review article natural products and biological...

Hindawi Publishing CorporationBioMed Research InternationalVolume 2013 Article ID 982317 9 pageshttpdxdoiorg1011552013982317

Review ArticleNatural Products and Biological Activity of thePharmacologically Active Cauliflower MushroomSparassis crispa

Takashi Kimura

Research amp Development Center Unitika Ltd 23 Uji-Kozakura Uji Kyoto 611-0021 Japan

Correspondence should be addressed to Takashi Kimura takashi-kimuraunitikacojp

Received 4 October 2012 Accepted 25 February 2013

Academic Editor Fabio Ferreira Perazzo

Copyright copy 2013 Takashi Kimura This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

Sparassis crispa also known as cauliflower mushroom is an edible mushroom with medicinal properties Its cultivation becamepopular in Japan about 10 years ago a phenomenon that has been attributed not only to the quality of its taste but also toits potential for therapeutic applications Herein I present a comprehensive summary of the pharmacological activities andmechanisms of action of its bioactive components such as beta-glucan and other physiologically active substances In particularthe immunomodulatory mechanisms of the beta-glucan components are presented herein in detail

1 Introduction

Medicinal mushrooms have an established history of use intraditional Asian therapies Over the past 2 to 3 decadesscientific and medical research in Japan China and Koreaand more recently in the United States has increasinglydemonstrated the potent and unique properties of com-pounds extracted from mushrooms for the prevention andtreatment of cancer and other chronic diseases Variousimportant pharmaceutical products with proven medicinalapplications have been derived from mushrooms [1]

Sparassis crispa WulfFr (Figure 1) also known ascauliflower mushroom is an edible mushroom with variousmedicinal properties whose cultivation has recently becomepopular in JapanThe taxonomyof S crispa is as follows king-dom Fungi phylum Basidiomycota class Agaricomycetesorder Polyporales family Sparassidaceae genus Sparassisand species crispa It is a brown-rot fungus that primarilygrows on the stumps of coniferous trees and is widelydistributed throughout the North Temperate Zone S crispahas been reported to have many biological activities whichare detailed below

2 Chemical Constituents and BioactiveComponents of S crispa

Scientific investigation has led to the isolation of many com-pounds from S crispa that have been shown to have health-promoting activities The fruiting bodies of S crispa containapproximately 90 water protein lipid carbohydrate ashand dietary fiber (Table 1) [3] Furthermore the content ofvitamin D

2 which aids intestinal calcium absorption was

shown to be 017mg per 100 g of dry weight a concentrationthat is higher than that observed in other mushrooms [4]Also S crispa contained a relatively large amount of glucosylceramide (approximately 02) which is a glycoside ofceramide It was demonstrated that the moiety of sphingoidbase was characterized by the unique structure [5] ThoughS crispa has a scent of its own the results of headspaceanalyses showed that 3-octanone DL-3-octanol and 1-octen-3-ol contributed mutually to the particular aroma of thismushroom [6] It is noteworthy that the beta-glucan contentof S crispa is more than 40 of the dry weight of the fruitingbodies asmeasured by the enzymemethod of the Japan FoodResearch Laboratories (Tokyo) [3]

2 BioMed Research International

10 cm

Figure 1 Sparassis crispaWulfFr [2]

Table 1 Approximate composition of Sparassis crispa (per 100 g drysample)

Components Amount (g)Protein 134Fat 20Ash 18Carbohydrate 215Dietary fiber (DF) 612Beta-glucan from DF 435

21 Polysaccharide (Beta-Glucan)

211 Primary Structure Using chemical enzymatic andNMR analyses it was shown that the primary structureof a purified beta-glucan (designated SCG) obtained fromcultured fruiting bodies of S crispa is a 6-branched 13-beta-glucan with one branch in approximately every 3 main chainunits (Figure 2) [7ndash9]

212 Biological Activities Tumor size in cancerous (Sarcoma180) ICR mice was dose-dependently decreased after 5 weeksof oral administration of S crispa (10 or 100mgkg) incomparison to a control group Furthermore the survival rateof these model mice was higher when similarly treated withS crispa [2] Since SCG content in dry powder of S crispawasmeasured to bemore than 40 SCGwas likely be responsiblefor this antitumor effect

Ohno et al prepared polysaccharide fractions from thefruiting bodies of cultured S crispa and showed their anti-tumor activity against the solid form of Sarcoma 180 in ICRmice with strong vascular dilation and hemorrhage reactions[7] Furthermore intraperitoneal and oral SCG over a widerange of concentrations enhanced hematopoietic responsesin mice with leukopenia induced by cyclophosphamide (CYa DNA-alkylating agent) [10 11] This effect was augmentedin combination with isoflavone aglycone [12] SCG was also

shown to stimulate leukocytes to produce cytokines such asIL-8 in whole-cell cultures of human peripheral blood [13]and in mouse splenocytes [14]

Yamamoto et al reported antiangiogenic and anti-metastatic effects of SCG on neoplasm by using differentanimal models [8] Oral administration of SCG suppressedB16-F10 cell-induced angiogenesis in a dorsal air sac assayusing ICR mice and suppressed vascular endothelial growthfactor induced neovascularization in a Matrigel plug assayusing C57BL6J mice Furthermore it suppressed the growthand number of metastatic tumor foci in the lung alongwith primary tumor growth in a C57BL6J mice model ofspontaneous metastasis From these findings it is apparentthat the oral administration of SCG exerts a suppressive effecton tumor growth and metastasis in the lung through theinhibition of tumor-induced angiogenesis

Taken together these results demonstrate that SCG ex-hibits various biological activities including antitumoreffects enhancement of the hematopoietic response andinduction of cytokine production in vivo and in vitro

213 Mechanisms Harada et al reported strain-specificdifferences of the reactivity of mice to SCG with DBA1 andDBA2 mice being highly sensitive to SCG In splenocytesderived from various inbred strains of mice interferon-120574(IFN-120574) production was not induced by SCG Howeversplenocytes from naıve DBA1 and DBA2 mice stronglyreact with SCG to produce IFN-120574 [14] Furthermorein addition to IFN-120574 cytokines induced by SCG werescreened for and found to include tumor necrosis factor-120572(TNF-120572) granulocyte-macrophage colony-stimulating factor(GM-CSF) and interleukin-12 (IL-12p70) [15] Since the seraof naıve DBA1 and DBA2 mice contain significantly highertiters of antibody against SCG than other strains of mice [16]it seems likely that these mice strains are sensitive to SCGThus DBA1 and DBA2 mice would be useful models forfuture studies of SCG

