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1.
Carbohydr Polym ; 343: 122467, 2024 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-39174128

RESUMEN

Individual cells are the smallest units of the plant tissue structure, and their structure and physicochemical properties are essential for whole food processing. In this study, cassava cells were isolated using acid-alkali, hydrothermal, and pectinase methods, and the differences in microstructure and physicochemical properties among the cells, starch, and whole flour were investigated. Cassava cells isolated using pectinase showed intact individual cells with a higher isolation rate and less damage to the cell wall structure and intracellular composition. The presence of cell walls in intact individual cells inhibited the swelling and leaching of starch, resulting in a lower peak viscosity and higher gelatinization temperature than those of starch. The intact cell structure and non-starch composition enhanced the shear resistance of the gels in the sample. Heat treatment led to the gelatinization of intracellular starch and increased the permeability of the cell wall, destroying the physical barrier function of the cell wall; however, the compact cell matrix and non-starch components can inhibit starch hydrolysis. This study suggests that cells isolated using pectinase can be used to investigate the effect of cell walls on the functional properties of intracellular starch in cassava. The isolated cells provide new insights into the cassava industry.


Asunto(s)
Pared Celular , Harina , Manihot , Almidón , Manihot/química , Almidón/química , Harina/análisis , Pared Celular/química , Viscosidad , Poligalacturonasa/metabolismo , Poligalacturonasa/química , Hidrólisis
2.
PeerJ ; 12: e17715, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-39119104

RESUMEN

Postharvest rot caused by various fungal pathogens is a damaging disease affecting kiwifruit production and quality, resulting in significant annual economic losses. This study focused on isolating the strain P3-1W, identified as Diaporthe eres, as the causal agent of 'Hongyang' postharvest rot disease in China. The investigation highlighted cell wall degrading enzymes (CWDEs) as crucial pathogenic factors. Specially, the enzymatic activities of cellulase, ß-galactosidase, polygalacturonase, and pectin methylesterases peaked significantly on the second day after infection of D. eres P3-1W. To gain a comprehensive understanding of these CWDEs, the genome of this strain was sequenced using PacBio and Illumina sequencing technologies. The analysis revealed that the genome of D. eres P3-1W spans 58,489,835 bp, with an N50 of 5,939,879 bp and a GC content of 50.7%. A total of 15,407 total protein-coding genes (PCGs) were predicted and functionally annotated. Notably, 857 carbohydrate-active enzymes (CAZymes) were identified in D. eres P3-1W, with 521 CWDEs consisting of 374 glycoside hydrolases (GHs), 108 carbohydrate esterase (CEs) and 91 polysaccharide lyases (PLs). Additionally, 221 auxiliary activities (AAs), 91 glycosyltransferases (GTs), and 108 carbohydrate binding modules (CBMs) were detected. These findings offer valuable insights into the CAZymes of D. eres P3-1W.


Asunto(s)
Actinidia , Ascomicetos , Genoma Fúngico , Enfermedades de las Plantas , Actinidia/microbiología , Enfermedades de las Plantas/microbiología , China , Ascomicetos/genética , Ascomicetos/patogenicidad , Ascomicetos/enzimología , Genoma Fúngico/genética , Poligalacturonasa/genética , Poligalacturonasa/metabolismo , Frutas/microbiología , Hidrolasas de Éster Carboxílico/genética , Hidrolasas de Éster Carboxílico/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Celulasa/genética , Celulasa/metabolismo , Pared Celular/metabolismo , beta-Galactosidasa/genética , beta-Galactosidasa/metabolismo
3.
Curr Microbiol ; 81(8): 255, 2024 Jul 02.
Artículo en Inglés | MEDLINE | ID: mdl-38955830

