Enzymatic Assisted Extraction Research Articles

Differences in physicochemical properties of pectin extracted from pomelo peel with different extraction techniques

In order to obtain high yield pomelo peel pectin with better physicochemical properties, four pectin extraction methods, including hot acid extraction (HAE), microwave-assisted extraction (MAE), ultrasound-assisted extraction, and enzymatic assisted extraction (EAE) were compared. MAE led to the highest pectin yield (20.43%), and the lowest pectin recovery was found for EAE (11.94%). The physicochemical properties of pomelo peel pectin obtained by different methods were also significantly different. Pectin samples obtained by MAE had the highest methoxyl content (8.35%), galacturonic acid content (71.36%), and showed a higher apparent viscosity, thermal and emulsion stability. The pectin extracted by EAE showed the highest total phenolic content (12.86%) and lowest particle size (843.69 nm), showing higher DPPH and ABTS scavenging activities than other extract methods. The pectin extracted by HAE had the highest particle size (966.12 nm) and degree of esterification (55.67%). However, Fourier-transform infrared spectroscopy showed that no significant difference occurred among the different methods in the chemical structure of the extracted pectin. This study provides a theoretical basis for the industrial production of pomelo peel pectin.

Brewers' spent grain proteins: The extraction method determines the functional properties

Brewers' spent grains (BSG) are a major byproduct from the brewing industry that are currently discarded or used as animal feed. This study systematically explored the effects of BSG protein extraction methods using alkali, ethanol or enzymes on the protein's structure, composition and function. Proteins extracted at pH 12 were partially unfolded and glutelin-rich while those extracted by 55% ethanol containing 2-mercaptoethanol were highly aggregated and hordein-rich. Enzymatic-assisted extraction resulted in peptides below 10 kDa that had no distinct structural elements. Being completely water-soluble, these peptides gave good emulsifying properties (activity: 83 m2/g protein; stability: 35 min) and a high antioxidant activity. On the other hand, alkali-extracted and ethanol-extracted proteins were mostly insoluble but exhibited high water holding capacities (2.5–4.0 g/g) that enabled gelation. The results highlighted that selection of the extraction method is critical, as the composition, structure and function of proteins are modified, which affects its potential applications. Industrial relevanceAs a major side stream from the production of beer, brewers' spent grains are present in huge amounts. These residues are a promising source of alternative proteins, but before separating them, it is important to first consider an extraction method that does not compromise on its functionality. This study offered insights into the influence of extraction methods on the structure and function of the resulting protein concentrates as well as its future applications.

Extracting bioactive compounds and proteins from Bacopa monnieri using natural deep eutectic solvents.

This study employed novel extraction methods with natural deep eutectic solvents (NADES) to extract bioactive compounds and proteins from Bacopa monnieri leaves. The conditional influence of ultrasonic-assisted extraction (UAE), microwave-assisted extraction (MAE), and enzymatic-assisted extraction (EAE) on the recovery efficiency of phenolics, proteins, flavonoids, and terpenoids was evaluated. The conditions of UAE were 50 mL/g LSR, 600W of ultrasonic power, and 30% water content with 40°C for 1 min to obtain the highest bioactive compounds and protein contents. The conditions of MAE were 40 mL/g LSR, 400W of microwave power with 30% water content for 3 min to reach the highest contents of biological compounds. The conditions of EAE were 30 mL/g of LSR, 20 U/g of enzyme concentration with L-Gly-Na molar ratio at 2:4:1, and 40% water content for 60 min to acquire the highest bioactive compound contents. Scanning electron microscopy (SEM) is employed to analyze the surface of Bacopa monnieri leaves before and after extraction. Comparing seven extraction methods was conducted to find the most favorable ones. The result showed that the UMEAE method was the most effective way to exploit the compounds. The study suggested that UMEAE effectively extracts phenolics, flavonoids, terpenoids, and protein from DBMP.

Determination of process parameters and precipitation methods for potential large-scale production of sugar beet leaf protein concentrate.

