Gelatin based preservation technologies on the quality of food: a comprehensive review.

Gelatin has played a great potential in food preservation because of its low price and superior film forming characteristics. This review provides a comprehensive overview of the latest research progress and application of gelatin preservation technologies (film, coating, antifreeze peptide, etc.), discussing their preservation mechanisms and efficiency through the viewpoints of quality and shelf life of animal and aquatic products as well as fruits and vegetables. It showed that bioactive and intelligent gelatin-based films exhibit antibacterial, antioxidant, water resistance and pH responsive properties, making them excellent for food preservation. In addition, pH responsive properties of films also intuitively reflect the freshness of food by color. Similarly, gelatin and its hydrolysate can be widely used in antifreeze peptides to reduce the mass loss of food during freezing and extend the shelf life of frozen food. However, extensive works are still required to extend their commercial application values.

Recent progress in the structure of glycogen serving as a durable energy reserve in bacteria.

Glycogen is conventionally considered as a transient energy reserve that can be rapidly synthesized for glucose accumulation and mobilized for ATP production. However, this conception is not completely applicable to prokaryotes due to glycogen structural heterogeneity. A number of studies noticed that glycogen with small average chain length gc in bacteria has the potential to degrade slowly, which might prolong bacterial environment survival. This phenomenon was previously examined and later formulated as the durable energy storage mechanism hypothesis. Although recent research has been warming to the hypothesis, experimental validation is still missing at current stage. In this review, we summarized recent progress of the hypothesis, provided a supporting mathematical model, and explored the technical pitfalls that shall be avoided in glycogen study.

Molecular Structure of Glycogen in Escherichia coli.

Glycogen, a randomly branched glucose polymer, provides energy storage in organisms. It forms small ß particles which in animals bind to form composite α particles, which give better glucose release. Simulations imply ß particle size is controlled only by activities and sizes of glycogen biosynthetic enzymes and sizes of polymer chains. Thus, storing more glucose requires forming more ß particles, which are expected to sometimes form α particles. No α particles have been reported in bacteria, but the extraction techniques might have caused degradation. Using milder glycogen extraction techniques on Escherichia coli, transmission electron microscopy and size-exclusion chromatography showed α particles, consistent with this hypothesis for α-particle formation. Molecular density and size distributions show similarities with animal glycogen, despite very different metabolic processes. These general polymer constraints are such that any organism which needs to store and then release glucose will have similar α and ß particle structures: a type of convergent evolution.

Molecular structure of glycogen in diabetic liver.

Liver glycogen (involved in maintaining blood-sugar levels) is a hyperbranched glucose polymer containing ß particles (diameter ~20 nm), which can form composite α particles (diameter ~50-300 nm), and includes a small but significant amount of bound protein. Size distributions of glycogen from livers of healthy and diabetic mice were examined using size-exclusion chromatography with two separate eluents: aqueous eluent and dimethylsulfoxide (DMSO) eluent. Morphologies were examined with transmission electron microscopy. Diabetic glycogen (DG) exhibited many α particles in the mild water-based solvent, but in DMSO, which breaks H bonds, these degraded to ß particles; α particles however were always present in healthy glycogen (HG). This DG fragility shows the binding of ß into α particles is different in HG and DG. The diabetic α particle fragility may be involved with the uncontrolled blood-sugar release symptomatic of diabetes: small ß particles degrade more easily to glucose than α particles. This has implications for diabetes management.

Dietary xylo-oligosaccharides and arabinoxylans improved growth efficiency by reducing gut epithelial cell turnover in broiler chickens.

