Search

This research is focused on improving performance of chitosan based functional material by introducing active additive. A series of assays revealed incorporation with clove essential oil (CEO) significantly improved the physical, chemical and antimicrobial performance of chitosan. In this work, the prepared chitosan-CEO film (CH-CEO) showed varieties in color parameters, mechanical strength and water vapor permeability. Moreover, chitosan was endowed with significant antioxidant and antimicrobial activities, thereby it was used as protective coatings for fresh-cutting apple tubes at ~1 °C. Results demonstrated the treatment slowed down the quality deterioration process of preserved apple samples, especially for firmness and color. As well, CH-CEO coating reduced microbial counts in the preserved apple samples and inhibited the varieties in the chemical properties. The overall observations revealed that CH-CEO film has remarkable potential as an antioxidant and antimicrobial material, especially as an active coating for fresh-cutting foods during storage.

Enzymatic digestion of sodium alginate to produce specific oligosaccharides has attracted great attention. However, commercial enzymes that efficiently produce specific oligosaccharides are still unavailable. In the present study, a novel gene encoding an alginate lyase (designated alg7A) was cloned from the marine bacterium Vibrio sp. W13 and expressed in E. coli. The recombinant Alg7A shows high activities toward alginate, poly-α-l-guluronate (polyG), poly-β-d-mannuronate (polyM) and polyMG, and more preferred to polyMG. Moreover, the enzyme contains a highly conserved domain of the Polysaccharide Lyase (PL) 7 family (R*E*R, Q*H and Y*KAG*Y*Q), which indicates that it belongs to PL7. Furthermore, the thin layer chromatography and ESI-MS analysis showed that Alg7A mainly releases trisaccharides from alginate. These results demonstrated that Alg7A has a great potential to be used to produce oligosaccharides from alginate.

To investigate the effect and mechanism of chitosan nanoparticles (CSNPs) on the germination and seedling growth of wheat (Triticum aestivum L.), we conducted systematic research on the impact of different concentrations (1–100 μg/mL) of CSNPs and chitosan (CS). The result of energy-dispersive spectroscopy (EDS) and confocal laser scanning microscopy (CLSM) showed that adsorption of CSNPs on the surface of wheat seeds was higher than that of CS. CSNPs had growth promoting effect at a lower concentration (5 μg/mL) compared with CS (50 μg/mL). In addition, the application of 5 μg/mL CSNPs induced the auxin-related gene expression, accelerated indole-3-acetic acid (IAA) biosynthesis and transport, and reduced IAA oxidase activity resulting in the increase of IAA concentration in wheat shoots and roots. The results suggest that CSNPs have positive effect on seed germination and seedling growth of wheat at a lower concentration than CS due to higher adsorption on the surface of wheat seeds.

Oligogalacturonides (OGAs) are a biologically active carbohydrate derived from homogalacturonan, a major element of cell wall pectin. OGAs induced resistance and mechanism were assessed in Arabidopsis thaliana-Pseudomonas syringae pv. tomato DC3000 (Pst DC3000) interaction. The effective resistance was mainly observed at 25 mg/L OGAs with reduced disease index, bacterial multiplication, higher transcript level of salicylic acid (SA) pathway related genes (PR1, PR2, PR5) and jasmonic acid (JA) pathway related genes (PDF1.2, VSP2) as well as SA, JA content and production of reactive oxygen species (ROS), nitric oxide (NO). In SA (NahG, sid2) and JA (jar1) deficient mutants, disease severity indicated that both SA and JA pathways are necessary for Arabidopsis response to Pst DC3000. OGAs triggered less resistance to Pst DC3000 in JA-deficient mutant, and SA-deficient mutants signifying that SA and JA play redundant roles in OGAs induced resistance. Therefore, these evidences further reveal the signaling pathways of OGAs resistance, which is conducive to its application in agriculture to protect plants from diseases.

