The formation of carbon monoxide (CO) from glucose and cellulose by the treatment with various ionic liquids was studied. Ionic liquids with an imidazolium structure as cation and the chloride or acetate as anion were used. Additionally, 1,8-diazabicyclo [5.4.0]undec-7-ene (DBU) was employed as an additive. CO was generated from glucose with a maximum yield of 0.57mol% after 90min of treatment at 120°C in the reaction system in which DBU was added to the ionic liquid. Pyrolysis above 600°C has been commonly employed for the gasification of lignocellullosics to produce useful gases such as CO. However, this study has revealed that gasification of lignocellullosics to produce CO can occur at significantly lower temperature, specifically at 120°C.

Structure of biopolymers produced by microalgae plays an important role for their potential biological activity prediction and applications. Previously isolated and well characterized dominant fractions (Dch5-8) from ion-exchange chromatography separation of the biologically active microalga Dictyosphaerium chlorelloides exopolysaccharide (Dch) were pooled and partially acid hydrolyzed. The dominant sugar components in the combined Dch5-8 fraction were Gal and its 2-O-methyl derivative, Rha and Man, all accounting for about 94mol% of total amount of sugars. Separation of obtained hydrolysate on Bio-Gel P-2 afforded ten fractions. Their main components were identified by NMR. Based on oligosaccharide structures, the repeating unit of the polysaccharide backbone was identified as →2)-α-L-Rhap-(1→4)-2-O-methyl-[3-O-β-D-Galp]-α-D-Galp-(1→ branched by Man. Furthermore, the higher molecular weight fraction contained glucuronorhamnan. NMR data indicate 1,4-linked Rha units in the backbone in α and β configuration, branched at O2 by 2,4-di-O-methyl-β-d-glucuronic acid.

Most reported polysaccharides from Poria cocos (PCPs) in traditional Chinese medicine decoctions were water-soluble heteropolysaccharides while the water-insoluble PCPs were scarcely researched due to the poor water-solubility. In this study, a water-insoluble polysaccharide with high yield of 59%, and high purity with a glucan content of 98.8%, was isolated by diluted sodium hydroxide at low temperature and coded as PCPA. The chemical structure of PCPA was identified as a liner β-glucan with 1, 3-linked glycosidic bond by the fourier infrared spectrum (FT-IR), ion chromatography (ICP), gas chromatography and mass spectrometry (GC-MS), and nuclear magnetic resonance (NMR) measurements. Importantly, PCPA was successfully used to construct hydrogels (PCPA-Gs) with good thermal stability, water retention ability and swelling property through simple physical cross-linking, due to the abundance of hydroxyl groups on glucan chains. Moreover, the rheology analysis of PCPA-Gs showed a rapid transition between gel and sol as well as the shear-thinning property. The hydrogel developed in this study holds promise for applications in the food, pharmaceutical, and cosmetic fields.

A reusable electrochemical glassy carbon electrode (GCE) platform based on the acid-responsive host-guest interaction between β-cyclodextrin (β-CD) and benzimidazole (BM) derivatives was developed. The β-CD can specifically recognize the BM derivative through the acid -responsive host-guest interaction. The electrode was first modified by eletrografting to immobilize a diamine linker (Boc-EDA), resulting in GCEBoc-EDA in which one amine was used for covalent immobilization to the electrode and another Boc protected amine was used to solid-phase synthesis on following step-by-step modifications on the electrode. After deprotection of the Boc group on the GCEBoc-EDA, carbonyldiimidazole (CDI)-activated β-CD was coupled with -NH2 on the electrode to result in GCEβ-CD. Due to the nonspecific interaction, we further improved the GCEβ-CD electrode by introducing immobilized poly(ethylene glycol) methyl ether (PEG-Me) to result in GCEβ-CD/PEG-Me, along with optimized procedures. CV, DPV, and EIS methods were applied for recording the electrochemistry signals. We utilized GCEβ-CD/PEG-Me to investigate the host-guest interaction and found the electrochemical signal exhibited dynamic behavior. The GCEβ-CD/PEG-Me was able to regenerate the β-CD surface more than 20 times after HCl acidic washes. We further investigated the interaction of carbendazim (CBZ), a commonly used fungicide in the agriculture and food industry, and observed a positive electrochemical response. The sensor design has potential applications in ensuring food safety.