Sugar beet is one of the most produced industrial plants in the world, and during manufacturing it produces a large quantity of leaf waste. Because this waste is rich in protein, this study aimed to identify an efficient method for producing large-scale protein concentrate from sugar beet leaves. Results showed that protein extraction from fresh leaves was more effective than from dried leaves. Maximum protein extraction was achieved at pH 9, compared with pH 7 or 8. Blanching as a pretreatment reduced protein yield during isoelectric precipitation, with a yield of 2.31% compared to 20.20% without blanching. Consequently, blanching was excluded from the extraction process. After extraction, isoelectric precipitation, heat coagulation, and isoelectric-ammonium sulfate precipitation were compared. Although the latter resulted in the highest protein yield, Fourier transform infrared analysis revealed that excessive salt was not removed during dialysis, making it unsuitable for scale-up due to its additional cost and complexity. Therefore, isoelectric precipitation was selected as the appropriate method for protein precipitation from sugar beet leaves. To increase yield, extractions were assisted by ultrasound or enzyme addition. Ultrasound-assisted extraction resulted in an increased protein yield from 20.20% to 28.60%, while Pectinex Ultra SP-L-assisted extraction was the most effective, increasing protein yield from 20.20% to 38.09%. Proteins were extracted from fresh sugar beet leaves using optimum conditions (50 °C, 30 min, pH 9) and precipitated at isoelectric point, with enzymatic-assisted extraction yielding the maximum protein recovery. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Enhancing Efficiency of Enzymatic-Assisted Extraction Method for Evaluating Bioactive Compound Analysis in Mulberry: An Optimization Approach

The present investigation endeavors to optimize a method based on enzyme-assisted extraction for the efficient retrieval of bioactive compounds from mulberry, leveraging its notable health-promoting properties. A combined approach of Plackett–Burman design followed by Box–Behnken design was employed for determining the crucial extraction parameters and subsequently, refining the process. Optimal conditions consisted of heating 0.15 g of mulberry at 40 °C, using 15 mL of 70% EtOH as a solvent at pH 4, 38.46 enzyme units per g of sample, and shaking at 200 rpm. The optimum extraction time study revealed that 5 min of extraction was sufficient to reach the maximum concentration of the bioactive compound. The repeatability and intermediate precision assessment exhibited a coefficient of variation below 5%. Among the diverse mulberry varieties scrutinized, Morus nigra showed the highest anthocyanin content (27.90 ± 2.14 mg/100 g), while Morus rubra showed the highest concentration of phenolic compounds (121.10 ± 19.56 mg/100 g). Moreover, the extracted compounds showcased significant antioxidant and antimicrobial properties.

Upcycling mango peels into a functional ingredient by combining fermentation and enzymatic-assisted extraction

This study aims at upcycling mango peels by a sequential application of enzymatic hydrolysis, using Viscozyme and Pectinex at 50 °C for 2 h; and fermentation, using L. plantarum and B. animalis at 48 h for 37 °C. The use of Viscozyme led to a considerable increase in the concentration of galacturonic and glucuronic acids in the unfermented samples (308.96 and 12.97 mg/100 ml higher than control, respectively), whereas the use of Pectinex resulted in higher oligosaccharide solubilization (5.3 % more than control). None of the enzymes influenced microbiological growth. The recovery of gallic acid aglycone increased 17-fold over the control when Pectinex and B. animalis were used. Similarly, the recovery of mangiferin aglycone increased by 60 % after fermentation by either bacteria. The results indicate that this sequential processing strategy might be utilized to extract phenolic aglycones and produce functional ingredients from mango peels.

Effects of different extraction techniques on the structural, physicochemical, and bioactivity properties of heteropolysaccharides from Platycodon grandiflorum roots

In this study, two different extraction methods, enzymatic-assisted extraction (EAE) and ultrasonic-assisted extraction (UAE), were used to extraction of polysaccharides from the roots of Platycodon grandiflorum (PGR), and the preliminary characterization of the purified polysaccharide (PGP1 extracted with EAE and PGP2 extracted with UAE) were analyzed. The data implied that the yield of the polysaccharide obtained by UAE (12.01 ± 0.11%) was significantly higher than that of EAE (9.06 ± 0.07%). Meanwhile, Chemical composition analysis revealed that both PGP1 (6.6 kDa) and PGP2 (5.4 kDa) were comprised of eight monosaccharides with different molar ratios. Preliminary characterization analysis including FT-IR spectra, NMR analysis, X-ray diffraction, Congo red experiment, and thermogravimetric analysis were investigated. The result illustrated that PGP1 and PGP2 have similar chemical composition and functional groups, and both have crystalline and triple helix structure. Moreover, PGP2 demonstrated better water (4.94 ± 0.14 g/g) and oil (2.69 ± 0.016 g/g) retention capacity than PGP1, and also showed higher bile binding ability and pancreatic lipase inhibition activity. However, PGP1 presented higher inhibitory activities against α-amylase and α-glucosidase. Based on the above, these findings indicated that the two novel polysaccharides are worthy being developed as natural hypoglycemic and hypolipidemic agents.