RESUMEN

Turkey litter waste is lignocellulosic and keratinous, requiring prior enzymatic treatment to facilitate fiber hydrolysis and utilization by microorganisms in anaerobic digestion (AD) process. The understanding of the performance of microorganisms in AD can be facilitated through molecular biology and bioinformatics tools. This study aimed to determine the taxonomic profile and functional prediction of microbial communities in the AD of turkey litter waste subjected to enzymatic pretreatment and correlate it with operational parameters. The tests involved the use of turkey litter (T) at 25 g L-1 of volatile solids, a granular inoculum (S) (10% m/v), and the addition of cellulase (C), and pectinase (P) enzymes at four concentrations. The use of enzymes increased methane production by 19% (turkey litter, inoculum, and cellulase-TSC4) and 15% (turkey litter, inoculum, and enzymatic pectinase-TSP4) compared to the control (turkey litter and inoculum-TS), being more effective in TSC4 (667.52 mLCH4), where there was consumption of acetic, butyric, and propionic acids. The pectinase assay (TSP4) showed a methane production of 648 mLCH4 and there was the accumulation of metabolites. Cellulolytic microorganisms Bacteroides, Ruminofilibacter, Lachnospiraceae, Ruminococcaceae, and Methanosaeta were favored in TSC4. In TSP4, the predominant genus was Macellibacteroides and Methanosarcina, and genes involved in methylotrophic methanogenesis were also found (mtaB, mtmB, and mtbB). Enzymes involved in hydrogenotrophic methanogenesis were identified in both assays (TSC4 and TSP4). Molecular tools helped to understand the metabolic routes involved in AD with enzymatic treatment, allowing the elaboration of strategies to improve the sustainable degradation of turkey litter waste.


Asunto(s)
Bacterias , Celulasa , Metano , Poligalacturonasa , Pavos , Anaerobiosis , Animales , Metano/metabolismo , Celulasa/metabolismo , Bacterias/clasificación , Bacterias/genética , Bacterias/metabolismo , Bacterias/aislamiento & purificación , Pavos/microbiología , Poligalacturonasa/metabolismo , Hidrólisis , Lignina/metabolismo , Agricultura , Metagenómica
4.
Ultrason Sonochem ; 107: 106933, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-38865900

RESUMEN

Extraction of anthocyanins from Lycium ruthenicum Murr. (L. ruthenicum) is a notable challenge in food production, requiring methods that balance efficiency and safety. In this study, we conducted a comparative analysis the extraction of anthocyanins by natural air drying (NAD), vacuum freeze drying (VFD), hot air drying (HAD), and vacuum microwave drying (MVD) combined with ultrasonic-assisted enzymolysis extraction (UAEE). The results demonstrated that the extraction yield and antioxidant activity of anthocyanins were significantly higher in VFD. This phenomenon can be attributed to the modification of raw material's microstructure, leading to an increased extraction yield of specific anthocyanins such as Cyanidin-3-galactoside, Delphinidin chloride, Cyanidin, and Petunidin. According to the pretreatment results, the extraction process of anthocyanins was further optimized. The highest yield (3.16 g/100 g) was obtained in following conditions: 0.24 % pectinase, 48 °C, solid:liquid = 1:21, and 21 min ultrasonic time. This study improves the commercial value and potential application of L. ruthenicum in food industry.


Asunto(s)
Antocianinas , Desecación , Lycium , Antocianinas/aislamiento & purificación , Antocianinas/química , Lycium/química , Desecación/métodos , Ondas Ultrasónicas , Fraccionamiento Químico/métodos , Antioxidantes/aislamiento & purificación , Antioxidantes/química , Poligalacturonasa , Microondas
5.
Int J Biol Macromol ; 274(Pt 2): 133416, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-38925202

RESUMEN

Bioaugmentation retting with the specialized pectinolytic and xylanolytic microorganisms can accelerate the removal of non-cellulosic macromolecules around plant fibers, thus shortening retting time and facilitating fiber quality. Currently, few specialized microorganisms have been explored for the retting of sisal fibers. The present study excavated the retting fungi including Aspergillus micronesiensis HD 3-6, Penicillium citrinum HD 3-12-3, and Cladosporium sp. HD 4-13 from the region-specific soil samples of planting sisal, and investigated their bioaugmentation retting effects on raw sisal leaves. Results showed that combination of the three fungi achieved the most excellent degumming efficiency (13.69 % of residual gum in sisal fibers) and the highest fiber yield (4.47 %). Furthermore, this fungi combination had the ideal enzymatic hydrolysis features with high activities of pectinase, xylanase and mannanase whereas a low activity of cellulase during the whole retting process, thus endowing the prepared sisal fibers with the lowest mass percentage of non-cellulosic macromolecules (9.76 wt%) and the highest cellulose content (89.23 wt%). SEM and FT-IR analysis further verified that the non-cellulosic substances around sisal fibers were efficiently removed. In summary, the consortia of the three fungi achieved ideal degumming-related enzymes for the removal of non-cellulosic macromolecules, thus acquiring the efficient preparation of sisal fibers.


