Lignin from Annual Plants as Raw Material Source for Flavors and Basic Chemicals

The enzymatic conversion of lignins, possibly in combination with electrochemical oxidation, makes aromatics such as syringol, guaiacol, vanillin and catechol available in the qualities required by the fragrance industry. The lignins were obtained by soda digestion from wheat straw and Miscanthus, characterized and then converted with laccases. The overall yield amounted up to 9 wt % with a product spectrum confined to four substances. Catechol was the major product, with a fraction of ≈75 %. It can easily be isolated by extraction with acetone.     

Insights into biological delignification of rice straw by Trametes hirsuta and Myrothecium roridum and comparison of saccharification yields with dilute acid pretreatment

Rice straw is the most abundant agricultural residue on a global scale and is widely available as feedstock for cellulosic fuel production. However, it is highly recalcitrant to biochemical deconstruction and also generates inhibitors that affect enzymatic saccharification. Rice straw from eastern Arkansas was subjected to dilute acid pretreatment (160 °C, 48 min and 1.0% sulfuric acid) and solid-state fermentation with two lignocellulolytic fungi, Trametes hirsuta and Myrothecium roridum, and their saccharification efficacies were compared. T. hirsuta and M. roridum were tested separately; pretreatment of rice straw with either strain for seven days resulted in 19 and 70% enrichment of its holocellulose content, respectively. However, liquid chromatography analysis of the alkali extracts showed significant differences in cell wall degradation by T. hirsuta and M. roridum. T. hirsuta removed 15% more phenolic compounds and 38% more glucan than M. roridum, while M. roridum removed 77% more xylan than T. hirsuta. Fungal and dilute acid pretreated biomass was then hydrolyzed using Accellerase^® 1500, a saccharification cocktail. Saccharification efficiency of M. roridum was 37% higher than that of dilute acid pretreatment of rice straw, requiring 8% lower enzyme loading and 50% shorter enzymatic hydrolysis duration. Alkali extraction of fungal pretreated biomass also yielded 10–15 g kg⁻¹ of acid precipitable polymeric lignin (APPL), which is a valuable co-product for biorefineries. In comparison to dilute acid pretreatment, fungal pretreatment could be a cost-effective alternative for the degradation of recalcitrant biomass, such as rice straw.     

Rheological behaviour of processed avocado pulp emulsions

Shear rate vs. shear stress data were obtained on avocado pulp in water emulsions using a concentric cylinder rheometer and fitted to a power law model. Dilution, as volume fraction of water, had a pronounced effect on the apparent viscosity of the pulp emulsions and the Richardson equation, (η_R= exp.( a φ) for the emulsion viscosity fitted the data well. A mean slope coefficient, a , of 4.57 can be used as a first approximation. Enzymatic treatment (40°C, 1 h), is slightly more effective than thermal treatment (65°C, 1 h), in reducing the initial apparent viscosity of the pulp-water emulsions     

Chemospecific ring-opening polymerization of α-methylenemacrolides

α-Methylenemacrolides having various groups, such as aromatic, ether, and amine, were enzymatically, anionically, and radically polymerized. The polymerization with the lipase catalyst successfully afforded polymers only through the ring-opening process, whereas the vinyl polymerizations selectively proceeded by using anionic and radical initiators. The polyesters obtained by the enzymatic polymerization have a polymerizable methacrylic methylene group in the main-chain, in addition to the aromatic and polar groups, and were further radically polymerized to quantitatively produce a cross-linked polymer gel.     

AMMONIA CATALYZED ORGANOSOLV DELIGNIFICATION OF POPLAR

A parametric investigation of NH₄OH catalyzed solvent delignification of poplar was conducted to define pretreatment conditions which would yield an optimal separation of the biomass components and an enzymatic susceptible solid carbohydrate phase. Delignification parameters of interest included concentration of NH₄OH, time and temperature of the reaction, and type of solvent. The addition of 0.82 M NH₄OH to the delignification liquor increased lignin removal and decreased carbohydrate degradation, but increasing NH₄OH concentration had no additional effect. At lower reaction temperatures, the extent of delignification increased with reaction time; at higher temperatures, a “relignification” of the pretreated wood was observed. The delignification and hemicellulose solubilization were modelled and rate constants reported. No major difference between three potential pulping solvents—ethanol, butanol, phenol—was observed. The enzymatic susceptibility of pretreated wood samples was approximately 6-fold greater than that of the untreated poplar. UV absorbance was used to qualitatively characterize the soiubilized lignins.     