Protease-Assisted Mild Extraction of Soluble Fibre and Protein from Fruit By-Products: A Biorefinery Perspective.

By-products from the fruit supply chain, especially seeds/kernels, have shown great potential to be valorised, due to their high content of macronutrients, such as lipids, protein, and fibre. A mild enzymatic assisted extraction (EAE) involving the use of a protease was tested to evaluate the feasibility of a cascade approach to fractionate the main fruit by-products components. Protease from Bacillus licheniformis (the enzyme used in the AOAC 991.43 official method for dietary fibre quantification) was used, and besides protein, the conditions of hydrolysis (60 °C, neutral pH, overnight) allowed us to dissolve a portion of soluble fibres, which was then separated from the solubilized peptide fraction through ethanol precipitation. Good protein extraction yields, in the range 35-93%, were obtained. The soluble fibre extraction yield ranged from 1.6% to 71% depending on the by-product, suggesting its applicability only for certain substrates, and it was found to be negatively correlated with the molecular weight of the fibre. The monosaccharide composition of the soluble fibres extracted was also diverse. Galacturonic acid was present in a low amount, indicating that pectin was not efficiently extracted. However, a predominance of arabinose and galactose monomers was detected in many fractions, indicating the isolation of a fruit soluble fibre portion with potential similarity with arabinogalactans and gum arabic, opening up perspectives for technological applications. The residual solid pellet obtained after protease assisted extraction was found to be an excellent fibre-rich substrate, suitable for being subjected to more "hard" processing (e.g., sequential pectin and hemicellulose extraction) with the objective to derive other fractions with potential great added economic value.

Composition of Nonextractable Polyphenols from Sweet Cherry Pomace Determined by DART-Orbitrap-HRMS and Their In Vitro and In Vivo Potential Antioxidant, Antiaging, and Neuroprotective Activities

Sweet cherry pomace is an important source of phenolic compounds with beneficial health properties. As after the extraction of phenolic compounds, a phenolic fraction called nonextractable polyphenols (NEPs) remains usually retained in the extraction residue, alkaline and acid hydrolyses and enzymatic-assisted extraction (EAE) were carried out in this work to recover NEPs from the residue of conventional extraction from sweet cherry pomace. In vitro and in vivo evaluation of the antioxidant, antihypertensive, antiaging, and neuroprotective capacities employing Caenorhabditis elegans was achieved for the first time. Extractable phenolic compounds and NEPs were separated and identified by families by high-performance thin-layer chromatography (HPTLC) with UV/Vis detection. A total of 39 phenolic compounds were tentatively identified in all extracts by direct analysis in real-time high-resolution mass spectrometry (DART-Orbitrap-HRMS). EAE extracts presented the highest in vitro and in vivo antioxidant capacity as well as the highest in vivo antiaging and neuroprotective capacities. These results showed that NEPs with interesting biological properties are retained in the extraction residue, being usually underestimated and discarded.

In vitro assessment of the bioavailability of bioactive non-extractable polyphenols obtained by pressurized liquid extraction combined with enzymatic-assisted extraction from sweet cherry (Prunus avium L.) pomace

In vitro digestion and absorption simulation processes of non-extractable polyphenols (NEPs) obtained by pressurized liquid extraction combined with enzymatic-assisted extraction with Promod enzyme (PLE-EAE) from the residue of conventional extraction of sweet cherry pomace were studied. In general, total phenolic and proanthocyanidin contents decreased in each phase of the digestion. However, the antioxidant capacity increased when the digestion process progressed. In addition, the highest total phenolic and proanthocyanidin contents and antioxidant capacity were obtained in the absorbed fraction. NEPs from PLE-EAE extract, digestive fractions, absorbed and unabsorbed fractions were analyzed by ultra-high-performance liquid chromatography coupled to electrospray ionization quadrupole Exactive-Orbitrap mass spectrometry (UHPLC-ESI-Q-Orbitrap-MS). Fifteen NEPs were identified in the intestinal fraction and five in the absorbed fraction after the digestion process. Results obtained in this study define for the first time the bioavailability of antioxidant NEPs obtained from sweet cherry pomace.

