Demei Meng

Methyl Salicylate-Induced Arginine Catabolism Is Associated with Up-regulation of Polyamine and Nitric Oxide Levels and Improves Chilling Tolerance in Cherry Tomato Fruit

The effects of methyl salicylate (MeSA) on chilling injury (CI) and gene expression levels, enzyme activities, and metabolites related to arginine catabolism in cherry tomato fruit were investigated. Freshly harvested fruits were treated with 0.05 mM MeSA vapor at 20 °C for 12 h and then stored at 2 °C for up to 28 days. MeSA reduced CI and enhanced the accumulation of putrescine, spermidine, and spermine, which was associated with increased gene expression levels and activities of arginase, arginine decarboxylase, and ornithine decarboxylase at most sampling times. MeSA also increased nitric oxide synthase activity, which at least partly contributed to the increased nitric oxide content. The results indicate that MeSA activates the different pathways of arginine catabolism in cold-stored fruit and that the reduction in CI by MeSA may be due to the coordinated metabolism of arginine and the increase in polyamines and nitric oxide levels.

Simultaneous Decoloration and Deproteinization of Crude Polysaccharide from Pumpkin Residues by Cross-Linked Polystyrene Macroporous Resin

A novel method for the purification of crude polysaccharide from fermentation broth of pumpkin residues by macroporous resins was developed. Through static adsorption and desorption and adsorption kinetic tests, six resins (AB-8, S-8, HPH480, HPD100, X-5, and D101) with different polarity, diameter, and surface area were studied for simultaneous decoloration and deproteinization of crude polysaccharide, and S-8 was chosen as the best one. Dynamic breakthrough and desorption tests were performed in a glass column packed with S-8 resin, and the resulting adsorption ratios of pigment and protein were 84.3% and 75.9% (w/w), respectively, with a recovery ratio of polysaccharide 84.7% (w/w). S-8 resin also exhibited higher purification efficiency than the other tested traditional methods. Moreover, UV/vis spectroscopy (200-900 nm) analysis revealed most of the pigment and protein were absorbed by S-8 resin, and HPLC (containing a refractive index detector and a HPSEC column) results indicated that there was no degradation of the polysaccharide. This automated and efficient method via adsorption-desorption strategy could have potential in scale-up purification and preparation of polysaccharide in the future.

Postharvest Application of Methyl Jasmonate for Improving Quality Retention of Agaricus bisporus Fruit Bodies

The influence of methyl jasmonate (MeJA) on postharvest quality and enzyme activities, gene expression level, and the functional component content linked to postharvest deterioration in Agaricus bisporus (J.E. Lange) Imbach fruit bodies was investigated. Freshly harvested fruit bodies were treated with 0 (control), 10 and 100 μM MeJA vapor at 20 °C for 12 h and then stored at 10 °C for up to 7 days. The results indicated that treatments with 100 μM MeJA vapor maintained a high level of soluble protein and total sugar, delayed browning, promoted the accumulation of phenolics and flavonoids, and inhibited the increase of respiratory rate and membrane leakage. Furthermore, 100 μM MeJA inhibited the activities of polyphenoloxidase, increased the antioxidant enzymes activities of catalase and superoxide dismutase, and lowered relative expression levels of three genes encoding polyphenol oxidase (AbPPO1, AbPPO2, and AbPPO3) throughout the storage period. Comparatively, 10 μM MeJA also had a clear beneficial effect on postharvest mushroom quality maintenance but was not as effective as 100 μM MeJA treatment. These findings suggest that application of MeJA could have potential in maintaining the quality of harvested A. bisporus fruit bodies.

Novel reassortant influenza viruses between pandemic (H1N1) 2009 and other influenza viruses pose a risk to public health.