Harada et al further demonstrated that GM-CSF was oneof the key factors in reactivity to SCG in DBA2 mice [15 17]Neutralizing GM-CSF using an anti-GM-CSF monoclonalantibody significantly inhibited IFN-120574 TNF-120572 and IL-12p70elicited by SCG The splenocytes in various strains of miceshowed similar patterns of cytokine production in responseto SCG cotreatment in the presence of recombinant murineGM-CSF The high sensitivity to SGG shown by DBA1and DBA2 mice may be attributable to differences of theirregulation of GM-CSF compared with that in other mice

Harada and Ohno also proposed an interesting model forthe mechanism of cytokine induction by SCG in DBA2mice[18] Broadly speaking SCG directly induces adherent cellsto produce TNF-120572 and IL-12p70 whereas cell-cell contactmediated by the association of CD4+ T cells expressing LFA-1and antigen-presenting cells such as dendritic cells expressingICAM-1 is required for the induction of IFN-120574 and GM-CSFby SCG

Neutrophils macrophages and dendritic cells expressseveral receptors capable of recognizing beta-glucan in its

BioMed Research International 3

O

H

HO

H

O

H

H

OHH

OH

O

H

H

H

HOHH

O

H

HO

H

H

H

OHHO

OH

O

O

HHO

H

HO

H

H

OH

H

O

OH

OH

119899

O

Figure 2 Chemical structure of SCG [9]

various forms Dectin-1 complement receptor 3 lactosylce-ramide and scavenger and Toll-like receptors are all candi-dates that have been reported thus far [19ndash23] Among thesedectin-1 which is a C-type lectin is an archetypical non-Toll-like pattern recognition receptor expressed predominantlyby myeloid cells Dectin-1 can induce its own intracellularsignaling and canmediate a variety of cellular responses suchas cytokine production [24]

The magnitude of cytokine induction from bone-marrow-derived dendritic cells (BMDCs) by SCG and theexpression level of dectin-1 on BMDCs in DBA2 mice areboth higher than that of other strains of mice Furthermoreblocking dectin-1 significantly inhibits the induction ofTNF-120572 production by SCG These results suggest that theBMDCs from DBA2 mice are highly sensitive to SCG-induced cytokine production in vitro and that this sensitivityis related to the expression level of dectin-1 [25]

The molecular mechanism of the enhanced hematopoi-etic response has been investigated in CY-treated mice (bothICR and C57BL6 strains) [26] According to this reportthe levels of IFN-120574 TNF-120572 GM-CSF IL-6 and IL-12p70were all shown to be significantly increased in SCG-treatedsplenocytes of CY-treated mice GM-CSF production inthe splenocytes of CY-treated mice was reportedly higherthan that in normal mice regardless of SCG stimulationNeutralizingGM-CSF significantly inhibited the induction ofIFN-120574 TNF-120572 and IL-12p70 by SCG The level of cytokineinduction by SCG was modulated by the amount of endoge-nous GM-CSF produced in response to CY treatment ina dose-dependent manner The expression of beta-glucanreceptors such as CR3 and dectin-1 was upregulated byCY treatment Blocking dectin-1 significantly inhibited theinduction of TNF-120572 and IL-12p70 production by SCG Takentogether these results suggest that the key factors in cytokineinduction in CY-treated mice are the enhanced levels of bothendogenous GM-CSF production and dectin-1 expressionIt is very interesting that agents that modulate GM-CSF

production and dectin-1 expression such as CY can controlreactivity to SCG and the expression of various cytokines

Shibata et al found that both GM-CSF and TNF-120572synthesis in DBA2 mouse splenocytes stimulated with SCGbut not with lipopolysaccharide were significantly enhancedin the presence of cytochalasin D (CytD) an inhibitor ofactin polymerization [27] On the other hand Kim et alpointed out the importance of the role of Toll-like recep-tor 4 (TLR4) [28] They examined the effect of SCG onadherent monocytes such as macrophages and dendriticcells (DCs) and nonadherent lymphocytes such as T and Bcells and demonstrated that SCG mainly activated DCs andmacrophages but not T and B cells The role of TLR4 as amembrane receptor of SCG was shown by the impairmentof maturation of DCs generated from bone marrow cells oftlr4minusminus knockout mice and TLR4-mutated C3HHeJ miceand by using an anti-MD-2TLR4 neutralizing antibody SCGincreased the phosphorylation of ERK p38 and JNK andenhanced nuclear translocation of NF-120581B p50p65 in DCsThese results indicate that SCG activates DCs via MAPK andNF-120581B signaling pathways which are signaling moleculesdownstream of TLR4

22 Low-Molecular-Weight Compounds S crispa possesses awide range of bioactive metabolites which are products ofsecondary metabolism (Figure 3)

221 Antimicrobial Compounds It has been reported thatS crispa produces antibiotic substances For example sup-pression of Bacillus subtilis growth on agar media is knownto be due to sparassol (methyl-2-hydroxy-4-methoxy-6-methylbenzoate) (1) [29] Woodward et al reported that Scrispa produced 3 antifungal compounds when submergedin culture in a 2 malt broth The compounds included


sparassol and two other antifungal compounds methyl-24-dihydroxy-6-methylbenzoate (2) and methyl-dihydroxy-methoxy-methylbenzoate (the positions of substituents wereunclear) both of which showed higher antifungal activitythan sparassol against Cladosporium cucumerinum [30]

A novel compound (4) and a previously known one (3)were isolated from S crispa [31] Both compounds wereshown to inhibit both melanin synthesis and methicillin-resistant Staphylococcus aureus (MRSA) growth The mini-mum inhibitory concentration (MIC) values of compounds3 and 4 in the anti-MRSA assay were 05 and 10mMrespectively IC

