A high-performance size-exclusion chromatography (HPSEC) method coupled to Evaporative Light Scattering (ELS) and Refractive Index (RI) detectors were evaluated and compared for the molecular mass (Mw) estimation of pectin in a wide range (0.342-805 kDa). Instrumental parameters of the ELSD were optimised by Response Surface Methodology (RSM) being 73 °C the evaporator temperature and 0.9 mL/min the air flow rate. The linear range for the ELSD concentration response was wider (10-2250 mg/L) and better (R = 0.985) than RID (10-1500 mg/L; R = 0.875). The limits of detection (LOD) and quantitation (LOQ) for all pullulans hardly changed in ELSD (LOD: 1.22-1.99 mg/L; LOQ: 4.07-6.63 mg/L); however, RID showed huge variations (LOD: 0.49-10.41 mg/L; LOQ: 1.64-34.70 mg/L), which increased with the Mw. In general, responses of both detectors were similar for the Mw estimation, although pectin characterisation with HPSEC-ELSD exhibited better results in the lowest Mw compounds.
The intensification of processes is essential for the sustainability of the biorefinery concept. Enzyme catalysis assisted by ultrasound (US) may offer interesting opportunities in the agri-food sector because the cavitation effect provided by this technology has been shown to improve the efficiency of the biocatalysts. This review presents the recent advances in this field, focused on three main applications: ultrasound-assisted enzymatic extractions (UAEE), US hydrolysis reactions, and synthesis reactions assisted by US for the manufacturing of agri-food produce and ingredients, enabling the upgrading of agro-industrial waste. Some theoretical and experimental aspects of US that must be considered are also reviewed. Ultrasonic intensity (UI) is the main parameter affecting the catalytic activity of enzymes, but a lack of standardization for its quantification makes it unsuitable to properly compare results. Applications of enzyme catalysis assisted by US in agri-foods have been mostly concentrated in UAEE of bioactive compounds. In second place, US hydrolysis reactions have been applied for juice and beverage manufacturing, with some interesting applications for producing bioactive peptides. In last place, a few efforts have been performed regarding synthesis reactions, mainly through trans and esterification to produce structured lipids and sugar esters, while incipient applications for the synthesis of oligosaccharides show promising results. In most cases, US has improved the reaction yield, but much information is lacking on how different sonication conditions affect kinetic parameters. Future research should be performed under a multidisciplinary approach for better comprehension of a very complex phenomenon that occurs in very short time periods.
An optimization of temperature, time, and extracting agent concentration of pectin extraction from sunflower heads using sodium citrate and nitric acid (SP-SC and SP-NA) was carried out. At optimal conditions, the yield of extraction with nitric acid (SPO-NA) was twofold greater than the corresponding with sodium citrate (SPO-SC) (14.3 versus 7.7%, respectively). Regarding pectin structure, the galacturonic acid (GalA) content in both, SPO-SC and SPO-NA, was similar (∼85%). However, SPO-NA showed lower molecular weight (Mw) (88.9 kDa) and neutral sugar content (4%) than SPO-SC (464 kDa, 9%), indicating that nitric acid deeply degraded pectin structure. These differences derived into dissimilar behavior in their technological functionality. SPO-SC showed higher viscosity and better emulsifying capacity than SPO-NA, although any of them were able to stabilize the oil/water emulsion. Both sunflower pectins formed gels with Ca (75 mg/g of pectin) at pH 3.0. However, when sucrose was added, the gels formed by SP-SC and 20% sucrose presented the same hardness as those of SP-NA with 40% sucrose. These results suggest that the pectin extracted with sodium citrate, an eco-friendly agent, could be a promising ingredient, with good thickening and gelling properties.
Galactooligosaccharides (GOS), recognised prebiotic, can be industrially produced from lactose and commercial β-galactosidase (β-gal) from Kluyveromyces lactis. Residual lactose and glucose limit GOS applications. To handle this problem, a multienzymatic system, with β-gal and glucose oxidase (Gox), was proposed to reduce glucose content in reaction media through its oxidation to gluconic acid (GA). Besides, ultrasound (US) probe effect over the multienzymatic system to produce GOS and GA has been evaluated. A production around 40% of GOS was found in all treatments after the first hour of reaction. However, glucose consumption and GA production was significantly higher (P < 0.05) for sequential reaction assisted by US, obtaining the best production of GOS (49%) and GA (28%) after 2 h of reaction. The conformational and residual activity changes of enzymes under US conditions were also evaluated, Gox being positively affected whereas in β-gal hardly any change was found.
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