Cellulase-assisted Extraction Research Articles

Simultaneously efficient dissolution and structural modification of chrysanthemum pectin: Targeting at proliferation of Bacteroides

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.

Characterization and biological activities of polysaccharides extracted from Auricularia auricula with different extraction methods

Polysaccharides derived from Auricularia auricula exhibit diverse biological activities and hold significant potential for commercial utilization as functional food ingredients. In this investigation, polysaccharides from A. auricula were obtained using six extraction techniques (ammonium oxalate solution extraction, sodium hydroxide solution extraction, hot water extraction, pectinase and cellulase-assisted extraction, ultrasonic-assisted extraction, and microwave-assisted extraction). Subsequently, a comprehensive comparison was conducted to evaluate their physicochemical properties and biological functionalities. The ammonium oxalate solution extraction method yielded a higher extraction rate (11.76%) and polysaccharide content (84.12%), as well as a higher uronic acid content (10.13%). Although the six Auricularia polysaccharides had different molecular weight distributions, monosaccharide molar ratios, similar monosaccharide compositions, and characteristic functional groups of polysaccharides, they exhibited different surface morphology. In vitro assays showed that polysaccharides extracted by ammonium oxalate solution possessed good scavenging ability against DPPH free radical, hydroxyl free radical and superoxide anion free radical as well as reduction power of iron ion. At the same time, both polysaccharides extracted by ammonium oxalate solution and sodium hydroxide solution promoted NO production in mouse macrophages along with the secretion of cytokines TNF-α, IL-1β, and IL-6. These results indicated significant differences in the structure and characteristics among Auricularia polysaccharides prepared by various extraction methods, which may be related to the variety or origin of A. auricula; furthermore, their bioactivities varied accordingly in vitro assays where the ammonium oxalate solution extraction method was found more beneficial for obtaining high-quality bioactive Auricularia polysaccharides.

A polysaccharide from the seed of Gleditsia japonica var. delavayi: Extraction, purification, characterization and functional properties

Gleditsia japonica var. delavayi is widely distributed in Yunnan and Guizhou, China, and the perisperm of its seeds is frequently used as a traditional banquet sweet food with transparent color, smooth-tasting, low-fat, and high-carbohydrate. Herein, a cellulase-assisted extraction method was applied using response surface methodology to optimize the extraction of polysaccharides from the seeds of G. japonica var. delavayi. The highest extraction yield (82.23 ± 0.50%) was obtained under 5% cellulase, 176 mL/g liquid-to-material ratio, and enzymolysis parameters of 55 °C for 90 min at pH 4.5. The purified seed polysaccharide (PS) primarily contained galactomannan (weight average molecular weight, 42.74 kDa), which was linked by the monosaccharide residues 1,6-α-Galp, 1,4,6-α-D-Galp, 1,2,4,6-β-D-Manp, and 1,3,6-α-D-Galp. The PS obtained has a single-crystal structure, is thermodynamically stable, and its molecules aggregate in water. Furthermore, PS possesses high cholate binding capacity and a substantial inhibitory effect on α-glucosidase and α-amylase. Taken together, the findings of this report offer a theoretical foundation for developing and utilizing G. japonica var. delavayi.

Integration of phenylalanine precursor feeding and elicitor treatment in concert with the cellulase-assisted extraction approach to improve stilbene compound production

Integration of phenylalanine precursor feeding and elicitor treatment in concert with the cellulase-assisted extraction approach to improve stilbene compound production

Optimization of Cellulase-Assisted Extraction of Total Flavonoids from Equisetum via Response Surface Methodology Based on Antioxidant Activity