BACKGROUND: One of the main roles of the intestinal mucosa is to protect against environmental hazards. Supplementation of xylo-oligosaccharides (XOS) is known to selectively stimulate the growth of beneficial intestinal bacteria and improve gut health and function in chickens. XOS may have an impact on the integrity of the intestinal epithelia where cell turnover is critical to maintain the compatibility between the digestive and barrier functions. The aim of the study was to evaluate the effect of XOS and an arabinoxylan-rich fraction (AXRF) supplementation on gut function and epithelial integrity in broiler chickens. METHODS: A total of 128 broiler chickens (Ross 308) were assigned into one of two different dietary treatments for a period of 42 d: 1) control diet consisting of a corn/soybean meal-based diet; or 2) a control diet supplemented with 0.5% XOS and 1% AXRF. Each treatment was randomly distributed across 8 pens (n = 8) with 8 chickens each. Feed intake and body weight were recorded weekly. On d 42, one male chicken per pen was selected based on average weight and euthanized, jejunum samples were collected for proteomics analysis. RESULTS: Dietary XOS/AXRF supplementation improved feed efficiency (P < 0.05) from d 1 to 42 compared to the control group. Proteomic analysis was used to understand the mechanism of improved efficiency uncovering 346 differentially abundant proteins (DAP) (Padj < 0.00001) in supplemented chickens compared to the non-supplemented group. In the jejunum, the DAP translated into decreased ATP production indicating lower energy expenditure by the tissue (e.g., inhibition of glycolysis and tricarboxylic acid cycle pathways). In addition, DAP were associated with decreased epithelial cell differentiation, and migration by reducing the actin polymerization pathway. Putting the two main pathways together, XOS/AXRF supplementation may decrease around 19% the energy required for the maintenance of the gastrointestinal tract. CONCLUSIONS: Dietary XOS/AXRF supplementation improved growth efficiency by reducing epithelial cell migration and differentiation (hence, turnover), actin polymerization, and consequently energy requirement for maintenance of the jejunum of broiler chickens.

Liver glycogen fragility in the presence of hydrogen-bond breakers.

Glycogen, a complex branched glucose polymer, is responsible for sugar storage in blood glucose homeostasis. It comprises small ß particles bound together into composite α particles. In diabetic livers, α particles are fragile, breaking apart into smaller particles in dimethyl sulfoxide, DMSO; they are however stable in glycogen from healthy animals. We postulate that the bond between ß particles in α particles involves hydrogen bonding. Liver-glycogen fragility in normal and db/db mice (an animal model for diabetes) is compared using various hydrogen-bond breakers (DMSO, guanidine and urea) at different temperatures. The results showed different degrees of α-particle disruption. Disrupted glycogen showed changes in the mid-infra-red spectrum that are related to hydrogen bonds. While glycogen α-particles are only fragile under harsh, non-physiological conditions, these results nevertheless imply that the bonding between ß particles in α particles is different in diabetic livers compared to healthy, and is probably associated with hydrogen bonding.

Oxidative stress induced by plasma-activated water stimulates astaxanthin production in Phaffia rhodozyma.

Astaxanthin is used extensively in the nutraceutical, aquaculture, and cosmetic industries. The current market necessitates higher astaxanthin production from Phaffia rhodozyma (P. rhodozyma) due to its higher cost compared to chemical synthesis. In this study, a bubble discharge reactor was developed to generate plasma-activated water (PAW) to produce PAW-made yeast malt (YM) medium. Due to oxidative stress induced by PAW, strains cultured in 15 and 30 min-treated PAW-made medium produced 7.9 ± 1.2 % and 12.6 ± 1.4 % more carotenoids with 15.5 ± 3.3 % and 22.1 ± 1.3 % more astaxanthin, respectively. Reactive oxygen species (ROS) assay results showed that ROS generated by plasma-water interactions elevated intracellular ROS levels. Proteomic analysis revealed increased expression of proteins involved in the cellular response to oxidative stress as well as carotenoid biosynthesis, both of which contribute to higher yields of astaxanthin. Overall, this study supports the potential of PAW to increase astaxanthin yields for industrial-scale production.

A Non-Degradative Extraction Method for Molecular Structure Characterization of Bacterial Glycogen Particles.

Currently, there exist a variety of glycogen extraction methods, which either damage glycogen spatial structure or only partially extract glycogen, leading to the biased characterization of glycogen fine molecular structure. To understand the dynamic changes of glycogen structures and the versatile functions of glycogen particles in bacteria, it is essential to isolate glycogen with minimal degradation. In this study, a mild glycogen isolation method is demonstrated by using cold-water (CW) precipitation via sugar density gradient ultra-centrifugation (SDGU-CW). The traditional trichloroacetic acid (TCA) method and potassium hydroxide (KOH) method were also performed for comparison. A commonly used lab strain, Escherichia coli BL21(DE3), was used as a model organism in this study for demonstration purposes. After extracting glycogen particles using different methods, their structures were analyzed and compared through size exclusion chromatography (SEC) for particle size distribution and fluorophore-assisted capillary electrophoresis (FACE) for linear chain length distributions. The analysis confirmed that glycogen extracted via SDGU-CW had minimal degradation.

Starch molecular fine structure is associated with protein composition in chickpea seed.

