Objective: The aims of this study were to reveal the magnitude of the differences in protein structures at a cellular level as well as protein utilization and availability among soybean meal (SBM), canola meal (CM), and rapeseed meal (RSM) as feedstocks in China. Methods: Experiments were designed to compare the three different types of feedstocks in terms of: i) protein chemical profiles; ii) protein fractions partitioned according to Cornell Net Carbohydrate and Protein System; iii) protein molecular structures and protein second structures; iv) special protein compounds-amino acid (AA); v) total digestible protein and energy values; vi) in situ rumen protein degradability and intestinal digestibility. The protein second structures were measured using FT/IR molecular spectroscopy technique. A summary chemical approach in National Research Council (NRC) model was applied to analyze truly digestible protein. Results: The results showed significant differences in both protein nutritional profiles and protein structure parameters in terms of ${\alpha}-helix$, ${\beta}-sheet$ spectral intensity and their ratio, and amide I, amide II spectral intensity and their ratio among SBM, CM, and RSM. SBM had higher crude protein (CP) and AA content than CM and RSM. For dry matter (DM), SBM, and CM had a higher DM content compared with RSM (p<0.05), whereas no statistical significance was found between SBM and CM (p = 0.28). Effective degradability of CP and DM did not demonstrate significant differences among the three groups (p>0.05). Intestinal digestibility of rumen undegradable protein measured by three-step in vitro method showed that there was significant difference (p = 0.05) among SBM, CM, and RSM, which SBM was the highest and RSM was the lowest with CM in between. NRC modeling results showed that digestible CP content in SBM was significantly higher than that of CM and RSM (p<0.05). Conclusion: This study suggested that SBM and CM contained similar protein value and availability for dairy cattle, while RSM had the lowest protein quality and utilization.
A simple, fast and reproducible quantitative analysis method for sugar concentration composed in oligosaccharide mixture was developed. Two glass TLC plates were prepared per sample. After dipping one plate into the copper bicinchoninate reagent and the other plate into 5% sulfuric acid solution, both plates were baked in microwave oven until sugar spots were developed or the surface temperature of TLC plate becomes 60 to 70 $^{\circ}C$. The corrective factor values [F value =(the value of total sugar concentration converted as glucose unit/the value of reducing sugar concentration converted as glucose unit)/(polymerization degree of sugar)] of different molecular weight sugars were determined. Within the concentration of 0.25∼1.0 $\mu\textrm{g}$ in each sample loaded, the fructose-F (corrective factor value of fructose) was 0.45, yet for the higher concentration (2.5∼7.5 $\mu\textrm{g}$) fructose-F was 1.0. In case of glucose, in the range of 0.5∼7.5 $\mu\textrm{g}$, glucose-F was same as fructose-F, 1.0. However, as the molecular weight of sugar was increased, the F values were decreased in both maltodextrin and isomaltodextrin oligosaccharides in 0.5∼7.5 $\mu\textrm{g}$ of each sample loaded. Interestingly, F values were equal for the same molecular weight sugars, although the structures were different from each other. Using F value of each sugar, we could determine and compare the exact total sugar concentration of different molecular weight maltooligosaccharide and isomaltooligosaccharide. We also could determine if the unknown sugar was a reducing or non-reducing compound by using optimized TLC with microwave oven method.
The innate immune system is important for the first line of host defence against infectious agents, which have penetrated the mechanical barriers. Mannose-binding lectin (MBL or mannan-binding protein, MBP) is a serum protein that is synthesized in the liver as a part of the acute phase response. MBL binds to carbohydrate structures presented by a wide range of pathogenic bacteria, viruses, fungi, and parasites. MBL is synthesized as a monomer that has a carboxy-terminal carbohydrate recognition domain, a neck region and a collagen region. Low MBL level was reported to be the most frequent immuno-deficiency syndrome. Although extensive studies have yielded detailed information on the structure of MBL, functions of the MBL complex are not fully understood yet. We, here, present cloning process of MBL cDNA from the rat liver and production of truncated recombinant MBL protein using a bacterial expression system in order to produce anti-MBL polyclonal antibody. Anti-MBL polyclonal antibody was raised in a New Zealand rabbit and its affinity was tested against recombinant protein using western blot technique. MBL cDNA, recombinant protein and anti-MBL antibody could be used as great arsenals to dissect cellular biochemistry of MBL.
