Antioxidant activity of partially characterized polysaccharides from the edible mushroom Pleurotus djamor var. roseus

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J. Mushrooms 2021 September, 19(3):140-149 http://dx.doi.org/10.14480/JM.2021.19.3.140 Print ISSN 1738-0294, Online ISSN 2288-8853

© The Korean Society of Mushroom Science

Jegadeesh Raman(Post-Doc), Archana Sivakumar(Student), Hariprasath Lakshmanan(Professor), Nanjian Raaman(Professor), and Hyun-Jae Shin(Professor)

*Corresponding author E-mail : [email protected] Tel : +82–62-230-7518 Received August 2, 2021 Revised August 25, 2021 Accepted September 1, 2021

Antioxidant activity of partially characterized polysaccharides from the edible mushroom Pleurotus djamor var. roseus

Jegadeesh Raman^1,2, Archana Sivakumar², Hariprasath Lakshmanan², Nanjian Raaman², and Hyun-Jae Shin*

1Department of Chemical Engineering, Graduate School of Chosun University, 309 Pilmun-daero, Dong-gu, Gwangju 61452, Republic of Korea

2Centre for Advanced Studies in Botany, University of Madras, Guindy Campus, Chennai 600 025,Tamil Nadu, India

3Department of Biochemistry, Karpagam Academy of Higher Education, Coimbatore 641021, India

ABSTRACT: Mushroom-derived polysaccharides, which are the primary bioactive constituents, are beneficial for human health.

Polysaccharides have immuno-modulation, antitumor, and antioxidant properties. Additionally, they have antiviral properties and protect against chronic radiation stress. In this study, high yield water-soluble polysaccharides were obtained from Pleurotus djamor var. roseus basidiocarps. The crude polysaccharide (CP) was extracted sequentially by hot water and ethanol precipitation.

The yield of the brown CPs was 5.6% dw. Diethylaminoethyl cellulose and Sepharose-6B column chromatography of CPs generated several fractions. Total glucan content was determined in all the fractions. The F1 fraction displayed the highest sugar content and was considered as a purified polysaccharide (PP). The total glucan and β-glucan content in the four fractions ranged between 76.85-2.95% and 75.08-1.46%, respectively. The yield of the PPs was 300 mg, and it was obtained as a white powder.

The PPs were characterized by Fourier-transform infrared spectroscopy (FTIR) and thin-layer chromatography. The FTIR spectral details confirmed the presence of a xylopentose polysaccharide. The antioxidant activity of the PPs was evaluated using in vitro 1,1-diphenyl-2-picryl-hydrazyl (DPPH) free radical scavenging assay and superoxide radical scavenging assay. The PPs showed strong DPPH free radical and superoxide anion radical scavenging activities in a dose-dependent manner. Purified PPs free of phenolics, protein, and carbohydrates were mainly responsible for the radical scavenging activity. The data suggest the potential of PPs as natural antioxidants.

KEYWORDS: Mushroom, Pink oyster, Pleurotus, Polysaccharide, Antioxidant

INTRODUCTION

Many wild and cultivated mushroom species have been used as food and medicine for millennia. They can produce many bioactive molecules, such as polysaccharides, terpenoids, proteins, protein-bounded polysaccharides,

and steroids (Camelini et al., 2014; Arteiro et al., 2012;

Jegadeesh et al., 2020). Mushroom-derived polysaccharides are considered nontoxic and showed tremendous biological and functional properties (Meng et al., 2016). The polysaccharide is an active constituent in the fungal cell walls and is mainly made with β-D-glucose monomers. In recent, mushroom polysaccharides have much attention to biological application, and even many commercial species are explored (Seedevi et al., 2019). Friedman (2015) reported that high molecular weight β-glucan exhibits a broad spectrum of biological activity. However, the natural substance of cell wall polysaccharides isolated from the mushrooms showed potential antioxidant and antitumor- related activity.

The antioxidant potential observed in the mushroom is part of their natural defense mechanism against noxious events causing oxidative damage. Free radicals are generated during the oxidative metabolites process in living cells. They are unstable and create oxidative stress, which may have stimulated various diseases like cancer, diabetes, atherosclerosis, neurodegeneration, and aging (Ray et al.,

2012). Antioxidant-rich food and supplements could prevent/neutralize oxidative stress and even many chronic diseases. Thus, the requirement for natural and alternative sources of antioxidant-rich food is in demand.

