The use of dietary fibre to optimize microbial gut function in pigs, with particular consideration of dietary cereal grains and legumes

This chapter examines interactions of dietary fibre components of pig diets with GIT microbiota, emphasizing cereals and legumes fed to pigs. Carbohydrate composition of these feedstuffs are described, and their relationship to metabolic activity of the porcine intestinal microbiota and interactions with the host. Fermentable carbohydrates which act as substrates for microbial metabolism are described, followed by an assessment of cereals and legumes as potential modulators of intestinal microbiota. Past work focussed on purified extracts, but attention is now focussing on whole grains or their fractions such as brans, in terms of effects on microbial populations. Such studies are showing the positive consequences of mixtures of DF in the form of complex plant cellular structures, rather than single refined ingredients, to achieve beneficial health outcomes. Further work is also needed to define appropriate quantities and types of DF to achieve desired effects whilst minimising negative outcomes.

Hydrolysis of S. platensis Using Sulfuric Acid for Ethanol Production

S. platensis is a microalga that contains carbohydrate composition of 30.21% which makes it potential to be used as raw material for ethanol production. Hydrolysis of S. platensis is the first step for converting its carbohydrates into monosaccharides. The second step is fermentation of monosaccharides into ethanol. This research aims to study the effect of temperature and microalgae concentration on the hydrolysis of S. platensis using sulfuric acid as catalyst. This research was conducted using 300 mL sulfuric acid of 2 mol/L, hydrolysis temperatures of 70, 80 and 90 °C, and microalgae concentrations of 20, 26.7, and 33.3 g/L. The effect of temperature is significant in the hydrolysis of S. platensis using sulfuric acid. At microalgae concentration of 20 g/L and hydrolysis time of 35 minutes, the higher the temperatures (70, 80, and 90 °C), the more the glucose yields would be (8.9, 13.5, and 22.9%). This temperature effect got stronger when the hydrolysis was running for 15 minutes. Every time the hydrolysis temperature increased by 10 °C, the glucose yield increased by 13.0% at microalgae concentration of 33.3 g/L. At temperature of 90 °C and time of 35 minutes, the higher the microalgae concentrations (20, 26.7, and 33.3 g/L), the higher the glucose yields would be (25.5, 27.7, and 28.2%). The highest glucose concentration obtained was 2.82 g/L at microalgae concentration of 33.3 g/L, temperature of 90 °C, and time of 35 minutes.

The Action of Recombinant Human Lysosomal α-Glucosidase (rhGAA) on Human Liver Glycogen: Pathway to Complete Degradation

Glycogen is present in all tissues, but it is primarily stored in the liver and in muscle. As a branched chain carbohydrate, it is broken down by phosphorylase and debrancher enzymes, which are cytoplasmic. It is also degraded by a lysosomal α-glucosidase (GAA) also known as acid α-glucosidase and lysosomal acid α-glucosidase. The deficiency of GAA in patients is known as Pompe disease, and the phenotypes as infantile, juvenile and later onset forms. Pompe disease is treated by enzyme replacement therapy (ERT) with a recombinant form of rhGAA. Following ERT in Pompe mice and human patients there is residual carbohydrate material present in the cytoplasm of cells. The goal of this work is to improve ERT and attempt to identify and treat the residual cytoplasmic carbohydrate. Initial experiments were to determine if rhGAA can completely degrade glycogen. The enzyme cannot completely degrade glycogen. There is a residual glycosylated protein as well as a soluble glycosylated protein, which is a terminal degradation product of glycogen and as such serves as a biomarker for lysosomal glycogen degradation. The glycosylated protein has a very unusual carbohydrate composition for a glycosylated protein: m-inositol, s-inositol and sorbitol as the major carbohydrates, as well as mannitol, mannose, glucose and galactose. This work describes the residual material which likely contains the same protein as the soluble glycosylated protein. The biomarker is present in serum of control and Pompe patients on ERT, but it is not present in the serum of Pompe mice not on ERT. Pompe mice not on ERT have another glycosylated protein in their serum which may be a biomarker for Pompe disease. This protein has multiple glycosylation sites, each with different carbohydrate components. These glycosylated proteins as well as the complexity of glycogen structure are discussed, as well as future directions to try to improve the outcome of ERT for Pompe patients by being able to monitor the efficacy of ERT in the short term and possibly to adjust the timing and dose of enzyme infusions.

