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Natural Product Research: Formerly Natural Product Letters Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/gnpl20
Bioactive metabolites isolated from Penicillium sp. YY-20, the endophytic fungus from Ginkgo biloba a
a
a
a
a
Yuan Yuan , Jun-Mian Tian , Jian Xiao , Qi Shao & Jin-Ming Gao a
College of Science, Northwest A&F University, YanglingShaanxi712100, P.R. China Published online: 21 Oct 2013.
To cite this article: Yuan Yuan, Jun-Mian Tian, Jian Xiao, Qi Shao & Jin-Ming Gao , Natural Product Research (2013): Bioactive metabolites isolated from Penicillium sp. YY-20, the endophytic fungus from Ginkgo biloba, Natural Product Research: Formerly Natural Product Letters, DOI: 10.1080/14786419.2013.850686 To link to this article: http://dx.doi.org/10.1080/14786419.2013.850686
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content. This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://www.tandfonline.com/page/termsand-conditions
Natural Product Research, 2013 http://dx.doi.org/10.1080/14786419.2013.850686
SHORT COMMUNICATION Bioactive metabolites isolated from Penicillium sp. YY-20, the endophytic fungus from Ginkgo biloba Yuan Yuan, Jun-Mian Tian, Jian Xiao, Qi Shao and Jin-Ming Gao* College of Science, Northwest A&F University, Yangling, Shaanxi 712100, P.R. China
Downloaded by [Moskow State Univ Bibliote] at 17:37 06 February 2014
(Received 4 June 2013; final version received 5 September 2013) Six known metabolites, adenosine (1), methyl b-D -ribofuranoside (2), adenine (3), 20 -deoxyadenosine (4), 3-methylpiperazine-2,5-dione (5) and 20 -deoxyuridine (6), were isolated from the extracts of the endophytic fungus Penicillium sp. YY-20 isolated from the root of Ginkgo biloba, and their structures were elucidated by spectroscopic methods. The antioxidant and growth-promoting activities of these compounds were first evaluated. The results indicated that compounds 1, 3 and 4 exhibited potential DPPH-scavenging activities compared with positive control. In addition, all the compounds (except 5) stimulated seed germination of Raphanus sativus, Brassica napus and Brassica chinensis but had weak stimulating effect on their root and hypocotyl growth. Keywords: Penicillium sp; endophytic fungus; nucleoside; antioxidant activity; plant growth-promoting activity
1. Introduction Endophytic fungi have been recognised as an important source of a variety of structurally novel active secondary metabolites with anti-cancer, anti-microbial and other biological activities (Li et al. 2011). In our continuous screening for bioactive metabolites from plant endophytes (Gao et al. 2013), we investigated the metabolites of the extract of the endophytic fungus Penicillium sp. YY-20 residing in the root of Ginkgo biloba, leading to the isolation of six compounds 1– 6. The main aim of this study was to isolate metabolites with potential bioactivity from the endophytic fungus of G. biloba. In this study, we report the isolation, antioxidant and growth-promoting activities of these products from this endophyte. 2. Results and discussion The EtOAc-soluble material from the methanol extract of the endophytic fungus was subjected to repeated silica gel column chromatography (CC), followed by purification on Sephadex LH-20, reversed phase CC and preparative TLC to afford six compounds 1– 6 (Figure 1). These compounds were identified as adenosine (Chenon et al. 1975), methyl b-D -ribofuranoside (Chapeau & Marnett 1993; Church et al. 1997), adenine (Gao et al. 2001), 20 -deoxyadenosine (Chenon et al. 1975), 3-methylpiperazine-2,5-dione (Nakatsuka et al. 1980) and 20 -deoxyuridine (Nie et al. 1994) by analysis of MS and NMR data as well as by comparison with literature data. All compounds 1– 6 were examined for DPPH-scavenging activities using the method reported (Zheng et al. 2010). Of the tested compounds, 1 had higher DPPH-scavenging activity, with an IC50 value of 2.87 mg/mL, than the positive controls TBHQ (IC50 4.57 mg/mL) and VC
*Corresponding author. Email: [email protected] q 2013 Taylor & Francis
2
Y. Yuan et al. NH2 N N
HO
NH2
N N
HO
OCH3
O
N
N
O
N H
N
OH OH OH OH 1
2
3
Downloaded by [Moskow State Univ Bibliote] at 17:37 06 February 2014
NH2 N N
HO
O
O
N
NH NH
N
O
HO
HN
N
O
O
O
OH
OH 4
5
6
Figure 1. Structures of compounds 1 – 6.