50values of compounds (3) and (4) in the

melanin production inhibition assay were 33120583M and 12 120583Mrespectively Since the IC

50value of an existing whitening

agent arbutin is reported to be 132mM these compoundshave potential as constituents of cosmetic products

In the course of screening for compounds that inhibitMRSA growth Kodani et al discovered 2 known chalconesxanthoangelol (5) and 4-hydroxyderricin (6) in the extractof S crispa which have been previously isolated from theplant Angelica keiskei These compounds showed anti-MRSAactivity and their MICs were 2 and 025mM respectivelyThis was the first report of the isolation of chalcones from arepresentative of the Fungi kingdom [32]

222 Other Bioactive Compounds A new sesquiterpenoidwas also isolated from S crispa [33] Its structure wasdetermined to be (3Rlowast 3aSlowast 4Slowast 8aRlowast)-3-(11015840-hydroxy-11015840-methylethyl)-58a-dimethyldecahydroazulen-4-ol (7) by acombination of NMR and ESI-MS analyses This was thefirst isolation of an isodaucane-type sesquiterpenoid from afungus including mushrooms

Yoshikawa et al isolated three novel phthalides desig-nated hanabiratakelide A (8) B (9) and C (10) in addition tothree known phthalides from the S crispa fruiting body [34]The 6 isolated compounds were tested for their antioxidantactivity The in vitro superoxide dismutase-like activity ofthe three hanabiratakelides was stronger than that of vita-min C These compounds also inhibited lipopolysaccharide-stimulated nitric oxide and prostaglandin E2 production bya murine macrophage cell line RAW264 In addition thegrowth of the colon cancer cell lines Caco-2 and colon-26was significantly inhibited by treatment with all 3 of thehanabiratakelides

3 Pharmacological Aspects of S crispa

31 Antiviral Activity Reverse transcriptase (RT) is one ofthe key enzymes in human immunodeficiency virus (HIV)replication HIV replication is interfered with when theenzyme is inhibited Thus RT inhibitors can be used to treatAIDS Hot water extracts from the fruiting bodies of 16species of mushroom including S crispa were screened forHIV-1 RT inhibitory activity The extract of S crispa elicited703 inhibition when tested at a concentration of 1mgmLHowever the active component remains unclear [35]

32 Antihypertensive Effects One of the main causes ofstroke is hypertension Therefore it is important to avoid

high blood pressure as a preventative measure Yoshitomi etal investigated not only the preventive effects of S crispaagainst stroke and hypertension in stroke-prone sponta-neously hypertensive rats (SHRSP) but also the mechanisminvolved by studying the cerebral cortex [36] SHRSP ratsgiven feed containing 15 S crispahad a delayed incidence ofstroke and death significantly decreased blood pressure andincreased blood flow Moreover the urinary nitratenitriteexcretion and the nitratenitrite concentration in cerebraltissue were higher than those of control SHRSP rats In thecerebral cortex phosphor-eNOS (Ser1177) and phosphor-Akt (Ser473) in S crispa-treated SHRSP rats were increasedcompared with those of control SHRSP rats In conclusionS crispa can ameliorate cerebrovascular endothelial dysfunc-tion by promoting recovery of Akt-dependent eNOS phos-phorylation and increasing nitric oxide (NO) production inthe cerebral cortex

In addition Lee et al indicated that SCG was able tostimulate NO production as well as enhance the expres-sion of inducible NO synthase (iNOS) from macrophage-like RAW2647 cells [37] Since NO production is stronglysuppressed by mitogen-activated protein kinase (MAPK)inhibitors it is likely that SGG-induced NO release is medi-ated by MAPK

33 Antidiabetic Activity It has been shown that dietary Scrispa improves the symptoms of both type 1 and type 2diabetes The consumption of a diet containing more than05 S crispa results in significant improvement in diabetessymptoms (body weight loss and increased blood glucose)in ICR mice with STZ-induced diabetes [38] FurthermoreYamamoto and Kimura examined the effect of dietary Scrispa on KK-Ay mice an animal model of type 2 diabetesmellitus [39] The group that was fed 50 S crispa dietshowed not only a significant decrease of blood glucose andinsulin levels but also a pronounced increase in plasmalevels of adiponectin in comparison with a control groupAlthough the S crispa diet had no effect on body and adiposetissue weights in KK-Ay mice the size of the mesentericadipose cells of mice in the S crispa group tended to besmaller than the control group Thus the S crispa dietmay decrease the adipose cell size in order to increaseplasma adiponectin levels Considering the physiologicalsignificance of adiponectin these findings imply that dietaryS crispa has the potential to ameliorate type 2 diabetes

GPR40 is one of the G protein-coupled receptors whichhas 7 transmembrane spanning helical bundles GPR40distributes in pancreas and central nervous system It can bebound by medium- and long-chain fatty acid and activatethe intracellular signal pathways which in turn regulates thefunction of cells In pancreatic beta-cell intracellular calciumconcentration elevates when GPR40 is binding to fatty acidthereby promoting the release of insulin [40] Yoshikawa etal demonstrated that a couple of unsaturated fatty acids in Scrispa were the agonist of GPR40 which might be used forpreventing and treating the diabetes [41]

The normal healing process in healthy individuals takesplace at an optimal rate but it is usually delayed or even


N

OH

OO

O

N

OH

OO

O

OHOH

H

HO OH

OH O

OH

OH O

CH3O

O

OH

HO

O

O

HO

OOH

O

HO

HOO

OH

OH O

HO

OH O

CH3O CH3

OCH3 OCH3

CH3

OCH3

CH3OCH3O

(1) (2)

(3) (4)

(5)

(6) (7)

(8) (9) (10)

Figure 3 Chemical structure of the low-molecular-weight compounds found in S crispa [29ndash34]

completely impaired in patients with diabetes Thus theimpaired wound healing that occurs in diabetes mellitus isa major clinical problem It is also generally accepted thatwound repair is an immune-mediated physiologic mecha-nism Oral administration of 1000mgkg body weight perday of S crispa for 4 weeks was shown to significantly

accelerate wound healing in rats with streptozotocin- (STZ-)induced diabetes which is an insulin-dependent model ofdiabetes mellitus (type 1) [42] Furthermore in S crispa-treatedwounds therewere significant increase inmacrophageand fibroblast migration collagen regeneration and epithe-lialization compared with a control group Therefore the use


of S crispamay be extended to the clinical setting and it mayeffectively promote wound healing in patients with diabetes

