Coarse cereals modulating chronic low-grade inflammation: review.

Chronic low-grade inflammation (CLGI) is closely associated with various chronic diseases. Researchers have paid attention to the comprehensive application and development of food materials with potential anti-inflammatory activity. Owing to their abundant nutrients and biological activities, coarse cereals have emerged as an important component of human diet. Increasing evidence has revealed their potential protective effects against CLGI in chronic conditions. However, this property has not been systematically discussed and summarized. In the present work, numerous published reports were reviewed to systematically analyze and summarize the protective effects of coarse cereals and their main active ingredients against CLGI. Their current utilization state was investigated. The future prospects, such as the synergistic effects among the active compounds in coarse cereals and the biomarker signatures of CLGI, were also discussed. Coarse cereals show promise as food diet resources for preventing CLGI in diseased individuals. Their active ingredients, including ß-glucan, resistant starch, arabinoxylan, phenolic acids, flavonoids, phytosterols and lignans, function against CLGI through multiple possible intracellular signaling pathways and immunomodulatory effects. Therefore, coarse cereals play a crucial role in the food industry due to their health effects on chronic diseases and are worthy of further development for possible application in modulating chronic inflammation.

Thermostability Improvement of L-Asparaginase from Acinetobacter soli via Consensus-Designed Cysteine Residue Substitution.

To extend the application range of L-asparaginase in food pre-processing, the thermostability improvement of the enzyme is essential. Herein, two non-conserved cysteine residues with easily oxidized free sulfhydryl groups, Cys8 and Cys283, of Acinetobacter soli L-asparaginase (AsA) were screened out via consensus design. After saturation mutagenesis and combinatorial mutation, the mutant C8Y/C283Q with highly improved thermostability was obtained with a half-life of 361.6 min at 40 °C, an over 34-fold increase compared with that of the wild-type. Its melting temperature (Tm) value reaches 62.3 °C, which is 7.1 °C higher than that of the wild-type. Molecular dynamics simulation and structure analysis revealed the formation of new hydrogen bonds of Gln283 and the aromatic interaction of Tyr8 formed with adjacent residues, resulting in enhanced thermostability. The improvement in the thermostability of L-asparaginase could efficiently enhance its effect on acrylamide inhibition; the contents of acrylamide in potato chips were efficiently reduced by 86.50% after a mutant C8Y/C283Q treatment, which was significantly higher than the 59.05% reduction after the AsA wild-type treatment. In addition, the investigation of the mechanism behind the enhanced thermostability of AsA could further direct the modification of L-asparaginases for expanding their clinical and industrial applications.

Prediction models for monitoring selenium and its associated heavy-metal accumulation in four kinds of agro-foods in seleniferous area.

Se-rich agro-foods are effective Se supplements for Se-deficient people, but the associated metals have potential risks to human health. Factors affecting the accumulation of Se and its associated metals in Se-rich agro-foods were obscure, and the prediction models for the accumulation of Se and its associated metals have not been established. In this study, 661 samples of Se-rich rice, garlic, black fungus, and eggs, four typical Se-rich agro-foods in China, and soil, matrix, feed, irrigation, and feeding water were collected and analyzed. The major associated metal for Se-rich rice and garlic was Cd, and that for Se-rich black fungus and egg was Cr. Se and its associated metal contents in Se-rich agro-foods were positively correlated with Se and metal contents in soil, matrix, feed, and matrix organic contents. The Se and Cd contents in Se-rich rice grain and garlic were positively and negatively correlated with soil pH, respectively. Eight models for predicting the content of Se and its main associated metals in Se-rich rice, garlic, black fungus, and eggs were established by multiple linear regression. The accuracy of the constructed models was further validated with blind samples. In summary, this study revealed the main associated metals, factors, and prediction models for Se and metal accumulation in four kinds of Se-rich agro-foods, thus helpful in producing high-quality and healthy Se-rich.

Enhancing the Catalytic Activity of Type II L-Asparaginase from Bacillus licheniformis through Semi-Rational Design.