(IC50 8.61 mg/mL), while 3 and 4 with IC50 values of 8.46 and 8.37 mg/mL, respectively, are similar to that of VC. In contrast, the other compounds showed weak ability (IC50 . 100 mg/ mL). The order of DPPH radical-scavenging abilities of these compounds was 1 . 4 . 3 . 6 . 2 . 5 (Table 1). So, the formation of stable radical intermediate products with a large conjugated system derived from 1, 3 and 4 is essential to the antioxidant activity. On the other hand, nucleoside adenosine (1) has been reported to significantly stimulate the edible mushroom production (Domondon et al. 2004). So, the allelopathic activity of these metabolites was tested in this work by the method reported by our group (Li et al. 2011). Most of them (except 5) exhibited stimulatory effects on the seed germination of three plants, Raphanus sativus, Brassica napus and Brassica chinensis, with seed germination rates of 20 –70%, compared with the positive control, glyphosate (Figure 2). Notably, compounds 3 and 4 markedly promoted the seed germination of B. chinensis in a concentration-dependent manner (12.5 – 400 mg/mL), showing EC50 values of 78.73 and 54.69 mg/mL, respectively. Moreover, almost all compounds showed weak stimulating effect on the root and hypocotyl growth of these acceptor plants (100 –400 mg/mL). This growth-promoting activity may be ascribed to the presence of adenine and its nucleosides (Domondon et al. 2004). Compounds 1, 3 and 4 have a large conjugated system and many hydroxyls, so they show the strong anti-oxidant activity. However, the growth-promoting activity of compounds 3 and 4 may be ascribed to the growth regulation of the nucleosides, which are most important Table 1. The antioxidant activity of six compounds 1 – 6. Compounds 1 2 3 4
IC50 (mg/mL)
Compounds
IC50 (mg/mL)
2.87 ^ 0.02 .100 8.46 ^ 0.03 8.37 ^ 0.04
5 6 TBHQ VC
.100 .100 4.57 ^ 0.02 8.61 ^ 0.02
Downloaded by [Moskow State Univ Bibliote] at 17:37 06 February 2014
Natural Product Research
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Figure 2. The percentage of relative germination of three kinds of seeds. Note: % seed germination , 0, inhibiting effect; . 0, stimulating effect; CK, glyphosate; R. s., Raphanus sativus; B. n., Brassica napus; B. c., Brassica chinensis.
growth-promoting substances for life growth. As a result, there is no relation between the antioxidant activity of compound and seed germination. This study reported the isolation and identification of six metabolites from Penicillium sp. YY20. Among them, adenine derivatives 1, 3 and 4 exerted strong antioxidant activity, and 3 and 4 exhibited a strong stimulating effect on the seed germination of B. chinensis. These data indicate that 1, 3 and 4 were first identified as the major bioactive substances in this endophytic fungus. Supplementary material Experimental details relating to this article are available online. Acknowledgements This work is financially supported by the earmarked fund for National Key Technology R&D Program of the Ministry of Science and Technology of China (2012BAD36B04).
References Chapeau MC, Marnett LJ. 1993. Pentenyl ribosides: new reagents for purine nucleoside synthesis. J Org Chem. 58:7258–7262. Chenon MT, Pugmire RJ, Grant DM, Panzica RP, Townsend LB. 1975. Carbon-13 magnetic resonance. XXV. Basic set of parameters for the investigation of tautomerism in purines established from carbon-13 magnetic resonance studies using certain purines and pyrrolo [2, 3-d] pyrimidines. J Am Chem Soc. 97:4627–4636. Church TJ, Carmichael I, Serianni AS. 1997. 13C-1H and 13C-13C spin-coupling constants in methyl b-d-ribofuranoside and methyl 2-deoxy-b-d-erythro-pentofuranoside: correlations with molecular structure and conformation. J Am Chem Soc. 119:8946– 8964. Domondon DL, He WD, De Kimpe N, Ho¨fte M, Poppe J. 2004. b-Adenosine, a bioactive compound in grass chaff stimulating mushroom production. Phytochemistry. 65:181–187.
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Y. Yuan et al.
Downloaded by [Moskow State Univ Bibliote] at 17:37 06 February 2014
Gao JM, Dong ZL, Liu JK. 2001. A new ceramide from basidiomycetes Russula cyanoxantha. Lipids. 36(2):175–180. Gao JM, Yang SX, Qin JC. 2013. Azaphilones: chemistry and biology. Chem Rev. 113(7):4755–4811. Li HQ, Li XJ, Wang YL, Zhang Q, Zhang AL, Gao JM, Zhang XC. 2011. Antifungal metabolites from Chaetomium globosum, an endophytic fungus in Ginkgo biloba. Biochem Syst Ecol. 39:876–879. Nakatsuka S, Miyazaki H, Goto T. 1980. Total synthesis of (^)-neoechinulin A, an indole alkaloid containing oxidized diketopiperazine. Tetrahedron Lett. 21:2817–2820. Nie SQ, Majarais I, Kwan CY, Epand RM. 1994. Analogues of tetramethylpyrazine affect membrane fluidity of liposomes: relationship to their biological activities. Eur J Pharmacol Mol Pharmacol. 266:11–18. Zheng CD, Li G, Li HQ, Xu XJ, Gao JM, Zhang AL. 2010. DPPH-scavenging activities and structure-activity relationships of phenolic compounds. Nat Prod Commun. 5:1759–1765.