Yamamoto and Kimura investigated whether oral andtopical administration of S crispa could restore effectivewound healing in ICR mice with STZ-induced diabetes [38]Mice consuming a diet containing more than 05 S crispashowed significantly improved wound healing Notably therate of wound healing in mice fed a diet containing 25S crispa was almost the same as that in mice treated withtopical trafermin (basic fibroblast growth factor formula-tion) Moreover topically administered SCG significantlypromoted wound healing in mice with diabetes resulting ina wound contraction ratio of 37 after treatment for 9 daysa result that was superior to that of trafermin

34 Antitumor and Anticarcinogenic Activity (except for SCG)Yamamoto et al investigated the antitumor effects of alow-molecular-weight (below approximately 8 kDa) fraction(FHL) containing no beta-glucan isolated from a hot waterextract of S crispa [43] The oral administration of FHL(30mgkg) to tumor- (sarcoma 180) bearing ICR mice wasobserved to suppress tumor growth Furthermore the IFN-120574level in the culture supernatant of splenic lymphocytes fromFHL-fed tumor-bearing mice was significantly increasedcompared to a control group Tumor-induced angiogenesisin the dorsal air sac (DAS) system was also suppressedby FHL administration These results suggest that the oraladministration of FHL induces antitumor activity throughthe enhancement of the Th1-response in tumor-bearingmice Additionally the antiangiogenic activity of FHL maycontribute to its antitumor activity

Yoshikawa et al investigated the possible preventiveeffects of S crispa on azoxymethane-induced colon aberrantcrypt foci (ACF) in F344N rats S crispa feeding dose-dependently suppressed the malignant changes of ACF by54 (03 group) 64 (10 group) and 75 (30 group)They concluded that the anticancer-related activity mayoriginate from the aforementioned hanabiratakelides [34]

35 Antiallergic Activity Allergic inflammatory diseasessuch as food allergy asthma hay fever and atopic dermatitisare increasing worldwide Some recent reports have demon-strated antiallergic activities of S crispa Atopic dermatitis(AD) is a common inflammatory skin disease for whichfew effective treatments are available Oral administration of250mgkg body weight per day of S crispa decreased bothblood immunoglobulin E (IgE) level and scratching indexin NcNga mice with dermatitis induced by a continuousapplication of 246-trinitrochlorobenzene [2] In additionthe antirhinitis properties of S crispa were also investigatedin mice [44] To determine the immunomodulatory activityof oral S crispa splenocytes obtained from ovalbumin-sensitized BALBc mice fed 025 S crispawere restimulatedin vitro with the same antigen The oral S crispa inducedIFN-120574 secretion but inhibited IL-4 and IL-5 secretion andsuppressed ovalbumin-specific IgE secretion by the spleno-cytes The effects of S crispa were further investigated by

using an allergic rhinitis model in BALBc mice Nasal symp-toms sneezing and nasal rubbing induced by ovalbuminchallenges were inhibited by oral administration of S crispa(36 or 120mgkg) in a dose-dependentmanner Furthermoreovalbumin-specific serum IgE levels were diminished by Scrispa treatment in this model These results demonstratethat S crispa may be effective in suppressing symptoms ofallergic rhinitis through suppression of theTh2-type immuneresponse

Kim et al reported the effect of a water extract of Scrispa (WESC) on mast-cell-mediated allergic inflammationand the possible mechanisms of action using in vivo andin vitro models [45] WESC inhibited compound 4880-induced systemic anaphylaxis and serum histamine releasein mice WESC decreased IgE-mediated passive cutaneousanaphylaxis Additionally WESC reduced histamine releaseand intracellular calcium in human mast cells activatedby both phorbol 12-myristate 13-acetate (PMA) and cal-cium ionophore A23187 Since intracellular calcium playsan important role in the release of histamine and theexpression of cytokines the decreased intracellular calciumlevels may be involved in the inhibitory effect of WESCon histamine release WESC decreased PMA and A23187-stimulated expression of proinflammatory cytokines suchas TNF-120572 interleukin- (IL-6) and IL-1120573 The inhibitoryeffect of WESC on proinflammatory cytokines was shownto be dependent on nuclear factor-120581B extracellular signal-regulated kinase and p38 mitogen-activated protein kinaseSince the beta-glucan content in WESC was measured tobe 393 beta-glucan may be responsible for its antiallergiceffects

4 Human Clinical EvaluationIn a study where healthy men were given S crispa powderorally at 300mg per day for 8 weeks NK cell cytotoxicitywas significantly enhanced without increasing the numberof NK cells when compared to preadministration [2] Inaddition Kimura investigated whether dietary S crispainfluenced human skin condition [46] Oral administrationof S crispa powder (320mgday) for 28 consecutive daysdramatically reduced transepidermal water loss an indicatorof the skin barrier condition while that of a placebo groupwas unchanged during the testing periodThese observationsimply that oral administration of S crispa has a positive effecton the skin barrier

A clinical trial of S crispa used an orally delivered powder(300mgday) in patients with several different types of cancer(lung stomach colon breast ovarian uterine prostatepancreas and liver cancers) after the patients had receiveda single course of lymphocyte transfer immunotherapy [47]Patient assessment of 14 cases after a several month follow-up period (mean 15 months) revealed that the performancestatus in 9 cases showed improvement in quality of life andso forth