Asunto(s)
Sasa , Sasa/química , Sasa/microbiología , Hidrólisis , Hongos , Celulosa/química , Celulosa/metabolismo , Poligalacturonasa/metabolismo , Penicillium/enzimología , Sustancias Macromoleculares/química , Sustancias Macromoleculares/metabolismo
6.
Ultrason Sonochem ; 108: 106948, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-38878717

RESUMEN

In this study, the process conditions, physicochemical properties, structural composition and activity of polysaccharides isolated from leechee peel (LPP) by ultrasound-assisted extraction (UAE) with enzyme and alkali solution extraction (ASE) were compared. The results showed that the total sugar content of LPP extracted by UAE accounted for 75.65 %, which was significantly higher than that extracted by alkali solution. The optimum conditions were as follows: extraction temperature of 68.78 ℃, ultrasonic enzymolysis time of 39.68 min, pectinase dosage of 4.03 %, solid-liquid ratio of 1:30 g/mL, and ultrasonic power of 360 W. The antioxidant activities and structure of leechee peel polysaccharide (LPP) prepared under different conditions were compared. It was found that UAE-LPP was an α-type polysaccharide containing 15.83 % uronic acid. Moreover, LPP extracted by UAE showed strong activity in anti-lipid peroxidation and reducing ability. Ultrasound-assisted enzymatic method is an effective means to improve the content and activity of natural plant polysaccharides, and this method has the advantages of short time-consuming, simple process and easy operation, which can greatly improve the utilization rate of polysaccharides and lay a theoretical and scientific basis for the development and utilization of LPP.


Asunto(s)
Antioxidantes , Fraccionamiento Químico , Polisacáridos , Ondas Ultrasónicas , Polisacáridos/química , Polisacáridos/aislamiento & purificación , Fraccionamiento Químico/métodos , Antioxidantes/química , Antioxidantes/aislamiento & purificación , Poligalacturonasa , Temperatura , Peroxidación de Lípido/efectos de los fármacos
7.
Ultrason Sonochem ; 108: 106955, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-38909597

RESUMEN

The main purpose of the present study was to determine the effect of associating an optimized ultrasound-assisted extraction (UAE) protocol with enzyme-assisted extraction (EAE) in aqueous media, using the dried berries of Hippophae rhamnoides L. (sea buckthorn) as plant material. A specialized software was used for the determination of potential optimal extraction parameters, leading to the development of four optimized extracts with different characteristics (UAE ± EAE). For these extracts, buffered or non-buffered solutions have been used, with the aim to determine the influence of adjustable pH on extractability. As enzymatic solution, a pectinase, cellulase, and hemicellulase mix (2:1:1) has been applied, acting as pre-treatment for the optimized protocol. The highest extractive yields have been identified for non-buffered extracts, and the E-UAE combination obtained extracts with the highest overall in vitro antioxidant activity. The HPLC-MSn analysis demonstrated a rich composition in different types of isorhamnetin-O-glycosides, as well as some quercetin-O-glycosides, showing a high recovery of specific flavonol-type polyphenolic species. Moreover, we have tentatively identified two flavanols (i.e., catechin and epigallocatechin) and one flavone derivative (i.e., luteolin).


Asunto(s)
Fraccionamiento Químico , Flavonoides , Frutas , Glicósidos , Hippophae , Ondas Ultrasónicas , Hippophae/química , Glicósidos/química , Glicósidos/aislamiento & purificación , Frutas/química , Flavonoides/aislamiento & purificación , Flavonoides/química , Flavonoides/análisis , Fraccionamiento Químico/métodos , Agua/química , Poligalacturonasa/química , Poligalacturonasa/metabolismo , Antioxidantes/química , Antioxidantes/aislamiento & purificación , Glicósido Hidrolasas/metabolismo , Celulasa/metabolismo , Desecación/métodos , Concentración de Iones de Hidrógeno
8.
J Oleo Sci ; 73(7): 963-976, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38945925