Journal of the Korean Applied Science and Technology
/
v.32
no.3
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pp.546-567
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2015
Silicone-based surfactants consist of a hydrophobic organosilicone group coupled to one or more hydrophilic polar groups, while the hydrophobic groups of hydrocarbon surfactants are hydrocarbons. Silicone surfactants have been widely used in many industrial fields starting from polyurethane foam to construction materials, cosmetics, paints & inks, agrochemicals, etc., because of their low surface tension, lubricity, spreading, water repellency and thermal and chemical stability. A wide range of silicone surfactant structures are required to provide the functional diversity for reflecting the necessities in the various applications. This review covers the basic properties and the synthetic schemes of polydimethylsiloxane and reactive polysiloxanes as hydrophobic siloxane backbones, the main reaction schemes, such as hydrosilylation reaction, for coupling reactive polysiloxanes to hydrophilic groups, and the synthetic schemes of the main polysiloxane surfactants including polyether-, ionic-, carbohydrate-type surfactants.
Glycoproteins PAS-6(50 kDa) and -7(47 kDa) from the bovine milk fat globule membrane share a common protein core but differ in their carbohydrate moiety. We have analyzed and proposed the structures of the N-linked sugar chains of PAS-7 by Oxford Glyco System(OGS) RAAM2000. The N-linked sugar chains were liberated from PAS-7 by hydrazinolysis and, after modifying the reducing ends with 2-aminobenzamide(2-AB), were separated into one neutral(7N, 55%) and two acidic(7M, mono-, 43%; 7D, di-, 2%) sugar chain groups. 7N was finally separated into 5 chains(a, b, c, d, and e), respectively. The structure of this 2AB-neutral sugar chain was determined by sugar analysis, exoglycosidase digestion with OGS glycosidase Kit and OGS RAAM2000 system. The results show that fraction e was the same of reported 7N1A, the biantennary complex type with a fucose on reducing end and two N-acetyllactosamine branch on non-reducing end. Therefore, it was proved that OGS RAAM2000 method is in conformity with conventional analysis of sugar chain structure from bovine PAS-7.
Soybean contains a high concentration of carbohydrates that consist mainly of non-starch polysaccharides (NSP) and oligosaccharides. The NSP can be divided into insoluble NSP (mainly cellulose) and soluble NSP (composed mainly of pectic polymers, which are partially soluble in water). Monogastric animals do not have the enzymes to hydrolyze these carbohydrates, and thus their digestion occurs by means of bacterial fermentation. The fermentation of soybean carbohydrates produces short chain fatty acids that can be used as an energy source by animals. The utilization efficiency of the carbohydrates is related to the chemical structure, the level of inclusion in the diet, species and age of the animal. In poultry, soluble NSP can increase digesta viscosity, reduce the digestibility of nutrients and depress growth performance. In growing pigs, these effects, in particular the effect on gut viscosity, are often not so obvious. However, in weaning piglets, it is reported that soy oligosaccharides and soluble NSP can cause detrimental effects on intestinal health. In monogastrics, consideration must be given to the anti-nutritive effect of the NSP on nutrient digestion and absorption on one hand, as well as the potential benefits or detriments of intestinal fermentation products to the host. This mirrors the needs for i) increasing efficiency of utilization of fibrous materials in monogastrics, and ii) the maintenance and improvement of animal health in antibiotic-free production systems, on the other hand. For example, ethanol/water extraction removes the low molecular weight carbohydrate fractions, such as the oligosaccharides and part of the soluble pectins, leaving behind the insoluble fraction of the NSP, which is devoid of anti-nutritive activities. The resultant product is a high quality soy protein concentrate. This paper presents the composition and chemical structures of carbohydrates present in soybeans and discusses their nutritive and anti-nutritive effects on digestion and absorption of nutrients in pigs and poultry.
Fluorescence properties and carbohydrate content were investigated using ultrafiltrated size fractions of dissolved organic matters (DOM) originated from different sources. The materials included a treated sewage, an algal organic matter, and a soil leachate, all of which are major constituents of dissolved organic matter in a typical urban river. Four different size fractions were separated from the three sources of each DOM. The size distribution demonstrated that a higher molecular weight fraction was more present in soil leachate compared to two other source DOMs. A higher content of carbohydrates was observed in the following order - algal DOM > treated sewage > soil leachate. A wide range of specific UV absorbance was observed from size fractions of a single source DOM, indicating that aromatic carbon structures are heterogeneously distributed within one source of DOM. The structural heterogeneity was the most pronounced for the soil leachate. The fluorescence index ($F_{450}/F_{500}$) of the treated sewage was similar to that (2.0) typically obtained from autochthonous DOM, suggesting that the treated sewage exhibited autochthonous organic matter-like properties. No protein-like fluorescence intensities were observed for all of the soil leachate size fractions whereas they were observed with two other source DOMs. Based upon the fluorescence peak ratios from fluorescence excitation-emission matrix (EEM), two discrimination indices could be suggested to distinguish three different source DOMs. It is expected that the suggested discrimination indices will be useful to predict the sources of DOM in a typical urban river affected by treated sewage.