In recent years, edible and medicinal mushrooms have attracted commercial sources of antioxidants (Khatua et al., 2013; Ren et al., 2015; Fan et al., 2012). Among the edible mushrooms, Pleurotus species are prevalent for their culinary and medicinal properties. Moreover, they are widely cultivated and common edible mushrooms, which are cultivated in largescale worldwide. They are mainly consumed due to their excellent nutritive and medicinal benefits, including their unique flavor and taste. They contain low fat and abundant sources of fiber, proteins, minerals, and vitamins. Hence, these species are attracted great interest due to their medicinal properties such as antioxidant, immunomodulatory, antitumor, and antiviral.

These properties are attributed mainly to glucan and other polysaccharides (Camelini et al., 2014). Pleuran (β-glucans) is an insoluble polysaccharide isolated from Pleurotus ostreatus, which showed immunomodulation properties and even reduced the cholesterol level in humans (Rop et al., 2009; Bobek et al., 1997).

Furthermore, the roseus mushroom (P. djamor var.

roseus) is one of the attractive edible species, had been demonstrated that consist of rich source protein, fiber, amino acids, minerals, and secondary metabolites. Our previous study has reported a suitable substrate for the low-cost and high-yield production of P. djamor var.

roseus (Jegadeesh et al., 2018). In this context, present studies have focused on the functional properties of water-soluble polysaccharide extraction and purification from the basidiocarps of P. djamor var. roseus. As a result, its isolation and chemical characterization and antioxidant activities were reported for the first time. The result of this study would highlight the significance of P. djamor var.

roseus as a possible valuable source for β-d-heteroglycan, which would help to exhibit unique antioxidant properties.

MATERIALS AND METHODS

Materials and Equipment

The experimental strain of Pleurotus djamor var.

roseus, was harvested from the paddy straw substrate, collected from Centre for Advanced Studies in Botany, University of Madras (Fig. 1). Fresh basidiocarps (2 kg) were washed with tap water and oven-dried at 55^oC for 24 h. The dried samples were powdered (coarse) using a

blender. All analytical grade chemicals and reagents used in this study were purchased from Hi-Media (Bengaluru, India), Merck (Darmstadt, Germany), Sigma Chemicals (Burlington, MA, USA). Distilled and ultrapure water were obtained using a water filtration system (Millipore, Billerica, MA). Spectrophotometric measurements were performed using a PowerWave X340 Microtiter-Plate ELISA Reader (Bio-Tek Instruments, Inc., Winooski, VT, USA) and a Fourier-transform infrared (FT- IR) spectrophotometer (Perkin-Elmer, Waltham, MA, USA).

Extraction of crude polysaccharide (CPs)

The dried basidiocarps were then grounded using an electrical blender into a coarse powder (prevent bumping) and stored in airtight containers at 4^oC prior to use. The coarse powder 200 g was mixed with triple-distilled water at a ratio of 1:10 (w/v) and then boiled at 100^oC for 4 hours with regular stirring. Then the extract was cooled, the residues were removed by filtration through absorbent cotton gauze cloth and centrifuged at 10,000 g for 30 min at 4^oC. The liquid fraction was precipitated by the addition of 95% ice-cold ethanol (1:4) and kept at 4^oC overnight. The precipitate was collected by centrifugation at 10,000 g for 20 min. The supernatant was collected and concentrated using a rotary evaporator. Further, the pellet was collected, dissolved in glass distilled water, and dried to powder (labeled as CPs). The total sugar content was estimated using glucose as a standard by the phenol sulfuric acid methods (Dubois et al., 1956).

The dried CPs was treated with acetone to remove pigments, and the protein contaminants were removed by Sevag methods (Staub, 1965). Free from contaminants, the powder was dissolved in glass distilled water and exhaustively dialyzed against distilled water overnight.

Fig. 1. Cultivated Pleurotus djamor var. roseus on paddy straw substrate.

The recovered brownish CPs was subjected to a purification procedure.

Purification of P. djamor var. roseus polysaccharide The obtained CPs was further purified according to the methods described by He et al. (2010), with minor modifications. The CPs (560 mg) was loaded onto a DEAE cellulose column (52 × 2.5 cm) and eluted with ultrapure water (non-adsorbed fraction -NAF) and later with 0.2 M and 0.5 M NaCl solution (adsorbed fraction- AF), respectively. The NAF fractions were collected at the flow rate of 0.5 mL/min. Father, positive NAF fraction was purified by sepharose-6B (Sigma, USA) column 89 × 6 cm and eluted with ultrapure water at the flow rate of 0.1 mL/min. The total sugar content in each fraction was determined. The purified polysaccharide (PPs) was collected and lyophilized for further analysis.