Flavonoids are one of the most essential compounds in various plants. This study used the cellulase-assisted method to extract flavonoid-enriched antioxidants from Equisetum arvense L. (EAL). According to a three-factor and three-level central composite design, the response surface methodology was used to obtain the maximum total flavonoid yield, thereby providing the most potent antioxidant activity of EAL extracts. The results indicated that the optimal condition for the total flavonoid extraction was 0.52% (base substrate) enzyme concentration and enzymolysis for 50.58 min at 49.03 °C. The extraction rate of flavonoids from horsetail reached 4.88 mg/g under these optimal conditions. The yield was 51.23% higher than that from the traditional solid–liquid extraction method. The highest DPPH-scavenging ability of the extracts obtained under 0.50% enzyme concentration and enzymolysis for 49.41 min at 46.59 °C was 77.36%. HPLC results revealed that the antioxidant substances had diverse flavonoids. Thus, the extraction condition was consistent for antioxidant activity and total flavonoids. Therefore, the current study provided an efficient method to extract flavonoid compounds from EAL, facilitating technical support for utilizing the plant.

Interaction between black mulberry pectin-rich fractions and cyanidin-3-O-glucoside under in vitro digestion

Pectic polysaccharides are potential candidates to enhance the stability of anthocyanins, and the interactions between pectin and anthocyanin are structure-dependent. In this study, mulberry pectic polysaccharides, extracted through different methods, and mulberry cyanidin-3- O -glucoside (C3G) were used to prepare complexes whose characterization and in vitro digestive properties were then investigated. The extracted mulberry polysaccharides were mostly made up of the monosaccharides galacturonic acid, glucose, mannose, galactose and rhamnose. The ratio of rhamnose to galacturonic acid (Rha/GalA) for the samples ranged from 0.06 to 0.56. The results showed that the main component of mulberry polysaccharide was pectin, and that the different preparation methods changed its chemical structure. Subsequently, different polysaccharide-C3G complexes were prepared: hot water-extracted polysaccharide-C3G (HWC), high temperature and pressure-extracted polysaccharide-C3G (HTPC), pectinase-extracted polysaccharide-C3G (PEC), cellulase-extracted polysaccharide (CEC)-C3G, pectin lyase-extracted polysaccharide-C3G (PLC) and compound enzyme-extracted polysaccharide-C3G (MC). Physicochemical analysis showed that the six pectic polysaccharide complexes bound to C3G to different degrees and in different ways. The binding affinities occur through ionic force, hydrogen bonding and hydrophobic interaction during digestion. Cellulase-assisted extraction greatly improved the binding capacity of obtained mulberry polysaccharides to C3G. High temperature and pressure-extracted mulberry polysaccharide, which was constituted of high proportion of galacturonic acid, greatly improved the bio-accessibility of C3G in small intestine. This study provides a theoretical basis from which to investigate the interaction mechanisms of anthocyanin and mulberry pectins during gastrointestinal digestion. The consequent influence of the interactions on fermentation patterns of the complex remains to be elucidated. • The mulberry pectins bind to cyanidin-3- O -glucoside in a structure-dependent manner. • The binding occurs via ionic force, H-bond and hydrophobic effect during digestion. • Stability of cyanidin-3- O -glucoside during digestion was enhanced by mulberry pectin. • Selected enzymatic treatments on mulberry may change the fate of its anthocyanins.

Enzyme-assisted extraction of anti-inflammatory compounds from habanero chili pepper (Capsicum chinense) seeds.