RESUMEN

The objective of this research was to evaluate the efficiency of aqueous enzymatic extraction (AEE) to obtain oil from hemp seeds (Cannabis sativa L.) grown in northern Morocco. Optimisation of AEE extraction parameters, including pH, enzyme concentration (hemicellulase, protease and pectinase), temperature and incubation time, to maximize oil yield was achieved using response surface methodology with a central composite design. For comparison, the solvent extraction (Soxhlet) (SE) method was also used. Optimized hydrolysis conditions involved incubation for 4 hours at 60°C with a pH of 6.5, using a multi-enzyme preparation comprising protease, hemicellulase and pectinase at concentrations of 55, 202.5 and 234 U/mg, respectively. Referring to the conventional Soxhlet extraction (SE), Aqueous Enzymatic Extraction (AEE) achieved a 30.65% oil recovery rate under the optimized parameters mentioned above. The use of enzymes produced an oil that was more stable against oxidation than the solvent-extracted oil, with a peroxide value (PV) of 19.54 and 47.87 meq O 2 /kg, respectively. Furthermore, HPLC-DAD analysis of tocopherol content indicated a higher total tocopherol content (547.2 mg/kg) in Aqueous Enzymatic Extraction (AEE) compared to Soxhlet Extraction (SE) (513.51 mg/kg), with γ-tocopherol being the predominant form. No significant differences in fatty acid composition were observed between the two extraction methods with linoleic acid and alpha-linolenic acid being the predominant constituents.


Asunto(s)
Cannabis , Glicósido Hidrolasas , Péptido Hidrolasas , Aceites de Plantas , Poligalacturonasa , Semillas , Cannabis/química , Poligalacturonasa/metabolismo , Aceites de Plantas/química , Aceites de Plantas/aislamiento & purificación , Glicósido Hidrolasas/metabolismo , Semillas/química , Péptido Hidrolasas/metabolismo , Hidrólisis , Extracción Líquido-Líquido/métodos , Calidad de los Alimentos , Agua , Tocoferoles/análisis , Tocoferoles/aislamiento & purificación , Concentración de Iones de Hidrógeno , Temperatura , Solventes/química , Tecnología Química Verde/métodos
9.
Arch Microbiol ; 206(7): 291, 2024 Jun 07.
Artículo en Inglés | MEDLINE | ID: mdl-38849576

RESUMEN

Biomass-degrading enzymes produced by microorganisms have a great potential in the processing of agricultural wastes. In order to produce suitable biomass-degrading enzymes for releasing sugars and aroma compounds from tobacco scraps, the feasibility of directly using the scraps as a carbon source for enzyme production was investigated in this study. By comparative studies of ten fungal strains isolated from tobacco leaves, Aspergillus brunneoviolaceus Ab-10 was found to produce an efficient enzyme mixture for the saccharification of tobacco scraps. Proteomic analysis identified a set of plant biomass-degrading enzymes in the enzyme mixture, including amylases, hemicellulases, cellulases and pectinases. At a substrate concentration of 100 g/L and enzyme dosage of 4 mg/g, glucose of 17.6 g/L was produced from tobacco scraps using the crude enzyme produced by A. brunneoviolaceus Ab-10. In addition, the contents of 23 volatile molecules, including the aroma compounds 4-ketoisophorone and benzyl alcohol, were significantly increased after the enzymatic treatment. The results provide a strategy for valorization of tobacco waste by integrating the production of biomass-degrading enzymes into the tobacco scrap processing system.


Asunto(s)
Aspergillus , Biomasa , Nicotiana , Nicotiana/microbiología , Nicotiana/metabolismo , Aspergillus/enzimología , Aspergillus/metabolismo , Azúcares/metabolismo , Odorantes/análisis , Proteínas Fúngicas/metabolismo , Glicósido Hidrolasas/metabolismo , Amilasas/metabolismo , Compuestos Orgánicos Volátiles/metabolismo , Hojas de la Planta/microbiología , Celulasas/metabolismo , Poligalacturonasa/metabolismo
10.
Environ Sci Pollut Res Int ; 31(27): 39217-39231, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38814560

RESUMEN

To overcome the human and animal survivability risk, sustainable development is the only option on earth that can be achieved through the maximum use of renewable environmental resources. Recycling of waste paper is an emerging waste management approach to conserve natural resources. Herein, we studied enzyme-mediated process to recycle the xerographic paper by using the crude fungal extract from indigenously isolated fungi identified as Aspergillus assiutensis. The fungal enzyme cocktail has been characterized for the production of multiple enzymes namely cellulase, amylase, xylanase, pectinase, and protease. All these enzymes have pH optima in the acidic range and except cellulase and all the enzymes are stable from 10 to 80 C. In the zymogram analysis, pectinase, xylanase, amylase, and cellulase were detected at 68 kDa, ~ 54 kDa, 38 kDa, and 30 kDa, respectively. Also, the presence of protease was confirmed by the clear zone at 68, 31, and 16 kDa. A 26% decrease in the kappa number and reduction in Hex A of the pulp was observed on the treatment of the pulp with enzyme as compared to the control pulp without any treatment. The physical and chemical properties of the pulp were also improved by enzyme-mediated pulping as compared to the control The physiochemical parameter of the effluent like TDS was reduced (397 ppm) significantly in comparison to chemical deinking process and it was within the permissible limit. BOD and alkalinity were reduced when the enzymes and chemical dosage were used in combination. These results indicate that chemi-enzymatic deinking is most promising to reduce or remove the pollution parameters including ink and this approach can be used in the paper and pulp industry for sustainable development.