The production of biofuels from lignocellulosic biomass using carbohydrate-active enzymes like cellulases is key to a sustainable energy production. Understanding the adsorption mechanism of cellulases and associated binding domain proteins down to the molecular level details will help in the rational design of improved cellulases. In nature, carbohydrate-binding modules (CBMs) from families 17 and 28 often appear in tandem appended to the C-terminus of several endocellulases. Both CBMs are known to bind to the amorphous regions of cellulose non-competitively and show similar binding affinity towards soluble cello-oligosaccharides. Based on the available crystal structures, these CBMs may display a uni-directional binding preference towards cello-oligosaccharides (based on how the oligosaccharide was bound within the CBM binding cleft). However, molecular dynamics (MD) simulations have indicated no such clear preference. Considering that most soluble oligosaccharides are not always an ideal substrate surrogate to study the binding of CBMs to the native cell wall or cell surface displayed glycans, it is critical to use alternative reagents or substrates. To better understand the binding of type B CBMs towards smaller cello-oligosaccharides, we have developed a simple solid-state depletion or pull-down binding assay. Here, we specifically orient azido-labeled carbohydrates from the reducing end to alkyne-labeled micron-sized bead surfaces, using click chemistry, to mimic insoluble cell wall surface-displayed glycans. Our results reveal that both family 17 and 28 CBMs displayed a similar binding affinity towards cellohexaose-modified beads, but not cellopentaose-modified beads, which helps rationalize previously reported crystal structure and MD data. This may indicate a preferred uni-directional binding of specific CBMs and could explain their co-evolution as tandem constructs appended to endocellulases to increase amorphous cellulose substrate targeting efficiency. Overall, our proposed workflow can be easily translated to measure the affinity of glycan-binding proteins to click-chemistry based immobilized surface-displayed carbohydrates or antigens.

N-Glycosyltransferase (NGT) is an inverting glycosyltransferase for an unusual pathway of N-linked protein glycosylation and glycosylates polypeptides in the consensus sequon (N-(X≠P)-T/S) with hexose monosaccharides. Here, we expressed and characterized a novel N-glycosyltransferase from Mannheimia haemolytica (named MhNGT). RP-HPLC and Mass Spectrometry were used to assay and quantify glycopeptide formation by MhNGT and determine its substrate specificities. MhNGT could utilize a variety of nucleotide-activated sugar donors, including UDP-Glc, UDP-Gal and UDP-Xyl, to glycosylate the tested peptides DANYTK, GGNWTT and PAVGNCSSALR with higher efficiency than ApNGT which was comprehensive studied. The optimum temperature of MhNGT was about 30°C and the optimum pH was 7.5-8.0 in PBS-NaOH buffer. MhNGT exhibited a different position-specific residue preference of substrate peptides from the NGT of Actinobacillus pleuropneumoniae (ApNGT). The effective glycosylation of common short peptides by MhNGT demonstrated the enzyme's potential to alter therapeutically significant mammalian N-glycoproteins.

Oxidation of β-cyclodextrin (β-CD) using varying molar ratios of sodium periodate (NaIO4) was investigated in detail on synthesis, characterization and antibacterial property. Synthesis and characterization results showed that Oxidized β-cyclodextrins (OX-β-CDs) were obtained and aldehyde (CHO) groups were successfully introduced. Our results demonstrated that aldehyde content and yield increased with increasing NaIO4 molar amount. However, the structure of β-CD was degraded as a result of glycosidic ring opening with increasing stoichiometric ratio of NaIO4/β-CD to 5/1 and 7/1. Aldehyde functional groups in OX-β-CDs were characterized by employing FTIR, 1H NMR, XRD, SEM techniques and confirmed by the detection of CHO peak at 1730 cm-1 in the FTIR and detection of the aldehyde H peak between 9 to 10ppm in the 1H NMR spectrum. In addition, SEM and XRD of OX-β-CDs showed alterations in the morphological and crystal structure (transforming from crystalline to amorphous) of β-CD as a result of increasing oxidation. Especially, antibacterial activity of OX-β-CDs was investigated against both Gram-negative and Gram-positive bacteria by using the minimal inhibitory concentration (MIC) and the Disk diffusion method. The results showed that OX-β-CDs possessed good antibacterial activity, which can destroy the bacterial cell wall, and may be used as an antibacterial agent.

The discovery of new glycosylation reactions is still a major challenge in carbohydrate chemistry. Traditional glycosylation reactions require the preparation of sugar donors with anomeric active or latent leaving groups. Dehydrative glycosylation is a fascinating alternative that enables the direct formation of the glycosidic bond from the hemiacetal, eliminating the need for (sometimes unstable) leaving groups, and allowing to reduce reaction, work-up, and purification times. Although some interesting methods of dehydrative glycosylation have been reported, in order to compete with conventional chemical glycosylation, a greater number of efficient and stereoselective methods need to be developed. Herein, a dehydrative procedure that uses a combination of iodine, triphenylphosphine, and a base (DMAP or imidazole) is described. This methodology allows for the preparation of sugar derivatives from commercially available 1-hydroxy glycosyl donors. The reaction takes place under mild conditions through the in situ-formation of an anomeric iodide intermediate, which, upon reaction with an alcohol, gives the corresponding glycosides up to quantitative yields and with high α-stereoselectivity.