Natural Product Research: Formerly Natural Product Letters Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/gnpl20
Bioactive metabolites isolated from Penicillium sp. YY-20, the endophytic fungus from Ginkgo biloba a
a
a
a
a
Yuan Yuan , Jun-Mian Tian , Jian Xiao , Qi Shao & Jin-Ming Gao a
College of Science, Northwest A&F University, YanglingShaanxi712100, P.R. China Published online: 21 Oct 2013.
To cite this article: Yuan Yuan, Jun-Mian Tian, Jian Xiao, Qi Shao & Jin-Ming Gao , Natural Product Research (2013): Bioactive metabolites isolated from Penicillium sp. YY-20, the endophytic fungus from Ginkgo biloba, Natural Product Research: Formerly Natural Product Letters, DOI: 10.1080/14786419.2013.850686 To link to this article: http://dx.doi.org/10.1080/14786419.2013.850686
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content. This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://www.tandfonline.com/page/termsand-conditions
Natural Product Research, 2013 http://dx.doi.org/10.1080/14786419.2013.850686
SHORT COMMUNICATION Bioactive metabolites isolated from Penicillium sp. YY-20, the endophytic fungus from Ginkgo biloba Yuan Yuan, Jun-Mian Tian, Jian Xiao, Qi Shao and Jin-Ming Gao* College of Science, Northwest A&F University, Yangling, Shaanxi 712100, P.R. China
Downloaded by [Moskow State Univ Bibliote] at 17:37 06 February 2014
(Received 4 June 2013; final version received 5 September 2013) Six known metabolites, adenosine (1), methyl b-D -ribofuranoside (2), adenine (3), 20 -deoxyadenosine (4), 3-methylpiperazine-2,5-dione (5) and 20 -deoxyuridine (6), were isolated from the extracts of the endophytic fungus Penicillium sp. YY-20 isolated from the root of Ginkgo biloba, and their structures were elucidated by spectroscopic methods. The antioxidant and growth-promoting activities of these compounds were first evaluated. The results indicated that compounds 1, 3 and 4 exhibited potential DPPH-scavenging activities compared with positive control. In addition, all the compounds (except 5) stimulated seed germination of Raphanus sativus, Brassica napus and Brassica chinensis but had weak stimulating effect on their root and hypocotyl growth. Keywords: Penicillium sp; endophytic fungus; nucleoside; antioxidant activity; plant growth-promoting activity
1. Introduction Endophytic fungi have been recognised as an important source of a variety of structurally novel active secondary metabolites with anti-cancer, anti-microbial and other biological activities (Li et al. 2011). In our continuous screening for bioactive metabolites from plant endophytes (Gao et al. 2013), we investigated the metabolites of the extract of the endophytic fungus Penicillium sp. YY-20 residing in the root of Ginkgo biloba, leading to the isolation of six compounds 1– 6. The main aim of this study was to isolate metabolites with potential bioactivity from the endophytic fungus of G. biloba. In this study, we report the isolation, antioxidant and growth-promoting activities of these products from this endophyte. 2. Results and discussion The EtOAc-soluble material from the methanol extract of the endophytic fungus was subjected to repeated silica gel column chromatography (CC), followed by purification on Sephadex LH-20, reversed phase CC and preparative TLC to afford six compounds 1– 6 (Figure 1). These compounds were identified as adenosine (Chenon et al. 1975), methyl b-D -ribofuranoside (Chapeau & Marnett 1993; Church et al. 1997), adenine (Gao et al. 2001), 20 -deoxyadenosine (Chenon et al. 1975), 3-methylpiperazine-2,5-dione (Nakatsuka et al. 1980) and 20 -deoxyuridine (Nie et al. 1994) by analysis of MS and NMR data as well as by comparison with literature data. All compounds 1– 6 were examined for DPPH-scavenging activities using the method reported (Zheng et al. 2010). Of the tested compounds, 1 had higher DPPH-scavenging activity, with an IC50 value of 2.87 mg/mL, than the positive controls TBHQ (IC50 4.57 mg/mL) and VC
*Corresponding author. Email: [email protected] q 2013 Taylor & Francis
2
Y. Yuan et al. NH2 N N
HO
NH2
N N
HO
OCH3
O
N
N
O
N H
N
OH OH OH OH 1
2
3
Downloaded by [Moskow State Univ Bibliote] at 17:37 06 February 2014
NH2 N N
HO
O
O
N
NH NH
N
O
HO
HN
N
O
O
O
OH
OH 4
5
6
Figure 1. Structures of compounds 1 – 6.