Exopolysaccharides (EPSs) such as capsular polysaccharide (CPS) are important bioactive carbohydrate compounds and are often used as bioenrichment agents and bioabsorbers to remove environmental pollutants like di-n-butyl phthalate (DBP). Among the EPS-producing bacteria, lactic acid bacteria (LAB) have gained the most attention. As generally recognized as safe (GRAS) microorganisms, LAB can produce EPSs having many different structures and no health risks. However, EPS production by LAB does not meet the needs of large-scale application on an industrial scale. Here, the capA gene (encoding CPS biosynthesis protein) was overexpressed in Lactobacillus plantarum P1 to improve the production of EPSs and further enhance the DBP adsorption capability. Compared with P1, the CPS production in capA overexpressed strain was increased by 11.3 mg/l, and the EPS thickness was increased from 0.0786 ± 0.0224 ㎛ in P1 to 0.1160 ± 0.0480 ㎛ in P1-capA. These increases caused the DBP adsorption ratio of P1-capA to be doubled. Overall, the findings in this study provide a safe method for the adsorption and removal of DBP.
Joon-hui Chung;Jehyeong Yeon;Hoon Je Seong;Si-Hyun An;Da-Yeon Kim;Younggun Yoon;Hang-Yeon Weon;Jeong Jun Kim;Jae-Hyung Ahn
Journal of Microbiology and Biotechnology
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v.32
no.12
/
pp.1561-1572
/
2022
Plastic pollution has been recognized as a serious environmental problem, and microbial degradation of plastics is a potential, environmentally friendly solution to this. Here, we analyzed and compared microbial communities on waste plastic films (WPFs) buried for long periods at four landfill sites with those in nearby soils to identify microbes with the potential to degrade plastics. Fourier-transform infrared spectroscopy spectra of these WPFs showed that most were polyethylene and had signs of oxidation, such as carbon-carbon double bonds, carbon-oxygen single bonds, or hydrogen-oxygen single bonds, but the presence of carbonyl groups was rare. The species richness and diversity of the bacterial and fungal communities on the films were generally lower than those in nearby soils. Principal coordinate analysis of the bacterial and fungal communities showed that their overall structures were determined by their geographical locations; however, the microbial communities on the films were generally different from those in the soils. For the pulled data from the four landfill sites, the relative abundances of Bradyrhizobiaceae, Pseudarthrobacter, Myxococcales, Sphingomonas, and Spartobacteria were higher on films than in soils at the bacterial genus level. At the species level, operational taxonomic units classified as Bradyrhizobiaceae and Pseudarthrobacter in bacteria and Mortierella in fungi were enriched on the films. PICRUSt analysis showed that the predicted functions related to amino acid and carbohydrate metabolism and xenobiotic degradation were more abundant on films than in soils. These results suggest that specific microbial groups were enriched on the WPFs and may be involved in plastic degradation.
For the purpose of making clear the activity of the alkaline phosphatase to the morphogenesis and function of the various structures of the developing chick kidney in relation to PAS-positive materials and phospholipid, the author observed histochemically the aforementioned enzyme and other substances. The mesonephros and metanephros of 4-20 day white leghorn embryos were used, obtaining the following results. 1. Before definite appearance of the secretory tubules the alkaline phosphatase activity showed strongly in the undifferentiated mesenchymal tissue. As the tubules grew differentiated, the alkaline phosphatase activity was found to have disappeared in the mesenchymal tissue surrounding the tubules. The above mentioned fact indicates that the alkaline phosphatase may be concerned with morphogenesis of the developing chick kindney. 2. The fact that the strong alkaline phosphatase activity and the occurrence of the PAS-positive materials were observed at the luminal borders of differentiated secretory tubules of mesonephros and metanephros, indicates that alkaline phosphatase may be concerned with reabsorption of carbohydrate at the borders. 3. A strong positive reaction of phospholipid was found in the cytoplasm and brush borders of the mesonephric and metanephric tubules. The fact that vicissitude of alkaline phosphatase was found to coincide with that of phospholipid suggests that the enzyme may have influence on the metabolism of the phospholipid.
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