Low catalytic activity is a key factor limiting the widespread application of type II L-asparaginase (ASNase) in the food and pharmaceutical industries. In this study, smart libraries were constructed by semi-rational design to improve the catalytic activity of type II ASNase from Bacillus licheniformis. Mutants with greatly enhanced catalytic efficiency were screened by saturation mutations and combinatorial mutations. A quintuple mutant ILRAC was ultimately obtained with specific activity of 841.62 IU/mg and kcat/Km of 537.15 min-1·mM-1, which were 4.24-fold and 6.32-fold more than those of wild-type ASNase. The highest specific activity and kcat/Km were firstly reported in type II ASNase from Bacillus licheniformis. Additionally, enhanced pH stability and superior thermostability were both achieved in mutant ILRAC. Meanwhile, structural alignment and molecular dynamic simulation demonstrated that high structure stability and strong substrate binding were beneficial for the improved thermal stability and enzymatic activity of mutant ILRAC. This is the first time that enzymatic activity of type II ASNase from Bacillus licheniformis has been enhanced by the semi-rational approach, and results provide new insights into enzymatic modification of L-asparaginase for industrial applications.

Solid-state fermentation with Rhizopus oligosporus RT-3 enhanced the nutritional properties of soybeans.

Fermented soybean products are favorite foods worldwide because of their nutritional value and health effects. In this study, solid-state fermentation (SSF) of soybeans with Rhizopus oligosporus RT-3 was performed to investigate its nutraceutical potential. A rich enzyme system was released during SSF. Proteins were effectively transformed into small peptides and amino acids. The small peptide content increased by 13.64 times after SSF for 60 h. The antioxidant activity of soybeans was enhanced due to the release of phenolic compounds. The soluble phenolic content increased from 2.55 to 9.28 gallic acid equivalent (GAE) mg/g after SSF for 60 h and exhibited high correlations with microbial enzyme activities during SSF. The potential metabolic pathways being triggered during SSF indicated that the improved nutritional composition of soybean attributed to the biochemical reactions catalyzed by microbial enzymes. These findings demonstrated that SSF could evidently improve the nutritional value and prebiotic potential of soybeans.

Transcriptome analysis provides new insight into the distribution and transport of selenium and its associated metals in selenium-rich rice.

Selenium is an essential trace element for humans and obtained from diary diets. The consumption of selenium-rich agricultural food is an efficient way to obtain selenium, but the quality and safety of selenium-rich agro-food are always affected by their associated heavy metals, even poses a potential threaten to human health. In this research, a sampling survey of heavy metals contents in selenium-rich rice was conducted, 182 sets of selenium-rich rice samples were collected from five selenium-rich rice-producing areas of China, and the accumulation of selenium and cadmium were found to be associated in rice and soil. Subsequently, a pot experiment was performed in the greenhouse via treating the soil samples with 12 different concentrations of selenium and heavy metals, and the contents of selenium and cadmium in rice grain were confirmed to be significantly associated. Moreover, transcriptome analysis revealed that the up-regulation of transporter-coding may promote the absorption of selenium and cadmium. The expression of antioxidant-coding genes and cadmium chelator transporter coding-genes was up-regulated to reduce the toxicity of cadmium. Meanwhile, the up-regulation of key genes of the ascorbic acid-glutathione metabolic pathway were responsible for the association between selenium and cadmium in Se-rich rice. Our work suggested the correlation between selenium and cadmium accumulation in selenium-rich rice, clarified their accumulation mechanism, provides a direction for the scientific production of selenium-rich agro-foods.

Characterization of a Novel L-Asparaginase from Mycobacterium gordonae with Acrylamide Mitigation Potential.

L-asparaginase (E.C.3.5.1.1) is a well-known agent that prevents the formation of acrylamide both in the food industry and against childhood acute lymphoblastic leukemia in clinical settings. The disadvantages of L-asparaginase, which restrict its industrial application, include its narrow range of pH stability and low thermostability. In this study, a novel L-asparaginase from Mycobacterium gordonae (GmASNase) was cloned and expressed in Escherichia coli BL21 (DE3). GmASNase was found to be a tetramer with a monomeric size of 32 kDa, sharing only 32% structural identity with Helicobacter pylori L-asparaginases in the Protein Data Bank database. The purified GmASNase had the highest specific activity of 486.65 IU mg-1 at pH 9.0 and 50 °C. In addition, GmASNase possessed superior properties in terms of stability at a wide pH range of 5.0-11.0 and activity at temperatures below 40 °C. Moreover, GmASNase displayed high substrate specificity towards L-asparagine with Km, kcat, and kcat/Km values of 6.025 mM, 11,864.71 min-1 and 1969.25 mM-1min-1, respectively. To evaluate its ability to mitigate acrylamide, GmASNase was used to treat potato chips prior to frying, where the acrylamide content decreased by 65.09% compared with the untreated control. These results suggest that GmASNase is a potential candidate for applications in the food industry.