Influenza A virus (IAV) is characterized by eight single-stranded, negative sense RNA segments, which allows for gene reassortment among different IAV subtypes when they co-infect a single host cell simultaneously. Genetic reassortment is an important way to favor the evolution of influenza virus. Novel reassortant virus may pose a pandemic among humans. In history, three human pandemic influenza viruses were caused by genetic reassortment between avian, human and swine influenza viruses. Since 2009, pandemic (H1N1) 2009 (pdm/09 H1N1) influenza virus composed of two swine influenza virus genes highlighted the genetic reassortment again. Due to wide host species and high transmission of the pdm/09 H1N1 influenza virus, many different avian, human or swine influenza virus subtypes may reassert with it to generate novel reassortant viruses, which may result in a next pandemic among humans. So, it is necessary to understand the potential threat of current reassortant viruses between the pdm/09 H1N1 and other influenza viruses to public health. This study summarized the status of the reassortant viruses between the pdm/09 H1N1 and other influenza viruses of different species origins in natural and experimental conditions. The aim of this summarization is to facilitate us to further understand the potential threats of novel reassortant influenza viruses to public health and to make effective prevention and control strategies for these pathogens.

Hot air treatment-induced arginine catabolism is associated with elevated polyamines and proline levels and alleviates chilling injury in postharvest tomato fruit

BACKGROUND To understand whether arginine catabolism might be involved in hot air (HA)-induced chilling tolerance mechanism in tomato fruit, we investigated the effect of HA treatment on endogenous arginine catabolism in relation to chilling injury. RESULTS Tomato fruit were harvested at mature green stage and treated with HA at 38°C for 12 h and then stored at 2°C for 21 days. The effects of HA treatment on fruit chilling injury and gene expression levels or enzyme activity, and metabolites related to arginine catabolism were evaluated. HA treatment reduced the chilling injury symptoms of tomato fruit and enhanced the accumulation of endogenous polyamines, especially putrescine and proline. This accumulation is associated with the increased transcript levels of genes encoding arginase (LeARG1 and LeARG2), arginine decarboxylase (LeADC), ornithine decarboxylase (LeODC) and ornithine aminotransferase (LeOAT) at most sampling times. However, HA treatment had little effect on nitric oxide synthase activity and nitric oxide concentration. CONCLUSION These results revealed that the reduction in chilling injury by HA treatment may be due to the accumulation of putrescine and proline induced primarily by activating the catabolism of endogenous arginine.

Saccharification of pumpkin residues by coculturing of Trichoderma reesei RUT-C30 and Phanerochaete chrysosporium Burdsall with delayed inoculation timing.

Trichoderma reesei and Phanerochaete chrysosporium with different lignocellulose-degrading enzyme systems have received much attention due to their ability to biodegrade lignocellulosic biomass. However, the synergistic effect of the two fungi on lignocellulose degradation is unknown. Herein, a cocultivation of T. reesei RUT-C30 and P. chrysosporium Burdsall for biodegradation of lignocellulosic pumpkin residues (PRS) was developed to produce soluble saccharide. Results indicated that a cocultivation of the two fungi with P. chrysosporium Burdsall inoculation delayed for 1.5 days produced the highest saccharide yield of 53.08% (w/w), and only 20.83% (w/w) of PRS were left after one batch of fermentation. In addition, this strategy increased the activities of secreted cellulases (endoglucanase, cellobiohydrolase, and β-glucosidase) and ligninases (lignin peroxidase and manganese peroxidase), which correlated to the increased saccharide yield. Besides, the resulting monosaccharides including glucose (1.23 mg/mL), xylose (0.13 mg/mL), arabinose (0.46 mg/mL), and fructose (0.21 mg/mL) from cocultures exhibited much higher yields than those from monoculture, which provides basal information for further fermentation research. This bioconversion of PRS into soluble sugars by cocultured fungal species provides a low cost method based on lignocellulose for potential biofuels or other bioproduct production.

Synthesis and immunogenicity of PG-tb1 monovalent glycoconjugate

A PG-tb1 hapten from the West Beijing strains of Mycobacterium tuberculosis cell wall has been efficiently synthesized and conjugated to CRM197 in a simple way as linker-equipped carbohydrate by applying squaric acid chemistry for an original neoglycoprotein, creating a potent T-dependent conjugate vaccine. The intermediate monoester can be easily purified and the degree of incorporation can be monitored by MALDI-TOF mass spectrometry. After administered systemically in mice without any adjuvant, the conjugate induced high antigen-specific IgG levels in serum. Furthermore, following the third immunization, significant antibody titers frequently exceeding 0.8 million were observed in the sera of mice vaccinated with PG-CRM197 conjugate which showed the potential for preparation of TB vaccine.

