Preparation and functional properties of rice bran globulin-chitooligosaccharide-quercetin-resveratrol covalent complex.

BACKGROUND: As the major protein (approximately 36%) in rice bran, globulin exhibits excellent foaming and emulsifying properties, endowing its useful application as a foaming and emulsifying agent in the food industry. However, the low water solubility restricts its commercial potential in industrial applications. The present study aimed to improve this protein's processing and functional properties. RESULTS: A novel covalent complex was fabricated by a combination of the Maillard reaction and alkaline oxidation using rice bran globulin (RBG), chitooligosaccharide (C), quercetin (Que) and resveratrol (Res). The Maillard reaction improved the solubility, emulsifying and foaming properties of RBG. The resultant glycosylated protein was covalently bonded with quercetin and resveratrol to form a (RBG-C)-Que-Res complex. (RBG-C)-Que-Res exhibited higher thermal stability and antioxidant ability than the native protein, binary globulin-chitooligosaccharide or ternary globulin-chitooligosaccharide-polyphenol (only containing quercetin or resveratrol) conjugates. (RBG-C)-Que-Res exerted better cytoprotection against the generation of malondialdehyde and reactive oxygen species in HepG2 cells, which was associated with increased activities of antioxidative enzymes superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) through upregulated genes SOD1, CAT, GPX1 (i.e. gene for glutathione peroxidase-1), GCLM (i.e. gene for glutamate cysteine ligase modifier subunit), SLC1A11 (i.e. gene for solute carrier family 7, member 11) and SRXN1 (i.e. gene for sulfiredoxin-1). The anti-apoptotic effect of (RBG-C)-Que-Res was confirmed by the downregulation of caspase-3 and p53 and the upregulation of B-cell lymphoma-2 gene expression. CONCLUSION: The present study highlights the potential of (RBG-C)-Que-Res conjugates as functional ingredients in healthy foods. © 2024 Society of Chemical Industry.

Systems metabolic engineering upgrades Corynebacterium glutamicum to high-efficiency cis, cis-muconic acid production from lignin-based aromatics.

Lignin displays a highly challenging renewable. To date, massive amounts of lignin, generated in lignocellulosic processing facilities, are for the most part merely burned due to lacking value-added alternatives. Aromatic lignin monomers of recognized relevance are in particular vanillin, and to a lesser extent vanillate, because they are accessible at high yield from softwood-lignin using industrially operated alkaline oxidative depolymerization. Here, we metabolically engineered C. glutamicum towards cis, cis-muconate (MA) production from these key aromatics. Starting from the previously created catechol-based producer C. glutamicum MA-2, systems metabolic engineering first discovered an unspecific aromatic aldehyde reductase that formed aromatic alcohols from vanillin, protocatechualdehyde, and p- hydroxybenzaldehyde, and was responsible for the conversion up to 57% of vanillin into vanillyl alcohol. The alcohol was not re-consumed by the microbe later, posing a strong drawback on the producer. The identification and subsequent elimination of the encoding fudC gene completely abolished vanillyl alcohol formation. Second, the initially weak flux through the native vanillin and vanillate metabolism was enhanced up to 2.9-fold by implementing synthetic pathway modules. Third, the most efficient protocatechuate decarboxylase AroY for conversion of the midstream pathway intermediate protocatechuate into catechol was identified out of several variants in native and codon optimized form and expressed together with the respective helper proteins. Fourth, the streamlined modules were all genomically combined which yielded the final strain MA-9. MA-9 produced bio-based MA from vanillin, vanillate, and seven structurally related aromatics at maximum selectivity. In addition, MA production from softwood-based vanillin, obtained through alkaline depolymerization, was demonstrated.

Traditional or adaptive design of experiments? A pilot-scale comparison on wood delignification.