Capsaicinoids are the main bioactive compounds extracted from chili pepper seeds (CPSs) but other bioactive compounds such as phenolic compounds may be found. Enzyme-assisted extraction (EAE) improves the extraction of bioactive compounds from fruits and seeds. The aim of this study was to establish the cellulase-assisted extraction conditions of capsaicinoids and phenolic compounds from Habanero CPSs (Capsicum chinense) and to evaluate the anti-inflammatory activity of the obtained extracts on murine macrophages. EAE was performed using different temperatures (T1 = 30°C, T2 = 45°C and T3 = 60°C), enzyme concentrations (E1 = 2,500 UI/L and E2 = 250 UI/L), and extraction time periods (0-150 min). Total phenolic compounds were quantified using the Folin-Ciocalteu assay, capsaicin (CAP) and dihydrocapsaicin (DHC) contents were evaluated by HPLC, and anti-inflammatory activity was performed with Griess assay on murine macrophage RAW 264.7 cell culture. The highest phenolic compound content (337.96 mg GAE/L) was achieved at 30°C, 2,500 UI/L, and 150 min of extraction. The highest CAP content (310.23 μg/ml) was obtained at 45°C with 250 UI/L for 150 min, while for DHC (167.72 μg/ml), the conditions were 60°C, 2,500 UI/L, and 120 min. The highest anti-inflammatory response was obtained when 60°C, E2, and 150 min were used for the extraction, and nitric oxide (NO) production was reduced to 22.56%. Based on the results obtained in this research, EAE allowed the recovery of compounds with anti-inflammatory activity from CPS using water as a solvent. There was a correlation between the extraction of CAP and DHC. But although a moderate direct correlation between the concentration of capsaicinoids and total phenolic compounds (TPCs) and an inverse correlation of the presence of the bioactive compounds (TPC, CAP, and DHC) with the NO synthesis, these were not statistically significant. We demonstrated that Habanero seeds are an important raw material to recover anti-inflammatory compounds beyond capsaicinoids using water in EAE.

Structural characterization and antioxidant activity of polysaccharides extracted from Chinese yam by a cellulase-assisted method

Cellulase-assisted extraction of Chinese yam polysaccharides was investigated in the present study. The optimized method generated a polysaccharide yield of 10.6 % (w/w) from Chinese yam in 80 min using 8000 U/g of cellulose. Three polysaccharide fractions (CYP1, CYP2 and CYP3) were purified by Cellulose DEAE-52, and their structures and antioxidant activities were determined. There were five monosaccharides, namely arabinose, galactose, glucose, mannose, and galacturonic acid, in both CYP2 and CYP3, but in CYP1, there were only four monosaccharides, as galacturonic acid was missing. CYP1 and CPY2 were heteropolysaccharides; CYP3 was a homogeneous polysaccharide. Moreover, CYP3 showed the strongest antioxidant activity. Thus, methylation and NMR analysis were further applied to elucidate the detailed structure of CYP3. The backbone of CYP3 is composed of 1,2,4-linked-D- Glcp and 1,3,6-linked-D- Glcp. Bioactivity evaluation showed that CYP3 exhibited significant antioxidant activity in vitro, supporting the potential application of CYP3 as an antioxidant in food processing, pharmaceuticals and other applications.

Optimisation of cellulase-assisted extraction of laminarin from the brown seaweed Ecklonia maxima, using response surface methodology

Optimisation of cellulase-assisted extraction of laminarin from the brown seaweed Ecklonia maxima, using response surface methodology

Current Status of Mining, Modification, and Application of Cellulases in Bioactive Substance Extraction.

Cellulases have been used to extract bioactive ingredients from medical plants; however, the poor enzymatic properties of current cellulases significantly limit their application. Two strategies are expected to address this concern: (1) new cellulase gene mining strategies have been promoted, optimized, and integrated, thanks to the improvement of gene sequencing, genomic data, and algorithm optimization, and (2) known cellulases are being modified, thanks to the development of protein engineering, crystal structure data, and computing power. Here, we focus on mining strategies and provide a systemic overview of two approaches based on sequencing and function. Strategies based on protein structure modification, such as introducing disulfide bonds, proline, salt bridges, N-glycosylation modification, and truncation of loop structures, have already been summarized. This review discusses four aspects of cellulase-assisted extraction. Initially, cellulase alone was used to extract bioactive substances, and later, mixed enzyme systems were developed. Physical methods such as ultrasound, microwave, and high hydrostatic pressure have assisted in improving extraction efficiency. Cellulase changes the structure of biomolecules during the extraction process to convert them into effective ingredients with better activity and bioavailability. The combination of cellulase with other enzymes and physical technologies is a promising strategy for future extraction applications.

Optimization of Ultrasonic Cellulase-Assisted Extraction and Antioxidant Activity of Natural Polyphenols from Passion Fruit.