Preparation of maltotetraose starch molasses using Maltotetraohydrolase Optimalt 4G

Одним из направлений решения актуальной задачи по расширению ассортимента сахаристых крахмалопродуктов является разработка способов получения мальтоолигосахаридов с различным углеводным составом, имеющих большой потенциал для использования в качестве пищевых добавок и ингредиентов продуктов здорового питания. Целью работы было изучение возможности получения нового вида крахмальной патоки с высоким содержанием мальтотетраозы с применением препарата мальтотетраогидролазы Optimalt 4G. Объектами являлись кукурузный крахмал, препараты альфа-амилазы Liquazyme Supra 2.8X и пуллуланазы Promozyme D6 (Novozymes), мальтотетраогидролазы Optimalt 4G (Du Pont), а также продукты гидролиза крахмала. Использованы методы определения массовой доли редуцирующих веществ (РВ), сухого вещества (СВ), углеводного состава, динамической вязкости гидролизатов и другие стандартные методы анализа. При осахаривании частично гидролизованной суспензии крахмала с концентрацией СВ 33 % и содержании РВ 7,9 % мальтотетраогидролаза наиболее активно действует в первые 3 ч осахаривания с резким снижением вязкости гидролизата с 197,6 до 20,0 мПа·с. При этом исчезали практически все олигосахариды с DР5-DР10, массовая доля ВМС снизилась на 60 %, увеличилось количество низкомолекулярных сахаров, а массовая доля мальтотетраозы составила 44,81 % против 2,38 % в исходном субстрате. Максимальное содержание мальтотетраозы 49,8 % в данных условиях наблюдалось в гидролизате после 8 ч инкубации, далее оно медленно снижалась. Установлена возможность получения крахмальной патоки с высоким содержанием мальтотетраозы до 45-48 % с использованием в качестве катализаторов гидролиза крахмала вышеуказанных ферментных препаратов. Использование композиции мальтотетраогидролазы с пуллуланазой увеличивает содержание мальтотетраозы на 1,5-2 %, снижает вязкость гидролизатов и позволяет сократить продолжительность гидролиза. Полученные данные являются актуальной основой для оптимизации технологических параметров процесса осахаривания с применением мальтотетраогидролазы с целью разработки технологии получения нового вида крахмальной патоки. One of the directions of solving the current problem of expanding the range of sugar starch products is the development of methods for producing maltooligosaccharides with a different carbohydrate composition, which have great potential for use as food additives and ingredients of healthy foods. The aim of the work was to study the possibility of obtaining a new type of starch syrup with a high content of maltotetraose using the preparation Maltotetraohydrolase Optimalt 4G. The objects were: corn starch, alpha-amylase preparations Liquazyme Supra 2.8X and pullulanase Promozyme D6 (Novozymes), maltotetraohydrolase Optimalt 4G (Du Pont), as well as starch hydrolysis products. Methods were used to determine the weight fraction of reducing substances (RS), dry solid (DS), carbohydrate composition, dynamic viscosity of hydrolysates and other standard methods of analysis. When a partially hydrolyzed suspension of starch with a concentration of DS 33 % and RS 7.9 % is saccharified, maltotetraohydrolase is most active in the first 3 hours of saccharification with a sharp decrease in the viscosity of the hydrolysate from 197.6 to 20.0 mPa·s. At the same time, almost all oligosaccharides with DP5-DP10 disappeared, the weight fraction of WMS decreased by 60 %, the number of low-molecular sugars of DP1-3 increased, and the weight fraction of maltotetraose was 44.81 % versus 2.38 % in the original substrate. The maximum maltotetraose content of 49.8 % under these conditions was observed in the hydrolysate after 8 hours of incubation, then it slowly decreased. It has been found that in order to obtain hydrolysates with a low DP1-3 content and increased (more than 45 %) maltotetraose, the degree of preliminary hydrolysis must be maintained within the range of RS 8-15 % by DS at an enzyme dosage of 0.06-0.08 units WMC/g DS starch and duration of saccharification up to 20 h. Use of maltotetraohydrolase composition with pullulanase during saccharification increases degree of hydrolysis and content of maltotetraose by 1.5-2 %, reduces viscosity of hydrolysates and reduces duration of hydrolysis. It is possible to produce starch syrup with high content of maltotetraose (up to 45-48 %) using the above enzyme preparations. The obtained data are the current basis for optimization of technological parameters of saccharification process with application of maltotetraohydrolase in order to develop technology of production of new kind of starch syrup.