Traditional design of experiments and response surface methodology are widely used in engineering and process development. Bayesian optimization is an alternative machine learning approach that adaptively selects successive experimental conditions based on a predefined performance measure. Here we compared the two approaches using simulations and empirical experiments on alkaline wood delignification to identify important benefits and drawbacks of Bayesian optimization in the context of design of experiments. The simulations showed that the selection of initial experiments and measurement noise influenced the convergence of the Bayesian optimization algorithm to known optimal conditions. Both methods, however, showed comparable pilot-scale results on optimal digestion conditions, where high cellulose yields were combined with acceptable kappa numbers and pulp viscosities. Bayesian optimization did not enable a decrease in the number of experiments required for reaching these conditions but provided a more accurate model in the vicinity of the optimum based on additional modelling and cross-validation. These results shed light on the practical differences between the two methodologies for process development and are an important contribution to the chemometrics and machine learning communities.

Stepwise enzymatic hydrolysis of alkaline oxidation treated sugarcane bagasse for the co-production of functional xylo-oligosaccharides and fermentable sugars.

High chemical input is required for enzymatic production of xylo-oligosaccharides (XOS) using xylan extracted from lignocellulosic biomass. In this study, enzymatic hydrolysis of alkaline oxidation (AO) treated sugarcane bagasse (SCB) directly for the production of XOS was conducted. The effect of AO pretreatment on the chemical compositions and hydrolytic properties of SCB was investigated. The AO pretreatment conditions with low chemical input for the production of XOS were optimized by orthogonal design. Stepwise enzymatic hydrolysis of AO pretreated SCB with xylanase and cellulase produced XOS (1.78 g/L), meanwhile, the cellulose conversion increased from 84.97% to 91.51% compared with directly enzymatic hydrolysis using cellulase. HPLC-UV and MALDI-TOF-MS analysis indicated that the obtained XOS products were mainly composed of xylobiose and xylose with a small amount of arabinose/4-O-methylglucuronic acid substituted xylotriose and xylotetraose. The proposed strategy for the co-production of functional XOS and fermentable sugars from SCB showed potential of industrial application.

Improving the enzymatic hydrolysis of larch by coupling water pre-extraction with alkaline hydrogen peroxide post-treatment and adding enzyme cocktail.

Soluble arabinogalactan (AG) in larch leads to reagent waste during its biorefining using oxidative pretreatment strategies. A two-stage pretreatment of water pre-extraction followed by alkaline hydrogen peroxide (AHP) pretreatment was investigated to more efficiently convert larch cellulose into glucose, while also obtaining a value-added AG product stream. The results showed that water pre-extraction increases the lignin selectivity of both NaOH and H2O2 reagents, translating to improved lignin removal and enzymatic hydrolysis yields. This was found to be related to cellulose accessibility alongside the effective consumption of the reagents. Moreover, the addition of mannanase also significantly enhanced enzymatic digestibility of pretreated solid from 81.0% to 97.7% (4% H2O2 charge and 180 °C) when 40 U/g mannanase was supplemented with 20 FPU/g cellulase. In all, it was demonstrated that coupling mannanase with cellulase could improve larch's enzymatic digestibility and overall viability for biorefining processes.

Degradation of Vanillin During Lignin Valorization Under Alkaline Oxidation.

The preparation of vanillin from lignin is one of the lignin valorization strategies. However, obtaining high vanillin yield is still a challenge. Therefore, the process of vanillin production and factors that affect yield of vanillin has attracted much attention. Here, oxidation of vanillin was performed to study its degradation behavior under lignin alkaline oxidation conditions. High-performance liquid chromatography, liquid chromatography-electrospray mass spectrometry, gas chromatography-mass spectrometer and gel permeation chromatography were employed to analyze the products including monomers and dimers. Results demonstrated that reaction temperature and time greatly affected vanillin degradation; vanillin can be completely converted in 5 h at 160 °C. At 160 °C, the main products of vanillin oxidation were small molecule acids and alcohols, other monophenols, and even condensed dimers. A possible vanillin degradation pathway was proposed. The results indicate that vanillin degradation and condensation are the main reasons for decreasing vanillin yield during lignin valorization under alkaline oxidation circumstances.