Determination of total protein and phenol

Free proteins were determined by Bradford method (1976) using BSA equivalents of dry weight. Total phenolic contents of CPs and PPs were determined by Folin–Ciocalteu method using gallic acid as the standard (Singleton et al., 1999).

Determination of glucan content

Total glucan, α- and β-glucan in the CPs and four column factions (FA, FB, FC, F1) were determined using β-glucan assay kit K-YBGL 12/6 (Megazyme, Ireland). The experiment was performed according to the manufacturer’s protocol. The β-glucan content in the polysaccharide fraction was calculated by subtracting the amount of α-glucan from total glucan content.

Thin-layer chromatography

Thin-layer chromatography (TLC) was performed on a silica gel plate (E-Merck, Germany). To detect the monosaccharide’s present in the PPs, the sample was acid hydrolyzed with 2 M trifluoroacetic acid (TFA) for 18 h. The hydrolyzed PPs sample was spotted along with standard sugar solutions and developed using the solvent system, n-butonal: acetic acid: water (2:1:1). The spots were visualized by spraying the plate with 4- aminobenzoic acid-acetic acid-phosphoric acid reagent and kept at 100^oC for 20 min. The Rf value of the spots was calculated and compared with standard sugars (glucose, mannitol, maltose, lactose, and xylose).

Chemical characterization of P. djamor var. roseus polysaccharide

The purified polysaccharide (PPs) was chemically characterized using an FT-IR spectrometer. The FT-IR spectra were recorded at frequencies ranging from 400 to 4000 cm^-1 using a Perkin-Elmer FT-IR spectrophotometer.

In vitro antioxidant activity

The free radical-scavenging (DPPH) and superoxide radical-scavenging activities of PPs were determined according to the methods of Brand-Williams et al., (1995) and Liu et al. (1997), respectively.

Statistical analysis

All results were obtained in triplicate, and data were expressed as mean ± standard deviations. The data were subjected to a One-way Analysis of Variance (ANOVA) to determine the significance of individual differences at p < 0.05 level. Duncan’s multiple range test compared to significant means. All statistical analyses were carried out using the SPSS statistical package (SPSS, Version 25, SPSS Inc., Chicago, USA).

RESULTS AND DISCUSSION

Extraction and purification of polysaccharide In this present study, polysaccharide was extracted by hot water, followed by ethanol precipitation. The resulting, brown-colored crude polysaccharide (CPs) was lyophilized, and the yield was 5.6% dw. According to Siu et al. (2014), the brown color represents the phenolic content in the polysaccharide. However, the hot water extraction and cooking process may have increased mushroom polysaccharide yield (Radzki et al., 2016). The overall protocol for isolation and purification of polysaccharides from P. djamor var. roseus is schematically demonstrated in Fig. 2. The crude polysaccharide (CPs) was purified through DEAE cellulose ion-exchange column chromatography. Three major fractions (FA, FB & FC) were eluted with a different gradient of NaCl. The total sugar content was determined in the fractions; fraction FB (27-36) showed high sugar content, and the yield was 63.42% (Table 1). Desalted FB was further purified with the sepharose-6B gel permeable column in an aqueous medium, and the two fractions F1 and F2 were obtained (Fig. 3). Both eluted fractions were collected separately and lyophilized. While the F1 fraction was

determined as high sugar content, this fraction was considered a purified polysaccharide (PPs). We obtained

300 mg of purified polysaccharide after lyophilization, and it was white. (Fig. 2).

Physicochemical properties and chemical composition of PPs

The amount of total sugar, protein, and phenolics are presented in Table 1. The total sugar content in the order of the crude isolates and fractions were obtained, CPs (61.20%), FA (34.95%), FB (63.42%), FC (8.65%), and F1 (PPs) (78.40%) on a dry weight basis. We recorded the highest sugar value in PPs compared to the previously reported in P. ostreatus (Xia et al., 2011).