Asunto(s)
Aspergillus , Papel , Reciclaje , Aspergillus/enzimología , Poligalacturonasa , Celulasa
11.
Int J Biol Macromol ; 269(Pt 2): 131918, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38697418

RESUMEN

Polygalacturonases (PGs) can modulate chemistry and mechanical properties of the plant cell wall through the degradation of pectins, one of its major constituents. PGs are largely used in food, beverage, textile, and paper industries to increase processes' performances. To improve the use of PGs, knowledge of their biochemical, structural and functional features is of prime importance. Our study aims at characterizing SmoPG1, a polygalacturonase from Selaginella moellendorffii, that belongs to the lycophytes. Transcription data showed that SmoPG1 was mainly expressed in S. moellendorffii shoots while phylogenetic analyses suggested that SmoPG1 is an exo-PG, which was confirmed by the biochemical characterization following its expression in heterologous system. Indeed, LC-MS/MS oligoprofiling using various pectic substrates identified galacturonic acid (GalA) as the main hydrolysis product. We found that SmoPG1 was most active on polygalacturonic acid (PGA) at pH 5, and that its activity could be modulated by different cations (Ca2+, Cu2+, Fe2+, Mg2+, Mn2+, Na2+, Zn2+). In addition, SmoPG1 was inhibited by green tea catechins, including (-)-epigallocatechin-3-gallate (EGCG). Docking analyses and MD simulations showed in detail amino acids responsible for the SmoPG1-EGCG interaction. Considering its expression yield and activity, SmoPG1 appears as a prime candidate for the industrial production of GalA.


Asunto(s)
Pectinas , Poligalacturonasa , Selaginellaceae , Poligalacturonasa/metabolismo , Poligalacturonasa/química , Poligalacturonasa/genética , Selaginellaceae/química , Selaginellaceae/genética , Selaginellaceae/enzimología , Pectinas/metabolismo , Pectinas/química , Filogenia , Especificidad por Sustrato , Simulación del Acoplamiento Molecular , Secuencia de Aminoácidos , Concentración de Iones de Hidrógeno , Hidrólisis , Ácidos Hexurónicos
12.
Int J Biol Macromol ; 269(Pt 2): 132054, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38704063

RESUMEN

In this study, we analyzed the pectin structure within the pulp of cassava. Cassava pectin, derived from cassava pulp treatment at 120 °C for 90 min, was separated into four fractions (CP-P, CP-SD1, CP-SD2F, and CP-SD2R) based on variations in water solubility, electrical properties, and molecular weights. Sugar composition analysis demonstrated an abundance of homogalacturonan (HG) in CP-P and CP-SD2F, rhamnogalacturonan I (RG-I) in CP-SD2R, and neutral sugars in CP-SD1. Because RG-I possesses a complex structure, we analyzed CP-SD2R using various pectinolytic enzymes. Galactose was the major sugar in CP-SD2R accounting for 49 %, of which 65 % originated from arabinogalactan I, 9 % from galactose and galactooligosaccharides, 5 % from arabinogalactan II, and 11 % from galactoarabinan. Seventy-four percent of arabinose in CP-SD2R was present as galactoarabinan. The methylation (DM) and acetylation (DAc) degrees of cassava pectin were 11 and 15 %, respectively. The HG and RG-I regions exhibited DAc values of 5 and 44 %, respectively, signifying the high DAc of RG-I compared to HG. Information derived from the structural analysis of cassava pectin will enable efficient degradation of pectin and cellulose, leading to the use of cassava pulp as a raw material for biorefineries.