5 Conclusions and Future Prospects

S crispa has been described in the literature as a mush-room with great potential for therapeutic applications The


medicinal value of this mushroom is mainly attributable toits abundant 6-branched 13-beta-glucan (SCG) By chemicalanalysis we found that the primary structure of a purifiedbeta-glucan obtained from liquid cultured mycelium of Scrispa was a 6-branched 13-beta-glucan having one branchapproximately every 6 residues with a degree of branchingthat is relatively less than that of SCG The effect on tumor-(sarcoma 180) bearing ICR mice was much weaker than thatof SCG given by oral administration (data not shown) Fur-thermore using 1D- and 2D-NMR spectroscopy Tada et alelucidated the fine primary structure of SCG and comparedit with sonifilan (SPG) from Schizophyllum commune whichis a 6-branched 13-beta-glucan and has been used clinicallyfor cancer therapy in Japan examining differences in thebiological effects between these beta-glucans [9] Thoughboth major structural units are the same beta-(1ndash3)-glucanbackbone with single beta-(1ndash6)-glucosyl side branchingunits every 3 residues the production of IL-6 and TNF-alphafrom BMDCs was significantly increased by SCG whereasthese effects were not observed with SPG treatment Thesefindings may indicate that the biological activities of beta-glucan are attributable not only to its primary structure butalso to its conformation

Though it has been suggested that dietary S crispais useful for cancer immunotherapy in combination withlymphocyte transplantation [47] the study described in thisprevious report did not include a randomized control groupFurther clinical trials are needed to confirm the pharmaco-logical activity of dietary S crispa There is still little scientificevidence to explain the differences in responsiveness to beta-glucan in humans The studies of differences in reactivity toSCG in different animal strains [14 15 17] are important fromthis viewpoint Furthermore it is interesting that agents suchas CY can control reactivity to SCG as well as the expressionof various cytokines [26] Further research on reactivity toSCG could provide clues for developingmore effective cancerimmunotherapies using SCG

As mentioned above dectin-1 and TLR4 have been pro-posed as SCG receptors It is noteworthy that either treatmentwith a blocking antibody against dectin-1 [25 26] or geneticdeletion of TLR4 [28] completely prevents SCG-inducedDC maturation These observations might indicate that onesignaling pathway did not compensate for the other in SCG-treated DCs suggesting that both receptors are required forSCG action However further analysis of the role of thesereceptor candidates which contain complement receptor 3lactosylceramide and scavenger-like receptors in responseto SCG would be needed in order to clarify the details of itsmechanism of action in DCs [48]

The question has been raised as to how orally admin-istered beta-glucan exerts its effects The evidence pre-sented in this review clearly indicates that dietary SCG hasimmunomodulatory actions Therefore it must be assumedthat orally ingested SCG interacts with either intestinalepithelial cells andor intestinal DCs ultimately resulting inthe priming or activation of other immune cells

The antitumor mechanisms of SCG except for itsimmunomodulatory actions have not been well studiedHence we tried to elucidate the possible mechanisms of its

antiangiogenic effects As a result it was demonstrated thatSCG has both antiangiogenic functions and antimetastaticeffects on neoplasm using different animal models [8] Theantitumor effects of SCG may be partially attributable toits antiangiogenic actions Numerous reports concerning theantitumor activity of ediblemushrooms have taken particularnotice of beta-glucan However few studies have focused onantitumor components other than beta-glucan It is worthmentioning that S crispa has been shown to produce somelow-molecular-weight constituents with antitumor activitysuch as hanabiratakelide [34] and FHL [43]

Recently Park et al reported a novel process for nanopar-ticle extraction of beta-D-glucan from S crispa using insol-uble tungsten carbide [49] This nanoknife method results inhigh yields of SCG (702) with an average particle size of 150and 390 nm The extracted SCG showed a remarkably highwater solubility of 90 at room temperature This nanoknifemethod could be a potent technology to produce SCG forfood cosmetics and pharmaceutical industries

The extract of S crispamight be applied to produce healthproducts such as food beverage and antineoplastic drugActually S crispa extractions resveratrol and collagen pep-tide were claimed as antiaging agents and food supplements[50] Formulation examples of granules and health drinkswere disclosed

Many people in Japan consume S crispa and to date noreports of adverse events due to S crispa consumption havebeen reported Therefore dietary treatment with S crispamay prove to be a safe therapy for cancer and other chronicdiseases

References

[1] S PWasser andA LWeiss ldquoMedicinal properties of substancesoccurring in higher basidomycetes mushrooms current per-spectivesrdquo International Journal of Medicinal Mushrooms vol1 pp 31ndash62 1999

[2] A Hasegawa M Yamada M Dombo R Fukushima NMatsuura and A Sugitachi ldquoSparassis crispa as biologicalresponse modifierrdquo Gan To Kagaku Ryoho vol 31 no 11 pp1761ndash1763 2004

[3] T Kimura and M Dombo ldquoSparassis crispardquo in BiologicalActivities and Functions of Mushrooms H Kawagishi Ed pp167ndash178 CMC Press Tokyo Japan 2005

[4] H J Shin D S Oh H D Lee H B Kang C W Lee and W SCha ldquoAnalysis of mineral amino acid and vitamin contents offruiting body of Sparassis crispardquo in Journal of Life Sciences vol17 pp 1290ndash1293 2007

[5] M Dombo K Matumura K Yuki and T Kimura ldquoCeramidesfrom Sparassis crispardquo Japanese Kokai Tokkyo Koho P2005-206576A (August 2005)

[6] Q Wang J Li Y Wang X Zhang and K Li ldquoComparison ofaromatic components from Sparassis crispa extracted by StaticHeadspace and Headspace-SPMErdquo Shipin Gongye Keji vol 32no 11 pp 174ndash176 2011

[7] N Ohno N N Miura M Nakajima and T Yadomae ldquoAntitu-mor 13-120573-glucan from cultured fruit body of Sparassis crispardquoBiological and Pharmaceutical Bulletin vol 23 no 7 pp 866ndash872 2000


[8] K Yamamoto T Kimura A Sugitachi and N MatsuuraldquoAnti-angiogenic and anti-metastatic effects of 120573-13-D-glucanpurified from hanabiratake Sparassis crispardquo Biological andPharmaceutical Bulletin vol 32 no 2 pp 259ndash263 2009

[9] R Tada THaradaNNagi-Miura et al ldquoNMRcharacterizationof the structure of a 120573-(1rarr 3)-D-glucan isolate from culturedfruit bodies of Sparassis crispardquoCarbohydrate Research vol 342no 17 pp 2611ndash2618 2007