Effect of Extraction Procedures with Ultrasound and Cellulolytic Enzymes on the Structural and Functional Properties of Citrus grandis Osbeck Seed Mucilage.

The structural and functional properties of Citrus grandis Osbeck (CGO) seed mucilage by different extraction practices, including conventional citrate buffer, ultrasonic-assisted (UAE), enzymatic-assisted extraction (EAE) with cellulase or Celluclast® 1.5 L and various ultrasonic-assisted enzymatic extraction (UAEE) procedures were investigated. It was found that CGO seed from agricultural and processing byproducts is an excellent new source of high methoxyl pectin with quite high intrinsic viscosity (about 108.64 dL/g) and molecular weight (about 1.9 × 106) as compared with other pectin sources. UAEE with Celluclast® 1.5 L enhanced the extraction yield most pronouncedly (about 2.3 times). Moreover, the monosaccharide composition of CGO seed mucilage is least affected by EAE with Celluclast® 1.5 L. In contrast, EAE with cellulase dramatically reduces the galacturonic acid (GalA) content to less than 60 molar%, and increases the glucose (Glc) content pronouncedly (to about 40 molar%), which may be considered as an adverse effect in terms of pectin purity. Though extraction procedures involved with ultrasound and cellulolytic enzymes generally show a decrease in GalA contents, weight average molar mass and intrinsic viscosity, EAE with Celluclast® 1.5 L is least affected, followed by UAE and UAEE with Celluclast® 1.5 L. These features can be leveraged in favor of diversified applications.

Rapid fingerprinting of extractable and non-extractable polyphenols from tropical fruit peels using direct analysis in real time coupled to orbitrap mass spectrometry

A simple and rapid direct analysis in real-time coupled to high-resolution mass spectrometry (DART-HRMS) methodology was developed to generate the extractable and non-extractable polyphenols (NEPs) fingerprint for four different passion fruits, G. mangostana, and A. squamosa peels as case-study to investigate the influence of alkaline hydrolysis and enzymatic-assisted extraction (EAE) on the recovery of NEPs. The extraction residue obtained after these treatments was also analyzed by DART-HRMS. Data compiled from DART-HRMS mass spectra were processed with principal component analysis to discriminate among the different treatments. EAE with Depol enzyme enabled to obtain NEPs with the highest signal intensity in DART-HRMS analysis from all peels except for P. edulis and A. squamosa peels. In these two cases, NEPs were better extracted by EAE with Promod enzyme and alkaline hydrolysis. Results showed that the applied treatments were efficient to extract NEPs since their signal intensities in the extraction residues were very low compared with their extracts.

Pressurized Liquid Extraction Combined with Enzymatic-Assisted Extraction to Obtain Bioactive Non-Extractable Polyphenols from Sweet Cherry (Prunus avium L.) Pomace.

Sweet cherry generates large amounts of by-products within which pomace can be a source of bioactive phenolic compounds. Commonly, phenolic compounds have been obtained by conventional extraction methodologies. However, a significant fraction, called non-extractable polyphenols (NEPs), stays held in the conventional extraction residues. Therefore, in the present work, the release of NEPs from cherry pomace using pressurized liquid extraction (PLE) combined with enzyme-assisted extraction (EAE) using PromodTM enzyme is investigated for the first time. In order to study the influence of temperature, time, and pH on the NEPs extraction, a response surface methodology was carried out. PLE-EAE extracts displayed higher TPC (75 ± 8 mg GAE/100 g sample) as well as, PA content, and antioxidant capacity than the extracts obtained by PLE (with a TPC value of 14 ± 1 mg GAE/100 g sample) under the same extraction conditions, and those obtained by conventional methods (TPC of 8.30 ± 0.05 mg GAE/100 g sample). Thus, PLE-EAE treatment was more selective and sustainable to release NEPs from sweet cherry pomace compared with PLE without EAE treatment. Besides, size-exclusion chromatography profiles showed that PLE-EAE allowed obtaining NEPs with higher molecular weight (>8000 Da) than PLE alone.