Modeling the functional properties of minced meat semi-finished product with a protein-carbohydrate composition of a combined composition

Современными исследованиями доказано, что большинство пищевых продуктов, в том числе полуфабрикатов, используемых на предприятиях общественного питания, значительно обеднены полезными нутриентами. В настоящее время большое внимание уделяется технологиям обогащения продукции биологически активными веществами и функциональными ингредиентами, вводимыми в регулярно употребляемые населением пищевые продукты в количествах, соответствующих физиологическим потребностям человека. В последние годы наблюдается активное внедрение цифровых информационных технологий в проектирование многокомпонентных продуктов питания. При помощи математических систем можно осуществлять оптимизацию рецептурного состава многокомпонентных пищевых систем, проводить интегральную оценку сбалансированности проектируемых продуктов питания. Целью исследования является изучение нутриентного состава и оптимизация рецептуры мясного фарша с наполнителем в виде белково-углеводной композиции комбинированного состава посредством проектирования, регламентирующего этапы создания пищевых продуктов с заданными функциональными свойствами. При этом были исследованы химический, аминокислотный и жирнокислотный состав опытных образцов мясных фаршей с наполнителем различного процентного содержания в сравнении с контрольным образцом. В данной статье приведены результаты исследований мясного фарша с заменой мясного сырья наполнителем в количестве 10-25%. В итоге была определена оптимальная рецептура сбалансированного по составу мясного фарша с белково-углеводной композицией в количестве 10%. Была разработана математическая модель мясного полуфабриката с повышенной пищевой и биологической ценностью. Modern research has proven that most food products, including semi-finished products used in public catering establishments, are significantly depleted in useful nutrients. At present, much attention is paid to technologies for enriching products with biologically active substances and functional ingredients introduced into food products regularly consumed by the population in quantities corresponding to the physiological needs of a person. In recent years, there has been an active introduction of digital information technologies in the design of multicomponent food products. With the help of mathematical systems, it is possible to optimize the recipe composition of multicomponent food systems, to carry out an integral assessment of the balance of the designed food products. The aim of the study is to study the nutritional composition and optimization of the formulation of minced meat with a filler in the form of a protein-carbohydrate composition of a combined composition by means of design that regulates the stages of creating food products with specified functional properties. At the same time, the chemical, amino acid and fatty acid composition of prototypes of minced meat with fillers of various percentages was investigated in comparison with the control sample. This article presents the results of studies of minced meat with the replacement of meat raw materials with a filler in the amount of 10-25 %. As a result, the optimal recipe for a balanced composition of minced meat with a protein-carbohydrate composition in the amount of 10 % was determined. A mathematical model of a semi-finished meat product with increased nutritional and biological value was developed.