Synthesis of polymeric prodrugs of chlorphenesin with saccharide branches by chemo-enzymatic regioselective strategy

A facile and regioselective enzymatic synthesis approach to prepare polymerizable lipophilic chlorphenesin vinyl esters was developed in this research. The influence of different organic solvents, enzyme sources, reaction time and the acylation reagent on the synthesis of chlorphenesin vinyl esters was investigated. Then the polymerizable monomers 1-O-vinylsuccinyl-chlorphenesin (OVSC) and 1-O-vinyladipoyl-chlorphenesin (OVAC) were homopolymerized using AIBN as the initiator. The obtained polymeric prodrugs were characterized with IR, NMR and GPC analyzes. The poly-OVSC has M_n of 1.35 × 10⁴ and M_w/M_n of 1.95, and the poly-OVAC has M_n of 2.37 × 10⁴ and M_w/M_n of 4.30. Moreover, 6-O-vinyladipoyl-d-glucose (OVAG), a biocompatible monomer, was copolymerized with OVSC and OVAC. Polymeric prodrugs of chlorphenesin with saccharide branches were successfully obtained with high molecular weight.     

Effect of particle size on quality of crab meatballs using enzymatically deproteinized crab by-products

The production of crab meatballs generates large amount of crab shell waste, it is therefore necessary to develop a green, economical and environmentally friendly process to vaporize the waste. This study was aimed at investigating the applicability of microwave heating combined with ultrasonic field-assisted alkaline protease (MUSED) (50 ℃, pH = 9.0, 14025.67 U/g, 4.21 h and liquid/material = 14.41:1) for pretreatment of ball-milled crab shells. The ball milling efficiency of the crab shell powder pretreated by MUSED was observed to increase by 50 % compared to the control group, with the final average particle size of D₄ = 4.88 ± 0.20 um. High calcium solubility and low energy consumption of the ball-milled powder increased dietary calcium bioavailability and reduced the potential for high calorie intake. The addition of 6 % (w/w) crab shell powder treated by MUSED improved the texture of the crab meatballs (CM-D₄) and gave the product enhanced crab flavor relative to the control group. Moreover, the lower cooking loss of CM-D₄ resulted in significant retention of nutrients (p < 0.05). The crab shell powder treated by MUSED method was more efficient in ball milling, which improved the quality of crab meatballs while relieving environmental pressure.     

Hybrid phenol biosensor based on modified phenoloxidase electrode

Electrolytic deposition has been widely used to immobilize biomacromolecules, and it is always the most important factor to preserve or even increase an activity of the immobilized protein. We report here simple and rather universal method for the highly efficient immobilization of laccase for amperometric biosensing. Laccase from Cerrena unicolor has been successfully immobilized (electrolytic deposition) on the surface of thin, ordered polythiophene films (3-methylthiophene/3-thiopheneacetic acid/N-heptyl-3,6-bis(2-thiophene)carbazole). Two different compounds capable of mediating laccase-catalyzed reactions have been tested by cyclic voltammetry. They exhibited quasi-reversible electrodic behaviour with formal redox potentials ranging from 68 and 918 mV (E⁰vs. SCE). The immersion of the laccase-coated electrode in solution with substrate generated large catalytic currents easily recorded by cyclic voltammetry at low potential scan rates. Considering the fact, that immobilization strategy showed high efficiency, obtained results suggest that method for phenoloxidase immobilization has a great potential of enabling high throughput fabrication of bioelectronics’ devices.     

Design optimization of an enzymatic assay in an electrokinetically-driven microfluidic device

Microfluidic systems are increasingly popular for rapid and cheap determinations of enzyme assays and other biochemical analysis. In this study reduced order models (ROM) were developed for the optimization of enzymatic assays performed in a microchip. The model enzyme assay used was β-galactosidase (β-Gal) that catalyzes the conversion of Resorufin β-d-galactopyranoside (RBG) to a fluorescent product as previously reported by Hadd et al. (Anal Chem 69(17): 3407–3412, 1997). The assay was implemented in a microfluidic device as a continuous flow system controlled electrokinetically and with a fluorescence detection device. The results from ROM agreed well with both computational fluid dynamic (CFD) simulations and experimental values. While the CFD model allowed for assessment of local transport phenomena, the CPU time was significantly reduced by the ROM approach. The operational parameters of the assay were optimized using the validated ROM to significantly reduce the amount of reagents consumed and the total biochip assay time. After optimization the analysis time would be reduced from 20 to 5.25 min which would also resulted in 50% reduction in reagent consumption.