[10] T Harada N Miura Y Adachi M Nakajima T Yadomae andN Ohno ldquoEffect of SCG 13-120573-D-glucan from Sparassis crispaon the hematopoietic response in cyclophosphamide inducedleukopenic micerdquo Biological and Pharmaceutical Bulletin vol25 no 7 pp 931ndash939 2002

[11] N Ohno T Harada S Masuzawa et al ldquoAntitumor activityand hematopoietic response of beta-glucan extracted froman edible and medicinal mushroom Sparassis crispa WulfFr(Aphyllophoromycetideae)rdquo International Journal of MedicinalMushrooms vol 4 pp 13ndash26 2002

[12] T Harada S Masuda M Arii et al ldquoSoy isoflavone aglyconemodulates A hematopoietic response in combination withsoluble 120573-glucan SCGrdquo Biological and Pharmaceutical Bulletinvol 28 no 12 pp 2342ndash2345 2005

[13] S Nameda T Harada N N Miura et al ldquoEnhancedcytokine synthesis of leukocytes by a 120573-glucan preparationSCG extracted from a medicinal mushroom Sparassis crispardquoImmunopharmacology and Immunotoxicology vol 25 no 3 pp321ndash335 2003

[14] T Harada N N Miura Y Adachi M Nakajima T Yadomaeand N Ohno ldquoIFN-120574 induction by SCG 13-120573-D-glucan fromSparassis crispa in DBA2 mice in vitrordquo Journal of Interferonand Cytokine Research vol 22 no 12 pp 1227ndash1239 2002

[15] T Harada N N Miura Y Adachi M Nakajima T Yadomaeand N Ohno ldquoGranulocyte-Macrophage Colony-StimulatingFactor (GM-CSF) regulates cytokine induction by 13-120573-D-glucan SCG in DBA2 mice in vitrordquo Journal of Interferon andCytokine Research vol 24 no 8 pp 478ndash489 2004

[16] T Harada N N Miura Y Adachi M Nakajima T Yadomaeand N Ohno ldquoAntibody to soluble 1316-120573-D-glucan SCGin sera of naive DBA2 micerdquo Biological and PharmaceuticalBulletin vol 26 no 8 pp 1225ndash1228 2003

[17] R TadaM Yoshikawa T Kuge et al ldquoGranulocytemacrophagecolony-stimulating factor is required for cytokine inductionby a highly 6-branched 13-120573-D-glucan from Aureobasidiumpullulans inmouse-derived splenocytesrdquo Immunopharmacologyand Immunotoxicology vol 33 no 2 pp 302ndash308 2011

[18] T Harada and N Ohno ldquoContribution of dectin-1 andgranulocyte macrophage-colony stimulating factor (GM-CSF)to immunomodulating actions of 120573-glucanrdquo InternationalImmunopharmacology vol 8 no 4 pp 556ndash566 2008

[19] M Sato H Sano D Iwaki et al ldquoDirect binding of toll-likereceptor 2 to zymosan and zymosan-induced NF-120581B activationand TNF-120572 secretion are down-regulated by lung collectinsurfactant protein Ardquo The Journal of Immunology vol 171 no1 pp 417ndash425 2003

[20] B P Thornton V Vetvicka M Pitman R C Goldman and GDRoss ldquoAnalysis of the sugar specificity andmolecular locationof the beta-glucan binding lectin site of complement receptortype 3 (CD11bCD18)rdquoThe Journal of Immunology vol 156 pp1235ndash1246 1996

[21] P J Rice J L Kelley G Kogan et al ldquoHuman monocytescavenger receptors are pattern recognition receptors for (1rarr

3)-120573-D-glucansrdquo Journal of Leukocyte Biology vol 72 no 1 pp140ndash146 2002

[22] J W Zimmerman J Lindermuth P A Fish G P PalaceT T Stevenson and D E DeMong ldquoA novel carbohydrate-glycosphingolipid interaction between a 120573-(1ndash3)-glucanimmunomodulator PGG-glucan and lactosylceramide ofhuman leukocytesrdquo The Journal of Biological Chemistry vol273 no 34 pp 22014ndash22020 1998

[23] G D Brown P R Taylor D M Reid et al ldquoDectin-1 is a major120573-glucan receptor on macrophagesrdquo Journal of ExperimentalMedicine vol 196 no 3 pp 407ndash412 2002

[24] D M Reid N A Gow and G D Brown ldquoPattern recognitionrecent insights fromDectin-1rdquoCurrent Opinion in Immunologyvol 21 no 1 pp 30ndash37 2009

[25] T Harada N N Miura Y Adachi M Nakajima T Yadomaeand N Ohno ldquoHighly expressed dectin-1 on bone marrow-derived dendritic cells regulates the sensitivity to 120573-glucan inDBA2 micerdquo Journal of Interferon and Cytokine Research vol28 no 8 pp 477ndash486 2007

[26] T Harada H Kawaminami N N Miura et al ldquoMechanism ofenhanced hematopoietic response by soluble 120573-glucan SCG incyclophosphamide-treated micerdquo Microbiology and Immunol-ogy vol 50 no 9 pp 687ndash700 2006

[27] A Shibata T H Hida K Ishibashi N N Miura Y Adachiand N Ohno ldquoDisruption of actin cytoskeleton enhancedcytokine synthesis of splenocytes stimulated with beta-glucanfrom the cauliflower medicinal mushroom Sparassis crispaWulfFr (higher Basidiomycetes) in vitrordquo International Journalof Medicinal Mushrooms vol 14 no 3 pp 257ndash269 2012

[28] H S Kim J Y Kim H S Ryu et al ldquoInduction of dendriticcell maturation by 120573-glucan isolated from Sparassis crispardquoInternational Immunopharmacology vol 10 no 10 pp 1284ndash1294 2010

[29] R Siepmann ldquoWachstumshemmung von Stammfaulepilzenund von Gremmeniella abietina durch Bacillus subtilisrdquo Euro-pean Journal of Forest Pathology vol 17 pp 59ndash64 1987

[30] S Woodward H Y Sultan D K Barrett and R B PearceldquoTwo new antifungal metabolites produced by Sparassis crispain culture and in decayed treesrdquo Journal of GeneralMicrobiologyvol 139 no 1 pp 153ndash159 1993