Comparative Analysis of Collagen-Containing Waste Biodegradation, Amino Acid, Peptide and Carbohydrate Composition of Hydrolysis Products

This paper aimed to study the biodegradation of collagen-containing waste (pork skin) induced by collagenase and Neutrase 1.5 MG enzymes and compare the amino acid, peptide, and carbohydrate composition of hydrolysis products. It was found that the degree of biodegradation of collagen-containing raw materials (pork skin) reached 78% when using an enzyme preparation (collagenase with a concentration of 250 U/g of the substrate) at pH 7.0, 40 °C, and a 360 min process duration. It was shown that the content of peptides with a molecular weight of 6.5–14.0 kDa in the hydrolysis products (collagenase) of collagen-containing wastes was 13.4 ± 0.40%, while in the products of hydrolysis (Neutrase 1.5 MG) it was 12.8 ± 0.38%. The study found that the hydrolysis products (Neutrase 1.5 MG) of collagen-containing raw materials contain fewer hexoses, free hexosamines, and hyaluronic acid than the hydrolysis products (collagenase) of collagen-containing raw materials. The content of chondroitin sulfates is practically the same in all samples of hydrolysis products. Proteases with collagenolytic activity are widely used in industry. Recently, they have increasingly been used in pharmaceutical, food, and other industries. Collagenases are promising enzymes for the production of chondroprotectors used for the treatment of osteoarthritis.

Changes in Carbohydrate Composition in Fermented Total Mixed Ration and Its Effects on in vitro Methane Production and Microbiome

The purpose of this experiment was to investigate the changes of carbohydrate composition in fermented total mixed diet and its effects on rumen fermentation, methane production, and rumen microbiome in vitro. The concentrate-to-forage ratio of the total mixed ration (TMR) was 4:6, and TMR was ensiled with lactic acid bacteria and fibrolytic enzymes. The results showed that different TMRs had different carbohydrate compositions and subfractions, fermentation characteristics, and bacterial community diversity. After fermentation, the fermented total mixed ration (FTMR) group had lower contents of neutral detergent fiber, acid detergent fiber, starch, non-fibrous carbohydrates, and carbohydrates. In addition, lactic acid content and relative abundance of Lactobacillus in the FTMR group were higher. Compared with the TMR group, the in vitro ammonia nitrogen and total volatile fatty acid concentrations and the molar proportion of propionate and butyrate were increased in the FTMR group. However, the ruminal pH, molar proportion of acetate, and methane production were significantly decreased in the FTMR group. Notably, we found that the relative abundance of ruminal bacteria was higher in FTMR than in TMR samples, including Prevotella, Coprococcus, and Oscillospira. At the same time, we found that the diversity of methanogens in the FTMR group was lower than that in the TMR group. The relative abundance of Methanobrevibacter significantly decreased, while the relative abundances of Methanoplanus and vadinCA11 increased. The relative abundances of Entodinium and Pichia significantly decreased in the FTMR group compared with the TMR group. These results suggest that FTMR can be used as an environmentally cleaner technology in animal farming due to its ability to improve ruminal fermentation, modulate the rumen microbiome, and reduce methane emissions.

Technological assessment and requirements for quality indicators of Jerusalem artichoke and chicory for processing into inulin and its derivatives