Chickpea (Cicer arietinum L.) seed is a nutritional food high in starch and protein. This study aims to find the relationships between the molecular fine structure of starch and the composition of storage proteins and metabolic enzymes, using different chickpea varieties. It is found that storage proteins and starch biosynthetic enzymes influence each other. The initial formation of amylopectin molecules is affected by storage proteins, as suggested by the positive correlation (p < 0.01) between the average molecular size of amylopectin and total protein content. In addition, a higher amount of seed globulin could be an indication of higher amylose content and more short - medium amylose chains (degree of polymerization, DP, 118-2000). This study might assist selection of chickpea varieties with desirable qualities, such as low starch digestibility.

Effects of fasting on liver glycogen structure in rats with type 2 diabetes.

Liver glycogen, a highly branched glucose polymer, is important for blood sugar homeostasis. It comprises α particles which are made of linked ß particles; the molecular structure changes diurnally. In diabetic liver, the α particles are fragile, easily breaking apart into ß particles in chaotropic agents such as dimethyl sulfoxide. We here use size-exclusion chromatography to study how fasting changes liver-glycogen structure in vivo for mice in which type-2 diabetes had previously been induced. Diabetic glycogen degraded enzymatically more quickly in the fasted animals than did glycogen without fasting, with fewer α particles, which however were still fragile. The glycogen had fewer long chains and more shorter chains after fasting. This study gives an overview of the in vivo dynamic changes in α-particles under starvation conditions in both normal and diabetic livers.

Some molecular structural features of glycogen in the kidneys of diabetic rats.

Glycogen, a highly-branched glucose polymer, functions as a sugar reservoir in many organs and tissues. Liver glycogen comprises small ß particles which can bind to form into large agglomerates (α particles) which readily degrade to ß particles in diabetic livers. Muscle glycogen has only ß particles, optimal for quick energy release. Healthy kidney contains negligible glycogen, but there is an abnormally high accumulation in diabetic kidneys. We here compare the molecular structure of glycogen in diabetic kidneys with that in liver and muscle, using a diabetic rat model. This involved exploring extraction techniques to minimize glycogen degradation. Using size exclusion chromatography and transmission electron microscopy, it was found that there were only ß particles in diabetic kidneys. These are postulated to form during periods of abnormally high blood sugar, the driving force being the need to reduce blood sugar under such circumstances.

Glycogen structure in type 1 diabetic mice: Towards understanding the origin of diabetic glycogen molecular fragility.

Glycogen is a complex branched glucose polymer. Liver glycogen in db/db mouse, a type-2 diabetic mouse model, has been found to be more molecularly fragile than in healthy mice. Size-exclusion chromatography was employed in this study to investigate the molecular structure of liver glycogen in two types of type 1 diabetic mouse models (NOD and C57BL/6J mice), sacrificed at various times throughout the diurnal cycle, and the fragility of liver glycogen after exposure to a hydrogen-bond disruptor were tested. Type 1 diabetic mice exhibit a similar glycogen fragility with that observed for db/db mice. This eliminates many of the potential causes for glycogen molecular fragility; the most likely explanation is that it is caused by high blood-glucose level and/or insulin deficiency, both phenotypes being common to both type 1 and type 2 diabetic mice. This result suggests ways towards new drug targets for the management of diabetes.

Proteomic Investigation of the Binding Agent between Liver Glycogen ß Particles.

Glycogen is a highly branched glucose polymer which plays an important role in glucose storage and the maintenance of blood sugar homeostasis. The dimeric protein glycogenin can self-glucosylate to act as a primer for glycogen synthesis, eventually resulting in small (∼20 nm diameter) glycogen ß particles with a dimer of glycogenin at their core. In the liver, glycogen is also found in the form of α particles: large bound composites of many ß particles. Here, we provide evidence using qualitative and quantitative proteomics and size-exclusion chromatography from healthy rat, mouse, and human liver glycogen that glycogenin is the binding agent linking ß particles together into α particles.

Exploring glycogen biosynthesis through Monte Carlo simulation.

Glycogen, a complex branched polymer of glucose (average chain length ~10 monomer units), is the blood-sugar reservoir in humans and other animals. Certain aspects of its molecular structure relevant to its biological functions are currently unamenable to experimental exploration. Knowledge of these is needed to develop future models for quantitative data-fitting to obtain mechanistic understanding of the biosynthetic processes that give rise to glycogen structure. Monte Carlo simulations of the biosynthesis of this structure with realistic macromolecular parameters reveal how chain growth and stoppage (the latter assumed to be through both the action of glycogen branching enzyme and other degradative enzymes, and by hindrance) control structural features. The simulated chain-length, pair-distance and radial density distributions agree semi-quantitatively with the limited available data. The simulations indicate that a steady state in molecular structure and size is rapidly obtained, that molecular density reaches a maximum near the center of the particle (not at the periphery, as is the case with dendrimers), and that particle size is controlled by both enzyme activity and hindrance. This knowledge will aid in the understanding of diabetes (loss of blood-sugar control), which has been found to involve subtle differences in glycogen molecular structure.