Asunto(s)
Manihot , Pectinas , Manihot/química , Pectinas/química , Fraccionamiento Químico , Peso Molecular , Poligalacturonasa/química , Poligalacturonasa/metabolismo , Metilación , Solubilidad
13.
Biopolymers ; 115(4): e23586, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-38747448

RESUMEN

Cellulose nanofibers, a sustainable and promising material with widespread applications, exhibit appreciable strength and excellent mechanical and physicochemical properties. The preparation of cellulosic nanofibers from food or agricultural residue is not sustainable. Therefore, this study was designed to use three halophytic plants (Cressa cretica, Phragmites karka, and Suaeda fruticosa) to extract cellulose for the subsequent conversion to cellulosic nanofibers composites. The other extracted biomass components including lignin, hemicellulose, and pectin were also utilized to obtain industrially valuable enzymes. The maximum pectinase (31.56 IU mL-1), xylanase (35.21 IU mL-1), and laccase (15.89 IU mL-1) were produced after the fermentation of extracted pectin, hemicellulose, and lignin from S. fruticosa, P. karka, and C. cretica, respectively. Cellulose was methylated (with a degree of substitution of 2.4) and subsequently converted into a composite using polyvinyl alcohol. Scanning electron microscopy and Fourier-transform infrared spectroscopy confirmed the successful synthesis of the composites. The composites made up of cellulose from C. cretica and S. fruticosa had a high tensile strength (21.5 and 15.2 MPa) and low biodegradability (47.58% and 44.56%, respectively) after dumping for 3 months in soil, as compared with the composite from P. karka (98.79% biodegradability and 4.9 MPa tensile strength). Moreover, all the composites exhibited antibacterial activity against gram-negative bacteria (Escherichia coli and Klebsiella pneumoniae) and gram-positive bacteria (Staphylococcus aureus). Hence, this study emphasizes the possibility for various industrial applications of biomass from halophytic plants.


Asunto(s)
Celulosa , Celulosa/química , Plantas Tolerantes a la Sal/química , Plantas Tolerantes a la Sal/metabolismo , Lignina/química , Resistencia a la Tracción , Antibacterianos/farmacología , Antibacterianos/química , Antibacterianos/aislamiento & purificación , Poligalacturonasa/metabolismo , Poligalacturonasa/química , Espectroscopía Infrarroja por Transformada de Fourier , Lacasa/metabolismo , Lacasa/química , Nanofibras/química , Pectinas/química , Pectinas/aislamiento & purificación , Pectinas/metabolismo , Chenopodiaceae/química , Chenopodiaceae/metabolismo , Polisacáridos/química , Polisacáridos/aislamiento & purificación , Endo-1,4-beta Xilanasas/metabolismo , Endo-1,4-beta Xilanasas/química
14.
Int J Biol Macromol ; 266(Pt 2): 131309, 2024 May.
Artículo en Inglés | MEDLINE | ID: mdl-38580019

RESUMEN

Enzymatic degradation of plant biomass requires the coordinated action of various enzymes. In this study, the production of reducing sugars from pectic substrates and sugar beet pulp (SBP) was investigated and compared using commercial enzyme preparations, including M2, pectinase (E1), Viscozyme L (V-L) and L-40. V-L, a cellulolytic enzyme mix produced by Aspergillus sp. was further evaluated as the most robust enzyme cocktail with the strongest SBP degradation ability in terms of the release of monosaccharides, methanol, and acetate from SBP. Mass-spectrometry-based proteomics analysis of V-L revealed 156 individual proteins. Of these, 101 proteins were annotated as containing a carbohydrate-active enzyme module. Notably, of the 50 most abundant proteins, ca. 44 % were predicted to be involved in pectin degradation. To reveal the role of individual putative key enzymes in pectic substrate decomposition, two abundant galacturonases (PglA and PglB), were heterologously expressed in Pichia pastoris and further characterized. PglA and PglB demonstrated maximum activity at 57 °C and 68 °C, respectively, and exhibited endo-type cleavage patterns towards polygalacturonic acid. Further studies along this line may lead to a better understanding of efficient SBP degradation and may help to design improved artificial enzyme mixtures with lower complexity for future application in biotechnology.


Asunto(s)
Pectinas , Proteómica , Pectinas/metabolismo , Proteómica/métodos , Especificidad por Sustrato , Poligalacturonasa/metabolismo , Poligalacturonasa/química , Beta vulgaris/química , Beta vulgaris/metabolismo , Aspergillus/enzimología
15.
Int J Biol Macromol ; 267(Pt 2): 131565, 2024 May.
Artículo en Inglés | MEDLINE | ID: mdl-38614184

RESUMEN

Endopolygalacturonases are crucial pectinases known for their efficient and sustainable pectin depolymerization activities. The present study identified a novel gene encoding endopolygalacturonase from an acidic mine tailing metagenome. The putative gene showed a maximum identity of 67.55 % with an uncharacterized peptide sequence from Flavobacterium fluvii. The gene was cloned and expressed in a heterologous host, E. coli. Biochemical characterization of the novel endopolygalacturonase enzyme variant (EPHM) showed maximum activity at 60 °C and at 5.0 pH, while retaining 50 % activity under the temperature and pH range of 20 °C to 70 °C for 6 h, and 3.0 to 10.0 for 3 h, respectively. The enzyme exhibited tolerance to different metal ions. EPHM was characterized for the depolymerization of methylated pectin into pectic oligosaccharides. Further, its utility was established for fruit juice clarification, as endorsed by high transmittance, significant viscosity reduction, and release of reducing sugars in the treated fruit juice samples.