Исследования по разработке требований к показателям качества топинамбура и цикория проведены с использованием отечественных и импортных сортов топинамбура из Костромской обл. (ООО «ВИВА»), цикория из Московской обл. (ООО «Современник») урожая 2017-2018 гг. Исследуемые образцы были проанализированы на показатели качества: массовая доля сухого вещества в соке, стружке; углеводный состав экстракта, полученного горячей водной диффузией стружки топинамбура или цикория; углеводный состав в пересчете в г/на 100 г инулинсодержащего сырья. Установлено, что свежеубранные клубни топинамбура содержат 14-19 % углеводов, в том числе 12-17 % инулина, 1,0-1,6 % ди- и моносахаридов; корнеплоды цикория - 17-19 % углеводов, в том числе 15-17 % инулина, 1,0-2,4 % ди- и моносахаридов. В результате разработаны требования к инулинсодержащему сырью для переработки на инулин: массовая доля сухого вещества в клубнях топинамбура и корнеплодах цикория - не менее 22 %, массовая доля инулина - не менее 14 %, массовая доля ди- и моносахаридов - не более 2 %. Разработана принципиальная универсальная технологическая схема переработки инулинсодержащего сырья на инулин и его производные - олигофруктозу и фруктозный сироп. Определены технологические режимы проведения непрерывной диффузии из стружки сырья: температура 80…85 °С, продолжительность процесса - 1 ч, величина гидромодуля - 1:2. Разработаны методы очистки инулинсодержащего экстракта: кислотная коагуляция, очистка активным углем, двухступенчатая ионообменная очистка по схеме К1+А1+К2+А2. В результате очистки примеси сиропа снижаются в десятки раз: цветность - не более 0,5 ед. опт. пл.; массовая доля протеина - не более 0,5 %; золы - не более 0,2 %. Для получения олигофруктозы или фруктозного сиропа предложено проводить неполный или полный гидролиз очищенного сиропа с использованием ферментных препаратов инулиназы компании «Новозаймс» (Дания). Определены оптимальные условия гидролиза инулина для получения олигофруктозы: Т=55…58 °С; рН 4,7-5,2; СВ=19-20 % при продолжительности 22-24 ч и дозировке препарата 0,3-0,4 ед. INU/г СВ. Разработанная технология позволит создать отечественные продукты рационального оздоровительно-профилактического, диетического и диабетического назначения для функционального питания населения. Studies on the development of requirements for the quality indicators of Jerusalem artichoke and chicory were carried out using domestic and imported varieties of Jerusalem artichoke from the Kostroma region. (LLC «VIVA»), chicory from the Moscow region. (LLC «Sovremennik») harvest 2017-2018 The samples under study were analyzed for their quality indicators: mass fraction of dry matter in juice, shavings; carbohydrate composition of the extract obtained by hot water diffusion of Jerusalem artichoke and chicory; carbohydrate composition in terms of g / per 100 g of inulin-containing raw materials. It was found that freshly harvested Jerusalem artichoke tubers contain 14-19 % carbohydrates, including 12-17 % inulin, di- and monosaccharides 1,0-1,6 %; chicory roots 17- 19 % carbohydrates, including 15-17 % inulin, 1,0-2,4 % di- and monosaccharides. As a result, requirements were developed for inulin-containing raw materials for processing into inulin: mass fraction of dry matter in Jerusalem artichoke tubers and chicory root crops - at least 25 %; mass fraction of inulin - not less than 14%; mass fraction of di- and monosaccharides - no more than 2 %. A basic universal technological scheme for processing inulin-containing raw materials into inulin and its derivatives - oligofructose and fructose syrup has been developed. The technological modes of continuous diffusion from raw material chips were determined: temperature 80…85 °С, process duration - 1 h, hydromodule value - 1:2. Methods for purification of inulin-containing extract have been developed - acid coagulation, purification with active carbon, two-stage ion-exchange purification according to the scheme K1 + A1 + K2 + A2. As a result of purification, syrup impurities are reduced tenfold: color - no more than 0,5 units. wholesale pl.; mass fraction of protein - no more than 0,5 %; ash - no more than 0,2 %. To obtain oligofructose or fructose syrup, it is proposed to carry out incomplete or complete hydrolysis of purified syrup using inulinase enzyme preparations from Novozymes (Denmark). The optimal conditions for the hydrolysis of inulin to obtain oligofructose have been determined: T=55…58 °С; pH 4,7-5,2; DM=19-20% with a duration of 22-24 hours and a dosage of the drug 0,3-0,4 units. INU/g DM. The developed technology will make it possible to create domestic products of rational health-improving, preventive, dietary and diabetic nutrition for the creation of functional nutrition products for the population.