In this paper, ultrasonic cellulase extraction (UCE) was applied to extract polyphenols from passion fruit. The extraction conditions for total phenol content (TPC) and antioxidant activity were optimized using response surface methodology (RSM) coupled with a Box-Behnken design (BBD). The results showed that the liquid-to-solid ratio (X2) was the most significant single factor and had a positive effect on all responses. The ANOVA analysis indicated quadratic models fitted well as TPC with R2 = 0.903, DPPH scavenging activity with R2 = 0.979, and ABTS scavenging activity with R2 = 0.981. The optimal extraction parameters of passion fruit were as follows: pH value of 5 at 30 °C for extraction temperature, 50:1 (w/v) liquid-to-solid ratio with extraction time for 47 min, the experimental values were found matched with those predicted. Infrared spectroscopy suggested that the extract contained the structure of polyphenols. Furthermore, three main polyphenols were identified and quantified by HPLC. The results showed the content of phenolic compounds and antioxidant activity of the optimized UCE were 1.5~2 times higher than that determined by the single extraction method and the Soxhlet extraction method, which indicates UCE is a competitive and effective extraction technique for natural passion fruit polyphenols.

Optimization of Cellulase-assisted Extraction and Antioxidant Activity of Polysaccharides from Taraxacum Mongolicum Hand.-Mazz.

Polysaccharides from Taraxacum mongolicum were extracted using cellulase-assisted extraction technology. Based on the results of single factor experiments, three independent variables extraction time, temperature and pH were selected for investigation. Response surface methodology (RSM) was used to optimize the extraction conditions. The optimal conditions were as follows: extraction time, 105.07 min; extraction temperature, 47.49°C; extraction pH, 4.81. Under such conditions, the yield of polysaccharides was 13.75% (w/w), and the yield obtained by cellulase-assisted extraction method was remarkably higher than the hot water extraction. In vitro assays, scavenging activity of 1, 1-Diphenyl-2-picrylhydrazxyl (DPPH) radical and superoxide anion radical showed that the polysaccharides from T. mongolicum had definite antioxidant activities, and polysaccharides extracted from different parts of T. mongolicum had different antioxidant activity, which shows: inflorescence > leaf > root. These results exhibit that cellulase-assisted extraction is an advisable method for T. mongolicum polysaccharides and polysaccharides could be explored as potential antioxidant to use in medicine or functional food.

Optimization of aqueous enzyme‐assisted extraction of rosmarinic acid from rosemary ( Rosmarinus officinalis L.) leaves and the antioxidant activity of the extract

Rosemary (Rosmarinus officinalis L.) is a potential source of rosmarinic acid (RA). This study aimed to develop a novel aqueous enzymatic approach for extracting RA from rosemary leaves and evaluate the antioxidant activity of the RA-rich extract. The ability of several enzymes to release RA from plant tissues was investigated. Among the examined enzymes, cellulase A exhibited the highest extraction efficiency. Subsequently, aqueous cellulase A-assisted extraction was optimized. The optimal extraction conditions were as follows: an extraction time of 4.63 hr, a water-to-sample ratio of 28.69 ml/g, an enzyme concentration of 2.56%, and a temperature of 36.6°C, with the maximum RA content of 13.97 mg/g. The RA-rich extract exhibited potent antioxidant activity against the 2,2-diphenyl-1-picrylhydrazyl radical with an IC50 value of 532.01 μg/ml. Aqueous cellulase-assisted extraction is a green approach for RA extraction from rosemary leaves. Furthermore, the rosemary leaf extract is a promising antioxidant agent for future use. Practical applications RA is a potent antioxidant and is widely applied in the food and pharmaceutical industries. This study developed a new method involving the use of enzyme for ecofriendly extracting RA from rosemary leaves. The results indicated that the RA-rich extract is a potential antioxidant agent for future nutraceutical and pharmaceutical applications and the aqueous cellulase-assisted extraction is a practical and green process for RA extraction from rosemary leaves.