Synergistic Effects of l-Arginine and Methyl Salicylate on Alleviating Postharvest Disease Caused by Botrysis cinerea in Tomato Fruit

The effects of l-arginine (Arg, 1 mM) and/or methyl salicylate (MeSA, 0.05 mM) treatment on gray mold caused by Botrytis cinerea in tomato fruit were studied. Results indicated that Arg or MeSA alleviated the incidence and severity of fruit disease caused by B. cinerea, and that both Arg and MeSA (Arg + MeSA) further inhibited the development of fruit decay. Treatment with Arg + MeSA not only enhanced the activities of superoxide dismutase, catalase, and peroxidase but also promoted the expression levels of pathogenesis-related protein 1 gene and the activities of defense-related enzymes of phenylalanine ammonia-lyase, polyphenol oxidase, β-1,3-glucanase, and chitinase during most of the storage periods, which were associated with lower disease incidence and disease index. In addition, the combined treatment elevated the levels of total phenolics, polyamines, especially putrescine, and nitric oxide. These observations suggest that treatment of fruit with Arg + MeSA is an effective and promising way to alleviate postharvest decays on a commercial scale.

Identification and active site analysis of the 1-aminocyclopropane-1-carboxylic acid oxidase catalysing the synthesis of ethylene in Agaricus bisporus.

BACKGROUND 1-Aminocyclopropane-1-carboxylate oxidase (ACO) is a key enzyme that catalyses the final step in the biosynthesis of the plant hormone ethylene. Recently, the first ACO homologue gene was isolated in Agaricus bisporus, whereas information concerning the nature of the ethylene-forming activity of this mushroom ACO is currently lacking. METHODS Recombinant ACO from A. bisporus (Ab-ACO) was purified and characterised for the first time. Molecular modelling combined with site-directed mutagenesis and kinetic and spectral analysis were used to investigate the property of Ab-ACO. RESULTS Ab-ACO has eight amino acid residues that are conserved in the Fe (II) ascorbate family of dioxygenases, including four catalytic residues in the active site, but Ab-ACO lacks a key residue, S289. In comparison to plant ACOs, Ab-ACO requires ACC and Fe (II) but does not require ascorbate. In addition, Ab-ACO had relatively low activity and was completely dependent on bicarbonate, which could be ascribed to the replacement of S289 by G289. Moreover, the ferrous ion could induce a change in the tertiary, but not the secondary, structure of Ab-ACO. CONCLUSIONS These results provide crucial experimental support for the ability of Ab-ACO to catalyse ethylene formation in a similar manner to that of plant ACOs, but there are differences between the biochemical and catalytic characteristics of Ab-ACO and plant ACOs. GENERAL SIGNIFICANCE This work enhances the understanding of the ethylene biosynthesis pathways in fungi and could promote profound physiological research of the role of ethylene in the regulation of mushroom growth and development.

Urea-Driven Epigallocatechin Gallate (EGCG) Permeation into the Ferritin Cage, an Innovative Method for Fabrication of Protein–Polyphenol Co-assemblies

The 8 nm diameter cavity endows the ferritin cage with a natural space to encapsulate food components. In this work, urea was explored as a novel medium to facilitate the formation of ferritin-polyphenol co-assemblies. Results indicated that urea (20 mM) could expand the 4-fold channel size of apo-red bean ferritin (apoRBF) with an increased initial iron release rate υ0 (0.22 ± 0.02 μM min-1) and decreased α-helix content (5.6%). Moreover, urea (20 mM) could facilitate the permeation of EGCG into the apoRBF without destroying the ferritin structure and thus form ferritin-EGCG co-assemblies (FECs) with an encapsulation ratio and loading capacity of 17.6 and 2.1% (w/w), respectively. TEM exhibited that FECs maintained a spherical morphology with a 12 nm diameter in size. Fluorescence analysis showed that urea intervention could improve the binding constant K [(1.22 ± 0.8) × 104 M-1] of EGCG to apoRBF. Furthermore, the EGCG thermal stability was significantly improved (20-60 °C) compared with free EGCG. Additionally, this urea-involved method was applicable for chlorogenic acid and anthocyanin encapsulation by the apoRBF cage. Thus, urea shows potential as a novel potential medium to encapsulate and stabilize bioactive polyphenols for food usage based on the ferritin protein cage structure.