Optimization of extractions of eumelanin from cuttlefish ink and the hypoglycemic effects: In vitro enzyme inhibitory activity and glucose consumption in HepG2 cells

Pigment compounds with hypoglycemic effects are a widely researched topic, but there is almost no research evaluation on eumelanin in ink. Melanin in cuttlefish juice is used as an antibacterial, hemostatic, and other active natural medicine and food additive. This study is the first to compare the extraction method of alkali-extracted eumelanin and enzyme-extracted eumelanin and to analyze their hypoglycemic activities. The structures of the extracted samples were analyzed by Fourier Transform infrared spectroscopy, Ultraviolet-visible spectroscopy (UV-vis), X-ray, and scanning electron microscope (SEM). The purity of the extracted samples was determined by high-performance liquid chromatography (HPLC). The results showed that the enzyme extraction yield and purity of eumelanin were 88.3% and 91.62%, and the alkali extraction yield and purity of eumelanin were 16.82% and 58.57%, respectively. Enzyme extraction of eumelanin can significantly inhibit α-glycosidase enzyme and α-amylase activity, promote the utilization of glucose in cells, and increase hepatic glycogen synthesis. Eumelanin (0.8 mg/ml) was selected as the effective concentration. This study provides a technical reference for the extraction, determination, and hypoglycemic activity of eumelanin. Novelty impact statement This study is the first to compare the extraction method of alkali-assisted extraction of eumelanin and enzymatic-assisted extraction of eumelanin and to analyze their hypoglycemic activities. The extraction yield and purity of enzyme-assisted extraction of eumelanin extraction is much higher than that of alkali-assisted extraction of eumelanin. Eumelanin from squid ink has the effect of accelerating the utilization of glucose and the effect of enzyme-assisted extraction of eumelanin is slightly better than alkali-assisted extraction of eumelanin.

Identification of novel antihypertensive peptides from wine lees hydrolysate

Enzymatic-assisted extraction using Flavourzyme® has been demonstrated to be a useful methodology to obtain wine lees (WL) enriched in phenolic compounds and with enhanced antihypertensive activity. Nevertheless, taking into account that Flavourzyme® possess proteolytic activity, the release of bioactive peptides should not be ruled out. In this study, we investigate the presence of antihypertensive peptides in the WL hydrolysate. Peptides were separated into fractions by ultrafiltration and RP‐HPLC. Next, peptide identification by nano-HPLC-(Orbitrap)MS/MS was performed in the fractions showing the highest angiotensin-converting enzyme inhibitory (ACEi) activities. Six peptides were identified; three of them showing ACEi (IC50) values lower than 20 µM. The peptide antihypertensive effect was evaluated in spontaneously hypertensive rats at an oral dose of 10 mg/kg bw. Peptides FKTTDQQTRTTVA, NPKLVTIV, TVTNPARIA, LDSPSEGRAPG and LDSPSEGRAPGAD exhibited antihypertensive activity, confirming that they could contribute to the blood pressure-lowering effect of the WL hydrolysate. These peptides have a great potential as functional ingredients to manage hypertension.

Targeting the Nutritional Value of Proteins From Legumes By-Products Through Mild Extraction Technologies

Legumes have been known for centuries for their good nutritional properties. Unfortunately, during processing, from 5 to 25% of this production is wasted, generating by-products that can still be a rich source of useful compounds, such as proteins, which can still be used in food and feed formulations. The choice of the extraction technique is important to preserve the nutritional value of proteins since drastic conditions of pH and/or temperature could damage them. In this work, two mild extraction techniques (direct assisted extraction—DAE and enzymatic assisted extraction—EAE) were applied for protein extraction from legume by-products obtained from agro-industrial processes. The quality of proteins was evaluated considering protein integrity [SDS-PAGE, degree of hydrolysis (DH), free amino acid content, racemization degree] and nutritional features [amino acid score (AAS), digestibility]. Direct assisted extraction is the technique that has best preserved protein integrity (1–5% DH and free amino acid content <1%), The digestibility of proteins extracted with EAE is higher (no protein bands detected in SDS-PAGE) than with the one of DAE extracts, making this technique particularly suitable for those food and feed formulation were a high digestibility of proteins is required.

