radial orientation. Hydrolysis of non-starch polysaccharides from grain shells led to increase of antioxidant activity. Ferulic acid was identified in alcoholic extract of triticale grain after enzymatic hydrolysis under the influence of complex preparation containing cellulase, xylanase and β-glucanase. Grain processing by independent enzymes containing in complex preparation (xylanase and β-glucanase) shows that more significant role in polysaccharide complex composition and grain surface microstructure alteration belongs to xylanase. Grain processing by independent of cellulolytic enzymes may decrease the strength of pericarp tissue of grain and improved sensory characteristics of the bread.