The protein content decreasing in the order of fractions separate, whereas the protein content in CPs and PPs is 12.86% and 2.62%, respectively. According to Gonzaga et al. (2005), protein and glucan-protein complex are excluded from the first fractions by some selective process. While the phenolic content in CPs, FA, FB, and FC, were 0.64, 0.33, 0.12 and 0.08%, respectively. However, the phenolic content was not detected in the F1 fraction. On the other hand, other studies demonstrated that phenolic content in purified polysaccharides was less than 1%

recorded in oyster mushrooms (Radzhi et al., 2016). In contrast, the boiling temperature (>100^oC) and the acidic environment may decrease in protein and phenolic content (Szwengiel & Stachowiak, 2016). Siu et al. (2014) reported that most mushrooms' polysaccharides are neutral and do not contain uronic acid. However, relatively high protein and phenolics content in a mushroom polysaccharide are highly beneficial as it may significantly increase antioxidant defense. Total glucan, α- and β-glucan in the crude extract and fractions are summarized in Table 1. Interestingly, the boiling process significantly increased the glucan content in the CPs. As depicted in Table 1, the total glucan content ranges from 76.85% to 2.95%, whereas the β-glucan content raged from 75.08% to 1.46%. Kanagasabapathy Fig. 2. Extraction and fractionation of polysaccharides from

P. djamor var. roseus.

Table 1. Chemical composition of polysaccharides obtained from Pleurotus djamor var. roseus.

Content (% dw)

Crude extract and Fractions

CPs FA FB FC F1 (PPs)

Total Sugar 61.20 ± 0.31^c 34.95 ± 0.43^b 63.42 ± 1.34^d 8.65 ± 0.52^a 78.40 ± 0.99^e Total Protein 12.86 ± 0.91^d 6.2 ± 0.5^c 3.6 ± 0.36^b 0.86 ± 0.08^a 2.62 ± 0.46^b

Total Phenol 0.64 ± 0.07^d 0.33 ± 0.06^c 0.12 ± 0.03^b 0.08 ± 0.01^ab 0^a

Total glucan 31.2 ± 1.33^c 14.28 ± 0.82^b 44.60 ± 0.78^d 2.95 ± 0.34^a 76.85 ± 1.45^e α-glucan 0.80 ± 0.07^a 0.46 ± 0.07^a 4.10 ± 0.8^b 0.08 ± 0.01^a 0.25 ± 0.06^a β-glucan 29.61 ± 0.72^c 11.08 ± 0.84^b 40.32 ± 0.72^d 1.46 ± 0.41^a 75.08 ± 0.81^e β-Glucan (% w/w)=total glucan (% w/w) − α glucan (% w/w). Each value is an average of three replicate, values are mean±standard deviation, the same alphabets in the row are not differ significantly by Duncan test (p<0.05).

et al. (2012) reported that α-glucan and β-glucan content in P. sajor-caju ranged between 5.4% and 80.55% (w/

w). Different studies elaborated that the polysaccharide content in the mushrooms is different among the species. In the present study, α-glucan content was recorded maximum in FB (4.10%), whereas, negligible for all other fractions (< 1%, dw). At the same time, β- glucan was recorded maximum in F1 fraction (PPs) 75.08%.

TLC profile of PPs

The monosaccharide composition of hydrolyzed PPs was determined along with standard sugar solutions (glucose, mannitol, maltose, lactose, and xylose) by thin- layer chromatography (TLC). Major pinkish spots were observed with an Rf value of 0.385, corresponding to glucose and lactose standards (Fig. 4(a)). According to Hereher et al., (2018), the TLC profile of

heteropolysaccharides isolated from Volvariella sp. showed xylose in addition to glucuronic acid. In addition, P.

pulmonarius crude polysaccharides contain glucose and xylose (Hereher et al., 2018). In this study, PPs were composed of monosaccharides, and the TLC profile matched with glucose and xylose (it may be xyloglucan). One of the best sources of β-glucan and they abundant present in the fungal polysaccharide.

While blanching and boiling process may increase the polysaccharide yield in A. bisporus (Radzki et al., 2019).

Gamar et al. (1997) reported that TLC is a sensitive method for qualitative analysis of polysaccharides. In the present study, the TLC profile, PPs from P. djamor var. roseus, might have glucan in its structure.

FT-IR spectroscopic characterization

Fourier transform infrared (FT-IR) spectra of the purified PPs was shown in Fig. 4(b). The pronounced Fig. 3. Elution profile of polysaccharide in DEAE cellulose and Sepharose 6B chromatography (a & b) Elutes were analyzed by measuring the absorbance at 490 nm for carbohydrate.