Cellulase-assisted extraction of phenolic compounds from rice bran (Oryza sativa L.): process optimization and characterization

This study investigated the improvement in the extraction of antioxidant phenolic compounds from a by-product of the food industry, rice bran, using hydrolytic enzymes. To this purpose, the optimization process of obtaining these compounds was carried out, as well as chemical characterization of the rice bran extracts by high performance liquid chromatography (HPLC) and the determination of their antioxidant activity. First, the parameters that affect the extraction process, such as the type on enzyme, temperature and pH of the solution, the extraction time and the enzyme concentration used for obtaining phenolic compounds were studied. The influence of these factors was later examined by characterizing the compounds obtained by HPLC, and the antioxidant activity of the extracts. The optimum conditions were: temperature 35 °C, pH 3.0 solution, extraction time 4 hours and cellulase content of 1.0%. In these conditions, extraction yield ranged from 3.45 ± 0.9 mg ferulic acid/100 g rice bran (without cellulase) to 14.3 ± 2.2 mg ferulic acid/100 g rice bran (with 1 % cellulase). Finally, the increases in antioxidant activity are lighter than those produced in the concentration of phenolic compounds, so we concluded that the enhancement in bioactivity is more limited when cellulase enzyme-assisted extractions were carried out in rice bran.

Study on extraction methods of polysaccharides from a processed product of Aconitum carmichaeli Debx.

Traditional Chinese medicine PaoTianXiong (PTX) is a processed product of Aconitum carmichaeli Debx. with polysaccharide as the main ingredient. The properties of PTX polysaccharide (PTXP) may be affected by different extraction methods. To develop and utilize PTXP better, it is of great significance to study the extraction methods of PTXP. Thus, we extracted PTXPs with dilute alkaline water extraction, ultrasound-assisted extraction, cellulase-assisted extraction, and hot water extraction (HWE), respectively. The characterizations of PTXPs extracted by different methods were analyzed based on purity determination, infrared analysis, molecular weight and monosaccharide composition. And antioxidant experiments of PTXPs were conducted. The results showed that PTXPs extracted by the four extraction methods were all glucan. After purification, the PTXPs showed similar antioxidant activity in vitro. The molecular weight of polysaccharides extracted by the cellulase-assisted method was different from that extracted by the other three methods. Our results showed that not only the yield but also the effect of extraction methods on the properties of PTXP should be considered when selecting the best extraction method. Therefore, HWE was considered to be the best extraction method of PTXP. The yield and purity of purified PTXP were 24.5% and 97.1%, respectively. The optimized extraction conditions were: an extraction temperature of 90 °C, extraction time of 2.17 h, solid–liquid ratio of 1 : 29 (g mL−1), and number of extractions of 2.

Flavonoids from the Zizania latifolia Bracts: Extraction Optimization, Antioxidant Activity and Exploration in Strawberry Preservation

Zizania latifolia bracts are Z. latifolia-based agricultural waste materials, distributed in various parts of the world. In the current study, the Response Surface Method (RSM) was used to optimize the cellulase-assisted extraction of Total Flavonoids from Z. latifolia bracts (TFZB). Then, the in vitro antioxidant capacity and the potential effect on strawberry preservation were assessed. The results showed that the optimal conditions for extracting TFZB was: Solid-to-liquid ratio of 1: 40 g/mL; ethanol concentration of 60% (v/v); cellulase dosage of 0.4% (w/w); incubation time of 2 h. Under these conditions, the yield of TFZB reached 0.49±0.01%. TFZB exerted certain antioxidant capacity. The free radical-scavenging rates against hydroxyl and DPPH radicals were 86±1.32% and 90±1.6% respectively at the concentration of 1.6 mg/mL and TFZB also exerted reducing power on Fe3+. Finally, the potential of TFZB on strawberry preservation was investigated from three aspects: Weightlessness rate, rotting rate and Vc content. It was found that the above-mentioned profiles were all improved after the strawberries sprayed with TFZB solution during the storage period. This study suggested that the TFZB may be developed as a novel antioxidant and fruits preservative, providing an evidence for the further utilization of Z. latifolia bracts.