Novel food colorants from tomatoes: Stability of carotenoid-containing chromoplasts under different storage conditions

Despite years of research, the application of natural pigments as food ingredients is still limited by their low stability. This research investigated the storability of carotenoid-containing chromoplasts, as novel natural food colorant from unsold tomatoes.The 30-day stability of chromoplasts (Chr), in comparison to free carotenoid-based extract (Free car), was investigated by the kinetic degradation model of carotenoids (UV–visible spectrophotometer assay), and by the color attributes as CIELab* parameters [i.e. total color difference (ΔE); red-yellow ratio (a*/b*) and chroma value (C*)], under dark and UV-light exposure at different temperatures (4, 25 and 40 °C).Regardless of the different storage conditions (temperature and UV-light exposure), the lowest pigment degradation rates were found for Chr [0.014 days−1 (25 °C in the dark) - 0.057 days−1 (40 °C under UV-light)] rather than Free car [0.018 days−1 (25 °C in the dark) - 0.071 days−1 (40 °C under UV-light)]. Furthermore, Chr better retained the colorimetric parameters, especially at 4 °C (ΔE ~ 4.0) and 25 °C (ΔE ~ 4.5) in the dark. These results proving that a tailored enzymatic-assisted extraction allowed the recovery of stable natural red-orange pigments better preserved by their chromoplast envelop.

Life Cycle Assessment of Chemical vs Enzymatic-Assisted Extraction of Proteins from Black Soldier Fly Prepupae for the Preparation of Biomaterials for Potential Agricultural Use

The cradle-to-grave life cycle assessment (LCA) was applied to the preparation of biomaterials derived from proteins, extracted from black soldier fly (BSF) prepupae, after the larvae were reared o...

Enzyme-assisted extraction of carotenoids and phenolic compounds from sunflower wastes using green solvents.

The aim of this work is to develop an optimized enzymatic assisted extraction methodology to extract carotenoids and phenolic compounds from sunflower wastes (petals and florets) using natural hydrophobic green solvents. Several natural green hydrophobic solvents were used as well as natural hydrophobic eutectic solvents composed of d,l-menthol and different acids, with different hydrophobicity. The multi-enzyme complex Viscozyme® was used to disrupt the cell wall of petals and disc florets. The extracted carotenoids content into the hydrophobic phase was quantified using UV-Vis spectrophotometry and the carotenoids profile was studied using high-performance liquid and thin layer chromatography. The amount of total sugars in the aqueous phase was also analyzed using the dinitrosalicylic acid (DNS) method to infer about the enzymatic action in cell wall. Phenolic compounds also in the aqueous phase were analyzed by Folin Denis method. The eutectic solvent d,l-menthol:d,l-lactic acid (M:HLac) (1:2) was the best solvent for extraction of carotenoids from sunflower wastes, with 147ppm of carotenoids extracted, in comparison to 115ppm obtained with the standard solvent, n-hexane. In what concerns phenolic compounds, M:HLac was again better than the standard solvent. The use of the multi-enzyme complex Viscozyme® had different responses, depending on the solvent tested. For the green solvent M:HLac, the enzyme improved the carotenoids extraction, achieving 335ppm of carotenoids in the extract. The role of enzyme, solvent, water and sunflower quantity in the carotenoid extraction was evaluated and optimized through a central composite rotatable design (CCRD), using the M:HLac as solvent. According to the analysis of CCRD, the most efficient extractions were carried out using more solvent and less raw material, whose best result reached 1449 mg carotenoids/100 g biomass ppm of carotenoids. This work emphasizes the possibility of developing more sustainable enzyme-assisted separation processes, through the substitution of toxic solvents with natural, environmentally friendly, solvents.

Process optimization for enzymatic assisted extraction of anthocyanins from the mulberry wine residue

In order to improve the utilization value of the by-product of mulberry, the remaining residue of mulberry wine was used as raw materials to extract anthocyanins from the residue of mulberry wine by enzymatic method, and the response surface method was used to optimize the extraction process. Firstly, the single factor experiment was carried out by controlling four factors: liquid-solid ratio, extraction time, temperature and pH value. Then, Box-Behnken was used to design a three-factor, three-level experiment. Finally, the response surface method was used to optimize the extraction process. The results showed that the optimal extraction process of pectinase was the liquid-solid ratio of 1:20, the extraction time was 58 min, the pH was 5.90 and the extraction temperature was 45 °C, and the extraction effect was the best. The highest anthocyanin content in the extract was 6.040 mg/g.