Asunto(s)
Jugos de Frutas y Vegetales , Pectinas , Poligalacturonasa , Pectinas/metabolismo , Pectinas/química , Poligalacturonasa/metabolismo , Poligalacturonasa/química , Poligalacturonasa/genética , Jugos de Frutas y Vegetales/análisis , Concentración de Iones de Hidrógeno , Temperatura , Clonación Molecular , Polimerizacion , Oligosacáridos/química
16.
Int J Biol Macromol ; 267(Pt 1): 131469, 2024 May.
Artículo en Inglés | MEDLINE | ID: mdl-38604432

RESUMEN

Pectic polysaccharide is a bioactive ingredient in Chrysanthemum morifolium Ramat. 'Hangbaiju' (CMH), but the high proportion of HG domain limited its use as a prebiotic. In this study, hot water, cellulase-assisted, medium-temperature alkali, and deep eutectic solvent extraction strategies were firstly used to extract pectin from CMH (CMHP). CMHP obtained by cellulase-assisted extraction had high purity and strong ability to promote the proliferation of Bacteroides and mixed probiotics. However, 4 extraction strategies led to general high proportion of HG domain in CMHPs. To further enhance the dissolution and prebiotic potential of CMHP, pectinase was used alone and combined with cellulase. The key factor for the optimal extraction was enzymolysis by cellulase and pectinase in a mass ratio of 3:1 at 1 % (w/w) dosage. The optimal CMHP had high yield (15.15 %), high content of total sugar, and Bacteroides proliferative activity superior to inulin, which was probably due to the cooperation of complex enzyme on the destruction of cell wall and pectin structural modification for raised RG-I domain (80.30 %) with relatively high degree of branching and moderate HG domain. This study provided a green strategy for extraction of RG-I enriched prebiotic pectin from plants.


Asunto(s)
Bacteroides , Chrysanthemum , Pectinas , Pectinas/química , Chrysanthemum/química , Proliferación Celular/efectos de los fármacos , Celulasa/química , Celulasa/metabolismo , Solubilidad , Poligalacturonasa/química , Poligalacturonasa/metabolismo
17.
Food Funct ; 15(9): 4887-4893, 2024 May 07.
Artículo en Inglés | MEDLINE | ID: mdl-38597504

RESUMEN

Inhibition of galectin-3-mediated interactions by modified citrus pectin (MCP) could affect several rate-limiting steps in cancer metastasis, but the ability of MCP to antagonize galectin-8 function remains unknown. We hypothesized that MCP could bind to galectin-8 in addition to galectin-3. In this study, a combination of gradual ethanol precipitation and DEAE-Sepharose Fast Flow chromatography was used to isolate several fractions from MCP. The ability of these fractions to antagonize galectin-8 function was studied as well as the primary structure and initial structure-function relationship of the major active component MCP-30-3. The results showed that MCP-30-3 (168 kDa) was composed of Gal (13.8%), GalA (63.1%), GlcA (13.0%), and Glc (10.1%). MCP-30-3 could specifically bind to galectin-8, with an MIC value of 0.04 mg mL-1. After MCP-30-3 was hydrolyzed by ß-galactosidase or pectinase, its binding activity was significantly reduced. These results provide new insights into the interaction between MCP structure and galectin function, as well as the potential utility in the development of functional foods.