Structures of l-asparaginase from Bacillus licheniformis Reveal an Essential Residue for its Substrate Stereoselectivity.

l-Asparaginase, which catalyzes the hydrolysis of l-asparagine, is an important enzyme in both the clinical and food industry. Exploration of efficient l-asparaginase with high substrate specificity, especially high chiral selectivity, is essential for extending its use. Herein, various crystal structures of type I l-asparaginase from Bacillus licheniformis (BlAsnase) have been resolved, and we found that there are two additional tyrosines in BlAsnase, contributing to the binding and catalysis of d-asparagine. Strikingly, the substitution of Tyr278 with methionine impaired the interaction with d-asparagine via water molecules due to the small hydrophobic side chain of methionine, which forced the ligand to the deep side of the active site toward the catalytic residues and thus resulted in the loss of hydrolyzing function. Our investigation of the substrate recognition mechanism of BlAsnase is significant for both a better understanding of l-asparaginase and its rational design to achieve high specificity for clinical and industrial applications.

Characterization of a novel type I l-asparaginase from Acinetobacter soli and its ability to inhibit acrylamide formation in potato chips.

l-Asparaginases have the potential to inhibit the formation of acrylamide, a harmful toxin formed during high temperature processing of food. A novel bacterium which produces l-asparaginase was screened. Type I l-asparaginase gene from Acinetobacter soli was cloned and expressed in Escherichia coli. The recombinant l-asparaginase had an activity of 42.0 IU mL-1 and showed no activity toward l-glutamine and d-asparagine. The recombinant l-asparaginase exhibited maximum catalytic activity at pH 8.0 and 40°C. The enzyme was stable in the pH ranging from 6.0 to 9.0. The activity of the recombinant enzyme was substantially enhanced by Ba2+, dithiothreitol, and ß-mercaptoethanol. The Km and Vmax values of the l-asparaginase for the l-asparagine were 3.22 mmol L-1 and 1.55 IU µg-1, respectively. Moreover, the recombinant l-asparaginase had the ability to mitigate acrylamide formation in potato chips. Compared with the untreated group, the content of acrylamide in samples treated with the enzyme was effectively decreased by 55.9%. These results indicate that the novel type I l-asparaginase has the potential for application in the food processing industry.

Improvement of the activity of l-asparaginase I improvement of the catalytic activity of l-asparaginase I from Bacillus megaterium H-1 by in vitro directed evolution.

Directed evolution methodologies have been used as promising strategies for improving the catalytic properties of many existing enzymes. In the presented work, this approach was applied to improve the enzyme activity of l-asparaginase I obtained from Bacillus megaterium H-1. After two rounds of error-prone polymerase chain reaction (epPCR) and two generations of sequential DNA shuffling, all of 5 different mutants showed a significant increase in the enzyme activity of l-asparaginase I, ranging from 6.27 to 22.78 IU/mL. Among these mutants, D-9B and DD-12G displayed the relatively high catalytic activity, which were 20.22-fold and 21.33-fold higher than the wild-type enzyme (WT), respectively. Furthermore, the catalytic efficiency (kcat/Km) of D-9B and DD-12G were also improved, which were 132.73 min-1mM-1 and 146.39 min-1mM-1, respectively, in comparison to that of WT (3.39 min-1mM-1). In addition, mutant DD-12G showed tolerance toward wider range of pH values and higher temperatures than its WT counterpart. Homology modeling of above two mutants reflected a reduction of hydrogen bonds and an introduction of flexible residues in the loops near the active catalytic site Thr15. These changes contributed to the flexibility of loops, which may lead to further enhancement in catalytic efficiency. Results also showed that approximately 88.5% (0.978 mg/kg) acrylamide could be removed from mutant DD-12G pre-treated fried potato chips. This study clearly shows that directional evolution methods can indeed be utilized to improve the activity of l-asparaginase, which could also provide research basis for future application in food industry.