FT-IR bands were assigned to carbohydrates and other constituents. Based on characteristic bands of PPs, it was found that polysaccharides predominated. The typical prominent broad stretching peak at 3409 cm^-1 for the hydroxyl group and a weak band at 2926 cm^-1 showing the CH₂ asymmetric vibration, pyran. The moderately intense peak at 2920 cm^-1 corresponded to C single bond H stretching and bending vibrations of an alkyl group (Liu et al., 2015). On the other hand, other studies demonstrated that the vibrations at around 3420 and 2920 cm^‒1 are attributed to the hydroxyl stretching and C–H vibration absorption of the polysaccharide, respectively (Sermwittayawong et al., 2018). The strong

absorption peak at around 1650 cm^-1, 1638 cm^-1, and a weak one at near 1406 cm^-1 were indicative of the presence of carboxyl groups. Bands between ~1450 and 1300 cm^-1 correspond to bending vibrations of C-OH, O-H, and C-OH, CH₂ wagging (Socrates, 2001). Similarly, the C-O-C stretching vibration of the pentacyclic group of pyranoid sugar had an adsorption band at 1077 cm^-1 (Figueiro et al., 2004). C-O-H deformation vibration had a peak at 1043 cm^-1, and 885 cm^-1 was ascribed to β- pyranoses in the polysaccharide. These observations further confirmed that the PPs was composed of polysaccharide, protein, and uronic acids. Region of 950- 1190 cm^-1 contains intense absorption peaks due to stretching Fig. 4. Thin-layer chromatography (A) and B. We have revised the FTIR image please updated accordingly (B) of partially purified polysaccharide (PPs). A. (a) PPs (b) glucose (c), mannitol (d), maltose (e) xylose, (f) lactose.

vibrations of C–O–C, COH, and C–C. Bands at

~930 cm^-1, 840-860 cm^-1, and ~760 cm^-1 can be associated with the presence of a-glycosidic links (Wiater et al., 2011). According to Galichet et al. (2001), the vibrations at 800 cm^‒1 and 910-914 cm^-1, which are signatures of mannan, suggest the presence of such polysaccharides in the sample. The spectral details confirmed the presence of a polysaccharide moiety with xylopentose type of compound (Fig. 4(a)).

In vitro antioxidant activity

Mushroom-derived polysaccharides are associated with antioxidant, anti-inflammatory, and immunomodulatory activities. In addition, edible and medicinal mushrooms polysaccharides exhibited the most potent radical scavenging ability for hydroxyl radicals and inhibited tumor cell proliferation (Chen et al., 2015, Zeng et al., 2018). Due to the changes in lifestyle and increased risk of oxidative stress due to the reactive oxygen species (ROS) that damage DNA and essential proteins. The functional properties of mushroom polysaccharides ability to destroy ROS and decrease the damaging effect in our bodies. Moreover, mushroom polysaccharides containing high uronic acid levels display significant antioxidant activity and may be attributed to the functional group -COOH (Li et al., 2016). Functional groups that act as efficient electron or hydrogen donors are associated with the antioxidant activity of specific polysaccharides. These functional properties may use in various food formulations as an ingredient with enhanced antioxidants (Khan et al., 2015). However, many studies revealed that the polysaccharide derived from oyster mushrooms showed strong DPPH radical and superoxide anion radical scavenging activity (Piska et al., 2017, Rodrigues Barbosa et al., 2020). In the present study, free radical scavenging (DPPH) assay of the purified PPs derived from P. djamor var. roseus showed 50% radical scavenging activities at 2.44 mg/

mL, whereas the standard ascorbic acid showed 50%

scavenging at 0.87 mg/mL (Figure 5). It was observed that an increase in the polysaccharide concentration increased the activity level. The maximum radical scavenging activity was recorded at 5 mg/mL (82.32±2.60%), and the lowest at 0.5 mg/mL (17.62±3.47%) were recorded.

However, the DPPH radical scavenging activity of PPs was observed increasing in a dose-dependent manner.