Asunto(s)
Citrus , Galectinas , Pectinas , Humanos , Proteínas Sanguíneas/química , Proteínas Sanguíneas/metabolismo , Citrus/química , Galectina 3/metabolismo , Galectinas/metabolismo , Galectinas/química , Pectinas/química , Pectinas/farmacología , Poligalacturonasa/química , Poligalacturonasa/metabolismo , Unión Proteica
18.
ACS Appl Mater Interfaces ; 16(12): 15143-15155, 2024 Mar 27.
Artículo en Inglés | MEDLINE | ID: mdl-38481099

RESUMEN

Realizing controllable input of botanical pesticides is conducive to improving pesticide utilization, reducing pesticide residues, and avoiding environmental pollution but is extremely challenging. Herein, we constructed a smart pesticide-controlled release platform (namely, SCRP) for enhanced treatment of tobacco black shank based on encapsulating honokiol (HON) with mesoporous hollow structured silica nanospheres covered with pectin and chitosan oligosaccharide (COS). The SCRP has a loading capacity of 12.64% for HON and could effectively protect HON from photolysis. Owing to the pH- and pectinase-sensitive property of the pectin, the SCRP could smartly release HON in response to a low pH or a rich pectinase environment in the black shank-affected area. Consequently, the SCRP effectively inhibits the infection of P. nicotianae on tobacco with a controlled rate for tobacco black shank of up to 87.50%, which is mainly due to the SCRP's capability in accumulating ROS, changing cell membrane permeability, and affecting energy metabolism. In addition, SCRP is biocompatible, and the COS layer enables SCRP to show a significant growth-promoting effect on tobacco. These results indicate that the development of a stimuli-responsive controlled pesticide release system for plant disease control is of great potential and value for practical agriculture production.


Asunto(s)
Plaguicidas , Plaguicidas/farmacología , Preparaciones de Acción Retardada/farmacología , Preparaciones de Acción Retardada/química , Poligalacturonasa , Agricultura , Pectinas
19.
Molecules ; 29(5)2024 Feb 28.
Artículo en Inglés | MEDLINE | ID: mdl-38474547

RESUMEN

Enzymatic hydrolysis using pectinase is critical for producing high-yield and quality sea buckthorn juice. This study determined the optimal temperature, time, and enzyme dosage combinations to guide manufacturers. A temperature of 60 °C, hydrolysis time of 3 h, and 0.3% enzyme dosage gave 64.1% juice yield-25% higher than without enzymes. Furthermore, monitoring physicochemical properties reveals enzyme impacts on composition. Higher dosages increase soluble solids up to 15% and soluble fiber content by 35% through cell wall breakdown. However, excessive amounts over 0.3% decrease yields. Pectin concentration also declines dose-dependently, falling by 91% at 0.4%, improving juice stability but needing modulation to retain viscosity. Electrochemical fingerprinting successfully differentiates process conditions, offering a rapid quality control tool. Its potential for commercial inline use during enzymatic treatment requires exploration. Overall, connecting optimized parameters to measured effects provides actionable insights for manufacturers to boost yields, determine enzyme impacts on nutrition/functionality, and introduce novel process analytical technology. Further investigations of health properties using these conditions could expand sea buckthorn juice functionality.


Asunto(s)
Hippophae , Poligalacturonasa , Poligalacturonasa/metabolismo , Hippophae/metabolismo , Temperatura , Frutas/química , Hidrólisis
20.
Food Chem ; 446: 138903, 2024 Jul 15.
Artículo en Inglés | MEDLINE | ID: mdl-38452507

RESUMEN

Cryoprotective effect and potential mechanism of soluble soybean polysaccharides (SSPS) and enzymatic hydrolysates on surimi was investigated. After hydrolysis, the molecular weight of SSPS significantly decreased, and the hydrolysates prepared by endo-polygalacturonase (EPG-SSPS) was the lowest (154 kDa). Infrared spectrum analysis revealed that enzymatic hydrolysis didn't alter the functional groups of SSPS, but it did augment the exposure to hydroxyl groups. Surimi containing 5 % EPG-SSPS had the lowest freezable water after 20 days of frozen storage. Furthermore, the 5 % EPG-SSPS group manifested the highest metrics in total sulfhydryl (8.0 × 10-5 mol/g), active sulfhydryl content (6.7 × 10-5 mol/g), Ca2+-ATPase activity, and exhibited the lowest level in carbonyl content, surface hydrophobicity (153 µg). Notably, the 5 % EPG-SSPS maintained the stability of protein structure. Conclusively, SSPS enzymatic hydrolysate using endo-polygalacturonase imparted superior cryoprotective effect on the myofibrillar protein of surimi, and the mechanism might be a decrease in molecular weight and exposure of hydroxyl groups.


Asunto(s)
Crioprotectores , Glycine max , Animales , Crioprotectores/química , Poligalacturonasa , Polisacáridos/farmacología , Polisacáridos/química , Congelación , Peces , Hidrolisados de Proteína/química
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