Structure, physicochemical and bioactive properties of dietary fibers from Akebia trifoliata (Thunb.) Koidz. seeds using ultrasonication/shear emulsifying/microwave-assisted enzymatic extraction

Dietary fiber extracts from Akebia trifoliata (Thunb.) Koidz. seeds were generated using alkaline extraction, enzymatic hydrolysis as well as physical pretreatments including ultrasonication, shear emulsifying and microwave. Compared with the basic alkaline hydrolysis, cellulase-assisted extraction exhibited the highest water holding capacity (7.99 g/g), cation exchange capacity (0.55 mmol/g), glucose adsorption capacity (1352.33 μmol/g) and pancreatic lipase inhibition (37.69%). Ultrasonication pretreatment before both the preceding procedures increased the oil holding capacity (3.95 g/g) and α-amylase inhibition (6.81%) of the extracted fiber. Microwave pretreatment generated fiber possessing the greatest specific surface area (137.70 m2/kg), water swelling capacity (1.25 mL/g), adsorption capacity (4.14 mg/g, pH = 2) and sodium cholate adsorption capacity (38.68%). However, shear emulsifying gave the lowest crude yields (57.72%), glucose absorption capacity (22.09 mg/g), α-amylase inhibition (2.77%) and pancreatic lipase inhibition (22.61%) though it contained highest levels of soluble fiber (6.40 g/100 g) and lowest crystallinity (33.6%). A fuzzy statistical evaluation indicated that ultrasonication assisted enzymatic hydrolysis was the most efficient improvement in the overall properties of the dietary fiber. These results provide valuable information for the potential use of A. trifoliata seeds as a new food ingredient and for extraction of dietary fiber from this plant.

Chemical composition and cellulase-assisted extraction of total saponins from Gomphrena celosioides Mart.

Chemical composition and cellulase-assisted extraction of total saponins from Gomphrena celosioides Mart.

Effects of different extraction techniques on physicochemical properties and biological activities of polysaccharides from Fritillaria pallidiflora Schrenk

Abstract In this study, polysaccharides were extracted using different techniques including ultrasound-assisted extraction (UA), cellulase-assisted extraction (CA), cellulase-ultrasound-assisted extraction (CUA) and pectinase-ultrasound-assisted extraction (PUA) from Fritillaria pallidiflora Schrenk. Extraction yields, chemical composition, physicochemical, structural properties and antioxidant activities of the polysaccharides were compared. Results shows that polysaccharides extracted with CUA method has the highest extraction yield (20.65 ± 0.78%), including carbohydrate (56.16 ± 0.38%) and uronic acid (28.34 ± 0.23%), demonstrates better solubility and oil holding capacity as compared with others. The polysaccharides composed of rhamnose, lyxose, arabinose, xylose, mannose, glucose and galactose with different molar ratio. Fourier transform infrared spectroscopy showed key functional groups whereas scanning electron microscopy, X-ray diffraction and Congo red test of the extracts obtained with different techniques showed characteristic structures of polysaccharides. Biological activity tests showed that polysaccharides extracted with UA method exhibited strong scavenging activity against ABTS and hydroxyl free radicals. The results indicated that different extraction techniques had certain effects on chemical composition, physicochemical properties and bioactivities of polysaccharides.

Optimization for the Extraction of Polysaccharide from Walnut (Juglans regia L.) Leaves: Antioxidant Activities in Vitro

Cellulase-assisted extraction of Walnut Leaf Polysaccharide (WLP) was studied. Single factor test and response surface design were employed to optimize the technological conditions. The consequences indicated that the highest WLP yield of 5.21% was achieved with raw material ratio of 50 mL/g, cellulase dose of 41488.52 U/g, enzymolysis pH of 5.88, enzymolysis time of 29.74 min and enzymolysis temperature of 52.07°C. Moreover, in vitro antioxidant assays revealed that WLP has significant eliminating capability against 1, 1-diphenyl-2-picrylhydrazyl (DPPH), 2, 2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and hydroxyl free radicals in a dose-dependent manner and exhibited a strong reducing power. These results suggest that WLP could be developed as a promising natural antioxidant agent in the pharmaceutical and functional food industries.