[31] H Kawagishi K Hayashi S Tokuyama N Hashimoto TKimura and M Dombo ldquoNovel bioactive compound fromthe Sparassis crispa mushroomrdquo Bioscience Biotechnology andBiochemistry vol 71 no 7 pp 1804ndash1806 2007

[32] S Kodani K Hayashi S Tokuyama et al ldquoOccurrenceand identification of chalcones from the culinary-medicinalcauliflower mushroom Sparassis crispa (Wulf) Fr (Aphyl-lophoromycetideae)rdquo International Journal of Medicinal Mush-rooms vol 10 no 4 pp 331ndash336 2008

[33] S Kodani K Hayashi M Hashimoto T Kimura M Domboand H Kawagishi ldquoNew sesquiterpenoid from the mushroomSparassis crispardquo Bioscience Biotechnology and Biochemistryvol 73 no 1 pp 228ndash229 2009

[34] K Yoshikawa N Kokudo T Hashimoto K Yamamoto TInose and T Kimura ldquoNovel phthalide compounds fromSparassis crispa (Hanabiratake) Hanabiratakelide A-C exhibit-ing anti-cancer related activityrdquo Biological and PharmaceuticalBulletin vol 33 no 8 pp 1355ndash1359 2010

[35] JWangHXWang andT BNg ldquoApeptidewithHIV-1 reversetranscriptase inhibitory activity from the medicinal mushroomRussula paludosardquo Peptides vol 28 no 3 pp 560ndash565 2007


[36] H Yoshitomi E Iwaoka M Kubo M Shibata and M GaoldquoBeneficial effect of Sparassis crispa on stroke through activationof AkteNOS pathway in brain of SHRSPrdquo Journal of NaturalMedicines vol 65 no 1 pp 135ndash141 2011

[37] S Y Lee Y G Lee S E Byeon et al ldquoMitogen activated proteinkinases are prime signalling enzymes in nitric oxide productioninduced by soluble 120573-glucan from Sparassis crispardquo Archives ofPharmacal Research vol 33 no 11 pp 1753ndash1760 2010

[38] K Yamamoto and T Kimura ldquoOrally and topically admin-istered Sparassis crispa (Hanabiratake) improves healing ofskin wounds in mice with streptozotocin-induced diabetes rdquoBioscience Biotechnology and Biochemistry In press

[39] K Yamamoto and T Kimura ldquoDietary Sparassis crispa (Han-abiratake) ameliorates plasma levels of adiponectin and glucosein type 2 diabetic micerdquo Journal of Health Science vol 56 no 5pp 541ndash546 2010

[40] S Wu T F Lu S Wu and W J Guan ldquoResearch progress in Gprotein-coupled receptor 40rdquoYi Chuan vol 35 no 4 pp 27ndash342013

[41] K Yoshikawa T Hashimoto A Hirasawa et al ldquoUnsaturatedfatty acids derived from Sparassis crispa fruiting body andormycelium for inhibiting diabetes mellitusrdquo Japanese KokaiTokkyo Koho P2010-59106A (March 2010)

[42] A H Kwon Z Qiu M Hashimoto K Yamamoto and TKimura ldquoEffects of medicinal mushroom (Sparassis crispa)on wound healing in streptozotocin-induced diabetic ratsrdquoAmerican Journal of Surgery vol 197 no 4 pp 503ndash509 2009

[43] K Yamamoto Y Nishikawa T Kimura M Dombo N Mat-suura and A Sugitachi ldquoAntitumor activities of low molecularweight fraction derived from the cultured fruit body of Sparas-sis crispa in tumor-bearing micerdquo Nippon Shokuhin KagakuKogaku Kaishi vol 54 no 9 pp 419ndash423 2007

[44] M Yao K Yamamoto T Kimura and M Dombo ldquoEffectsof Hanabiratake (Sparassis crispa) on allergic rhinitis in OVA-sensitized micerdquo Food Science and Technology Research vol 14no 6 pp 589ndash594 2008

[45] H H Kim S Lee T S Singh J K Choi T Y Shin andS H Kim ldquoSparassis crispa suppresses mast cell-mediatedallergic inflammation role of calcium mitogen-activated pro-tein kinase and nuclear factor-120581Brdquo International Journal ofMolecular Medicine vol 30 no 2 pp 344ndash350 2012

[46] T Kimura ldquoThe physiological function of beta-glucan inSparassis crispardquo Food Processing and Ingredients vol 44 no 11pp 14ndash15 2009

[47] N Ohno S Nameda T Harada et al ldquoImmunomodulatingactivity of a beta-glucan preparation SCG extracted froma culinary-medicinal mushroom Sparassis crispa WulfFr(Aphyllophoromycetideae) and application to cancer patientsrdquoInternational Journal of Medicinal Mushrooms vol 5 pp 359ndash368 2003

[48] J Chen W Gu and K Zhao ldquoThe role of PI3KAkt pathwayin 120573-glucan-induced dendritic cell maturationrdquo InternationalImmunopharmacology vol 11 no 4 p 529 2011

[49] HG Park Y Y Shim S O Choi andWM Park ldquoNewmethoddevelopment for nanoparticle extraction of water-soluble 120573-(1rarr 3)-D-glucan from edible mushrooms Sparassis crispa andPhellinus linteusrdquo Journal of Agricultural and Food Chemistryvol 57 no 6 pp 2147ndash2154 2009

[50] T Uchiyama ldquoSparassis crispa extractions resveratrol andcollagen peptide as antiaging agents and food supplementsrdquoJapanese Tokkyo Koho P5016535 (June 2012)

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Anatomy Research International

PeptidesInternational Journal of


Hindawi Publishing Corporation httpwwwhindawicom

International Journal of

Volume 2014

Zoology


Molecular Biology International

GenomicsInternational Journal of


The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014


BioinformaticsAdvances in

Marine BiologyJournal of



Signal TransductionJournal of


BioMed Research International

Evolutionary BiologyInternational Journal of



Biochemistry Research International

ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Genetics Research International