RNA-binding proteins (RBPs) are pivotal participants in post-transcriptional gene regulation. They interact with RNA directly to perform several post-transcriptional RNA regulatory functions or direct metabolic processes. Despite the essential importance, the understanding of plant RBPs is elementary, which derives mainly from other kingdoms via bioinformatic extrapolation or mRNA-binding proteins captured through UV crosslinked method. Recently, orthogonal organic phase separation (OOPS) method for RBP identification has been used in mammals and Escherichia coli. And plentiful RBPs were enriched without molecular tagging or capture of polyadenylated RNA in an unbiased way. In our study, OOPS was conducted on Arabidopsis and 468 RBPs were discovered including 244 putative RBPs. There were 17 peroxidases in 232 RBPs with enzymatic activities. In addition, Arabidopsis thaliana-Pst DC3000-chitinpentaose interaction system was chosen to explore whether OOPS can be used to dig specific RBPs under special physiological conditions. Eighty-four differential RBPs in this system were found and some of them involved in reactive oxygen species (ROS) metabolic pathway. These results showed OOPS can be applied to plants successfully and would be a useful method to identify RBPomes and specific RBPs.

A new oligoalginate lyase encoding gene, designed oal17A, was cloned from marine bacterium Vibrio sp. W13, and then expressed in Escherichia coli. The recombinant Oal17A was purified by NTA-Ni resin with maximal activity at 30°C and pH7.0. Oal17A exhibited broad substrate specificity, and preferred to degrade alginate than polyM or polyG into monosaccharide acid. The specific activity of Oal17A toward alginate, polyM and polyG was 21.14U/mg, 12.31U/mg and 7.43U/mg, respectively. With features of high-level expression and broad substrate specificity, Oal17A would be a potential tool for alginate monomer production process of alginate utilizing for biofuels and bioethanol production.

In order to produce highly purified (1→3)-β-d-glucan oligosaccharides ((1→3)-β-d-GOS) monomers, a hydrophilic interaction liquid chromatography (HILIC) system with X-Amide stationary phase was performed. Nine (1→3)-β-d-GOS monomers with degree of polymerization (DP) from 2 to 10 were successfully separated. Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) demonstrated that these monomers were with high purity. Furthermore, a hydrogen peroxide (H2O2) online detection method was established to monitor H2O2 releases in tobacco cells. This is the first report on nine consecutive (1→3)-β-d-GOS monomers purification and its effect upon H2O2-releasing in plants. It was found that (1→3)-β-d-GOS monomers with higher DP induced stronger defense responses in plants, which will pave the way for elucidating the relationship between (1→3)-β-d-GOS and biological activities.

A new chitosanase gene of glycoside hydrolase (GH) family 75, csnw2, was cloned from an isolated strain Aspergillus sp. W-2 (CGMCC7018). The mature CsnW2 protein, fused to His-tag at C-terminus, was expressed in Pichia pastoris X-33 and purified with the affinity chromatography of Ni2+-NTA. The novel recombinant CsnW2 showed maximal activity with chitosan at pH 6.0 and 55 degrees C. Moreover, it had good pH stability and thermostability at a broad pH range of 3.0-10.0 and a temperature range of 30-70 degrees C, respectively. The enzymatic activity of the CsnW2 could be significantly enhanced by Ca2+, Mn2+ and Mg2+ at a concentration of 1 mM, but strongly inhibited by Fe2+, Zn2+, Ge2+, Ni2+ and Cu2+ above 1 mM. The CsnW2 showed specific hydrolytic activity against chitosan and preferred to hydrolyze chitosan with high degree of deacetylation. The main products of chtiosan (92% deacetylation) were chitosan oligosaccharides (COS) with degree of polymerization (DP) arranging from 2 to 6. Combined with the hydrolysis of COS from DP2 to DP6, CsnW2 was considered to be an endo-acting chitosanase. (C) 2015 Elsevier B.V. All rights reserved.