Diurnal changes of glycogen molecular structure in healthy and diabetic mice.

Glycogen is a complex branched glucose polymer functioning as a blood-sugar reservoir in animals. Liver glycogen ß particles can bind together to form α particles, which have a slower enzymatic degradation to glucose. The linkage between ß particles in α particles in diabetic liver breaks (is fragile) in dimethyl sulfoxide (DMSO), a H-bond disruptor, consistent with blood-sugar homeostasis loss in diabetes. We examined diurnal changes in the molecular structure of healthy and diabetic mouse-liver glycogen. Healthy mouse glycogen was fragile to DMSO during glycogen synthesis but not degradation; diabetic glycogen was always fragile. Two alternative mechanisms for this are suggested: healthy glycogen is fragile when formed and becomes stable during subsequent degradation, a process damaged in diabetes; alternatively, there are two types of glycogen: one compact but fragile and the other loose but non-fragile. This suggests potential types of diabetes drug targets through modifying the activities of glycogen synthesis enzymes.

Genetic and Proteomic characterization of Bile Salt Export Pump (BSEP) in Snake Liver.

Snake gallbladder, a traditional Chinese medicine, has been believed in various Asian countries to improve visual acuity and alleviate rheumatism. Bile acids, a major component of the gallbladder, are toxic to the liver and kidney in humans and animals due to its detergent effects, while also exhibiting therapeutic effects due to an increase in the gallbladder contractions of muscle strips in patients with cholesterol gallstones. Secretion of bile acids in human and mammals depends on the bile salt export pump (BSEP), a liver-specific adenosine triphosphate (ATP)-binding cassette transporter encoded by ABCB11. However, the presence of BSEP in snakes has not been thoroughly explored. Here we confirm the existence of BSEP and its coding DNA sequence in snakes on both the proteomic and genetic level. This work provides information on the snake ABCB11 sequence and helps further potential genetic manipulation to affect bile salt metabolism. Our study provides the foundation for research on bile acid production from snakes by using modern genetic and proteomic methodologies.

Relationships between protein content, starch molecular structure and grain size in barley.

Correlations among barley protein, starch molecular structure and grain size were determined using 30 barley samples with variable protein contents. Starch molecular structure was characterized by fluorophore-assisted carbohydrate electrophoresis and by size-exclusion chromatography (SEC, also termed GPC). The chain-length distributions of amylopectin were fitted using a mathematical model reflecting the relative activities of starch branching enzymes and starch synthase enzymes. Increased protein content significantly and negatively correlated with higher amounts of amylose with longer chains (degree of polymerization, DP 1600-40000) while barley grain sizes positively associated with starch contents. Protein content also positively correlated with the proportion of longer chains of amylopectin (DP 34-100). These results showed that the enzyme activities of starch synthases change with protein content, leading to altered starch contents, structures and grain sizes. From this perspective, selecting for large grain size (or low protein content) does not necessarily relate to starch structure, although may suggest long chains of amylopectin. Measuring starch structure could give a good indication of process performance in human food, animal feed and brewing, as all these structural features contribute to significant functional properties.

A new non-degradative method to purify glycogen.

Liver glycogen, a complex branched glucose polymer containing a small amount of protein, is important for maintaining glucose homeostasis (blood-sugar control) in humans. It has recently been found that glycogen molecular structure is impaired in diabetes. Isolating the carbohydrate polymer and any intrinsically-attached protein(s) is an essential prerequisite for studying this structural impairment. This requires an effective, non-degradative and efficient purification method to exclude the many other proteins present in liver. Proteins and glycogen have different ranges of molecular sizes. Despite the plethora of proteins that might still be present in significant abundance after other isolation techniques, SEC (size exclusion chromatography, also known as GPC), which separates by molecular size, should separate those extraneous to glycogen from glycogen with any intrinsically associated protein(s). A novel purification method is developed for this, based on preparative SEC following sucrose gradient centrifugation. Proteomics is used to show that the new method compares favourably with current methods in the literature.