Advances in

Virolog y

Hindawi Publishing Corporationhttpwwwhindawicom

Nucleic AcidsJournal of

Volume 2014

Stem CellsInternational



Enzyme Research



Microbiology


10 cm

Figure 1 Sparassis crispaWulfFr [2]

Table 1 Approximate composition of Sparassis crispa (per 100 g drysample)

Components Amount (g)Protein 134Fat 20Ash 18Carbohydrate 215Dietary fiber (DF) 612Beta-glucan from DF 435

21 Polysaccharide (Beta-Glucan)

211 Primary Structure Using chemical enzymatic andNMR analyses it was shown that the primary structureof a purified beta-glucan (designated SCG) obtained fromcultured fruiting bodies of S crispa is a 6-branched 13-beta-glucan with one branch in approximately every 3 main chainunits (Figure 2) [7ndash9]

212 Biological Activities Tumor size in cancerous (Sarcoma180) ICR mice was dose-dependently decreased after 5 weeksof oral administration of S crispa (10 or 100mgkg) incomparison to a control group Furthermore the survival rateof these model mice was higher when similarly treated withS crispa [2] Since SCG content in dry powder of S crispawasmeasured to bemore than 40 SCGwas likely be responsiblefor this antitumor effect

Ohno et al prepared polysaccharide fractions from thefruiting bodies of cultured S crispa and showed their anti-tumor activity against the solid form of Sarcoma 180 in ICRmice with strong vascular dilation and hemorrhage reactions[7] Furthermore intraperitoneal and oral SCG over a widerange of concentrations enhanced hematopoietic responsesin mice with leukopenia induced by cyclophosphamide (CYa DNA-alkylating agent) [10 11] This effect was augmentedin combination with isoflavone aglycone [12] SCG was also

shown to stimulate leukocytes to produce cytokines such asIL-8 in whole-cell cultures of human peripheral blood [13]and in mouse splenocytes [14]

Yamamoto et al reported antiangiogenic and anti-metastatic effects of SCG on neoplasm by using differentanimal models [8] Oral administration of SCG suppressedB16-F10 cell-induced angiogenesis in a dorsal air sac assayusing ICR mice and suppressed vascular endothelial growthfactor induced neovascularization in a Matrigel plug assayusing C57BL6J mice Furthermore it suppressed the growthand number of metastatic tumor foci in the lung alongwith primary tumor growth in a C57BL6J mice model ofspontaneous metastasis From these findings it is apparentthat the oral administration of SCG exerts a suppressive effecton tumor growth and metastasis in the lung through theinhibition of tumor-induced angiogenesis

Taken together these results demonstrate that SCG ex-hibits various biological activities including antitumoreffects enhancement of the hematopoietic response andinduction of cytokine production in vivo and in vitro

213 Mechanisms Harada et al reported strain-specificdifferences of the reactivity of mice to SCG with DBA1 andDBA2 mice being highly sensitive to SCG In splenocytesderived from various inbred strains of mice interferon-120574(IFN-120574) production was not induced by SCG Howeversplenocytes from naıve DBA1 and DBA2 mice stronglyreact with SCG to produce IFN-120574 [14] Furthermorein addition to IFN-120574 cytokines induced by SCG werescreened for and found to include tumor necrosis factor-120572(TNF-120572) granulocyte-macrophage colony-stimulating factor(GM-CSF) and interleukin-12 (IL-12p70) [15] Since the seraof naıve DBA1 and DBA2 mice contain significantly highertiters of antibody against SCG than other strains of mice [16]it seems likely that these mice strains are sensitive to SCGThus DBA1 and DBA2 mice would be useful models forfuture studies of SCG

Harada et al further demonstrated that GM-CSF was oneof the key factors in reactivity to SCG in DBA2 mice [15 17]Neutralizing GM-CSF using an anti-GM-CSF monoclonalantibody significantly inhibited IFN-120574 TNF-120572 and IL-12p70elicited by SCG The splenocytes in various strains of miceshowed similar patterns of cytokine production in responseto SCG cotreatment in the presence of recombinant murineGM-CSF The high sensitivity to SGG shown by DBA1and DBA2 mice may be attributable to differences of theirregulation of GM-CSF compared with that in other mice

Harada and Ohno also proposed an interesting model forthe mechanism of cytokine induction by SCG in DBA2mice[18] Broadly speaking SCG directly induces adherent cellsto produce TNF-120572 and IL-12p70 whereas cell-cell contactmediated by the association of CD4+ T cells expressing LFA-1and antigen-presenting cells such as dendritic cells expressingICAM-1 is required for the induction of IFN-120574 and GM-CSFby SCG

Neutrophils macrophages and dendritic cells expressseveral receptors capable of recognizing beta-glucan in its


O

H

HO

H

O

H

H

OHH

OH

O

H

H

H

HOHH

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H

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HO

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OH

H

O

OH

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119899

O




























N

OH

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OH

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OHOH

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HO OH

OH O

OH

OH O

CH3O

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OH

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CH3O CH3

OCH3 OCH3

CH3

OCH3

CH3OCH3O

(1) (2)

(3) (4)

(5)

(6) (7)

(8) (9) (10)


























References





























































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


O

H

HO

H

O

H

H

OHH

OH

O

H

H

H

HOHH

O

H

HO

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H

H

OHHO

OH

O

O

HHO

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HO

H

H

OH

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O

OH

OH

119899

O




























N

OH

OO

O

N

OH

OO

O

OHOH

H

HO OH

OH O

OH

OH O

CH3O

O

OH

HO

O

O

HO

OOH

O

HO

HOO

OH

OH O

HO

OH O

CH3O CH3

OCH3 OCH3

CH3

OCH3

CH3OCH3O

(1) (2)

(3) (4)

(5)

(6) (7)

(8) (9) (10)


























References





























































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology




















N

OH

OO

O

N

OH

OO

O

OHOH

H

HO OH

OH O

OH

OH O

CH3O

O

OH

HO

O

O

HO

OOH

O

HO

HOO

OH

OH O

HO

OH O

CH3O CH3

OCH3 OCH3

CH3

OCH3

CH3OCH3O

(1) (2)

(3) (4)

(5)

(6) (7)

(8) (9) (10)


























References





























































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology























References





























































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology






















































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

review article natural products and biological...

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