The superoxide anion (O₂^-) is also known to indirectly initiate lipid peroxidation because of the formation of

singlet oxygen and hydroxyl radicals. Furthermore, superoxide anion radicals can magnify the cellular damage and contribute to tissue damage and various diseases (Ozsoy et al., 2009). Besides superoxide radical being a weak oxidant, it can damage the DNA and cells, leading to various diseases (Benov, 2001, Wong et al., 2007, Akhigbe et al., 2021). Even though the anion radical and its derivatives could induce oxidative damage in lipids, proteins, and DNA, it is extremely harmful to human living. These results clearly showed that the antioxidant activity of PPs related to the ability to scavenge superoxide radicals, and it was concentration dependent. In the present study the scavenging effect of the superoxide radical was 20.21±1.57%-80.3±1.83% at 0.5-5 mg/mL (Fig. 5). On the other hand, the standards L-ascorbic acid showed the scavenging effects of 83.39±3.71% at 2.5 mg/mL concentration. Most of the antitumor active polysaccharides were extracted from the fruiting body of Pleurotus species (Liu et al., 2015, Assis et al., 2013, Patel et al., 2012). Consequently, Pleurotus species may provide a wide range of Fig. 5. In vitro antioxidant activity of isolated polysaccharide from P. djamor var. roseus, (a) DPPH free radical-scavenging, (b) superoxide radical-scavenging activity.

polysaccharides, and they are excellent metabolites that have revealed important biological activities. The results showed that DPPH and superoxide radical-scavenging activity PPs increase gradually as the concentration of polysaccharides increases from 0.5 to 5.0 mg/mL. On the other hand, other studies demonstrated that polysaccharides from Pleurotus ostreatus have shown similar antioxidant activity (Uddin et al., 2019). The results revealed that antioxidant activity of PPs is low compared to that of vitamin C below 0.87 mg/mL and in case of superoxide radical scavenging that concentration is 1.25 mg/mL, (P<0.05).

CONCLUSION

The presence of polysaccharides in P. djamor var.

roseus attributes to the antioxidant and anticancer activity. The mushroom-derived polysaccharides exhibit low toxicity to mammals and display numerous pharmacological properties. Pleurotus species are commercially important mushrooms, and they are widely cultivated throughout the world. They are an excellent source of nutrient and bioactive substances. The purified polysaccharide (PPs) from P. djamor var. roseus is composed of monosaccharides and a small number of proteins. The results exhibited intense activity on free radical and superoxide anion radical scavenging activity. At the same time, the purpose of this study was to explore and provide an opportunity to obtain navel pharmacological agents from P. djamor var. roseus. Hence, PPs might be used as a natural antioxidant in the pharmacological industry.

However, further investigations are required to investigate the immunomodulation and antitumor properties of PPs and to develop active ingredients.

ACKNOWLEDGEMENT

The first author thanks the Department of Chemical Engineering, Graduate School of Chosun University and Korea Forest Service (Korea Forestry Promotion Institute - Project no. 2020191B10-2022-BA01), Republic of Korea, for the postdoctoral fellowships.

CONFLICT OF INTEREST

Authors declare that they do not have any conflict of

interests.

적 요

버섯 다당류는 면역 조절 기능, 항암 및 항산화 활성을 비롯하여 항바이러스 활성과 방사선 스트레스의 경감 등 인 체에 유익한 다양한 생리활성을 나타낸다. 본 연구에서는 분홍느타리버섯 (Pleurotus djamor var. roseus Corner)으 로부터 뜨거운 물과 에탄올 침전을 이용하여 5.6%의 수율 로 갈색을 띠는 경화된 다당류(CPs)를 순차적으로 추출하 였다. 이렇게 추출된 CP는 Diethylaminoethyl cellulose (DEAE) 및 sepharose-6B 컬럼 분리를 통해 4개의 분획을 얻었고 각 분획의 총 글루칸 함량은 각각 76.85%, 2.95%, 75.08%, 1.46%로 밝혀졌다. 이중 가장 높은 수율의 분획 (PP)으로부터 300 mg의 백색 분말이 얻어 졌으며, 박층 크 로마토그래피(TLC)와 푸리에 변환 적외선 분광법(FTIR) 의 결과로부터 자일로펜토스 유형의 화합물과 함께 다당류 부분의 존재를 확인하였다. PP의 항산화 활성은 1,1- diphenyl-2-picryl-hydrazyl(DPPH) 자유라디칼 소거 분석 및 슈퍼옥사이드 라디칼 소거 분석을 통하여 높은 활성을 나타냄을 확인하였다. PP분획에는 페놀, 단백질 및 단순 탄 수화물이 없는 정제된 베타글루칸이 주 구성성분으로, 정 제된 다당류가 천연 항산화제로 사용될 수 있음을 알 수 있 었다.

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