Development of Lactic acid Microorganisms during Fermentation of Substrate with an Increased Concentration of Carbohydrates

Introduction. Creating favorable conditions for the development of lactic acid microorganisms is one of the main factors in obtaining high-quality fermented products. The cycle of their life directly depends on the amount and composition of carbohydrates in plant tissue. Since a significant part of carbohydrates is consumed at the initial stage of fermentation process, additional fortification is needed. The research objective was to study the development rate of lactic acid microorganisms during the fermentation of plant substrate with a modified carbohydrate composition. Study objects and methods. The research featured model medium based on white cabbage of the Parus variety. The medium was fermented with different strains of lactic acid microorganisms: at the first stage of fermentation – Leuconostoc mesenteroides, at the second stage – Lactobacillus casei VKM 536, Lactobacillus plantarum VKM B-578, Lactobacillus brevis VKM B-1309, and their paired consortia. The initial plant material was subjected to grinding and removal of native microflora for the development of target lactic acid microorganisms, then inoculated with L. mesenteroides. The target lactic acid microorganisms were introduced after the first stage of fermentation with simultaneous adjustment of the carbohydrate composition. Results and discussion. The technology included modes of controlled two-stage microbial transformation of plant raw materials using modification of the carbohydrate composition of the substrate. A number of experiments made it possible to select the optimal composition of the consortium and establish the optimal fermentation time at the main stage of microbial processing. When the plant substrate was fermented by the consortium of L. casei + L. plantarum with an increased carbohydrate component, the decrease in the concentration was quite small: after 5–30 days, the decrease in the concentration of microorganisms did not exceed one order of magnitude, which was insignificant at an initial concentration of eight orders of magnitude. In other consortia, the decrease in the concentration of microorganisms was more pronounced. Conclusion. The fortification of the vegetable substrate with carbohydrates made it possible to maintain the concentration of lactic acid microorganisms at a level comparable to the concentration at the time of inoculation. The concentrations of microorganisms varied slightly in both monocultures and their paired consortia during the entire main fermentation stage of the model medium with a modified carbohydrate component. By the end of the main fermentation stage, the concentration of microorganisms did not fall below 107 CFU/g. Therefore, the resulting system “microflora – substrate” proved to have probiotic properties. The study can be used to develop new technological modes of controlled step-by-step fermentation of plant raw materials in order to improve the quality indicators of the final product.

INFLUENCE OF HYDROLYTIC TREATMENT ON THE CONTENT OF REDUCING SUBSTANCES IN NEUTRAL-SULPHITE ALKALI

The aim of this work is to develop a technology for the preparation of neutral-sulfite liquors formed during the production of fibrous semi-finished products - cellulose from birch wood - for subsequent use as a nutrient medium for the cultivation of microorganisms. Acid hydrolysis was carried out at a temperature of 100 °С at a ratio of a 10% sulfuric acid solution to a liquor sample of 1 : 1. Enzymatic hydrolysis of neutral sulfite liquors was carried out with the enzyme preparations Accellerase XY and Accellerase XC at 50±2 °C and 60±2 °C. The end of hydrolysis was determined by the cessation of the increase in the content of reducing substances (RS) in the hydrolyzate. The original neutral sulphite lye contained 9.4% dry matter, 21.7 g/l of reducing substances, pH 5.3±0.2. It has been shown that as a result of enzymatic hydrolysis, the content of insoluble dry residue in the hydrolyzate decreases to 8.32% and 8.41%, respectively, and during acid hydrolysis – to 7.8%. The content of RS in neutral sulfite lye after acid hydrolysis increases by an average of 3 times, while after enzymatic hydrolysis - a maximum of 2 times. It was found by gas-liquid chromatography that pentoses predominate in the obtained hydrolysates. Microbiological processing of media with a similar carbohydrate composition is possible by a number of strains of microorganisms capable of assimilating pentoses, for example, yeast-like fungi of the Saccharomycetaceae family and bacteria of the Enterobacteriaceae family.