(IC50 8.61 mg/mL), while 3 and 4 with IC50 values of 8.46 and 8.37 mg/mL, respectively, are similar to that of VC. In contrast, the other compounds showed weak ability (IC50 . 100 mg/ mL). The order of DPPH radical-scavenging abilities of these compounds was 1 . 4 . 3 . 6 . 2 . 5 (Table 1). So, the formation of stable radical intermediate products with a large conjugated system derived from 1, 3 and 4 is essential to the antioxidant activity. On the other hand, nucleoside adenosine (1) has been reported to significantly stimulate the edible mushroom production (Domondon et al. 2004). So, the allelopathic activity of these metabolites was tested in this work by the method reported by our group (Li et al. 2011). Most of them (except 5) exhibited stimulatory effects on the seed germination of three plants, Raphanus sativus, Brassica napus and Brassica chinensis, with seed germination rates of 20 –70%, compared with the positive control, glyphosate (Figure 2). Notably, compounds 3 and 4 markedly promoted the seed germination of B. chinensis in a concentration-dependent manner (12.5 – 400 mg/mL), showing EC50 values of 78.73 and 54.69 mg/mL, respectively. Moreover, almost all compounds showed weak stimulating effect on the root and hypocotyl growth of these acceptor plants (100 –400 mg/mL). This growth-promoting activity may be ascribed to the presence of adenine and its nucleosides (Domondon et al. 2004). Compounds 1, 3 and 4 have a large conjugated system and many hydroxyls, so they show the strong anti-oxidant activity. However, the growth-promoting activity of compounds 3 and 4 may be ascribed to the growth regulation of the nucleosides, which are most important Table 1. The antioxidant activity of six compounds 1 – 6. Compounds 1 2 3 4
IC50 (mg/mL)
Compounds
IC50 (mg/mL)
2.87 ^ 0.02 .100 8.46 ^ 0.03 8.37 ^ 0.04
5 6 TBHQ VC
.100 .100 4.57 ^ 0.02 8.61 ^ 0.02
Downloaded by [Moskow State Univ Bibliote] at 17:37 06 February 2014
Natural Product Research
3
Figure 2. The percentage of relative germination of three kinds of seeds. Note: % seed germination , 0, inhibiting effect; . 0, stimulating effect; CK, glyphosate; R. s., Raphanus sativus; B. n., Brassica napus; B. c., Brassica chinensis.
growth-promoting substances for life growth. As a result, there is no relation between the antioxidant activity of compound and seed germination. This study reported the isolation and identification of six metabolites from Penicillium sp. YY20. Among them, adenine derivatives 1, 3 and 4 exerted strong antioxidant activity, and 3 and 4 exhibited a strong stimulating effect on the seed germination of B. chinensis. These data indicate that 1, 3 and 4 were first identified as the major bioactive substances in this endophytic fungus. Supplementary material Experimental details relating to this article are available online. Acknowledgements This work is financially supported by the earmarked fund for National Key Technology R&D Program of the Ministry of Science and Technology of China (2012BAD36B04).
References Chapeau MC, Marnett LJ. 1993. Pentenyl ribosides: new reagents for purine nucleoside synthesis. J Org Chem. 58:7258–7262. Chenon MT, Pugmire RJ, Grant DM, Panzica RP, Townsend LB. 1975. Carbon-13 magnetic resonance. XXV. Basic set of parameters for the investigation of tautomerism in purines established from carbon-13 magnetic resonance studies using certain purines and pyrrolo [2, 3-d] pyrimidines. J Am Chem Soc. 97:4627–4636. Church TJ, Carmichael I, Serianni AS. 1997. 13C-1H and 13C-13C spin-coupling constants in methyl b-d-ribofuranoside and methyl 2-deoxy-b-d-erythro-pentofuranoside: correlations with molecular structure and conformation. J Am Chem Soc. 119:8946– 8964. Domondon DL, He WD, De Kimpe N, Ho¨fte M, Poppe J. 2004. b-Adenosine, a bioactive compound in grass chaff stimulating mushroom production. Phytochemistry. 65:181–187.
4
Y. Yuan et al.
Downloaded by [Moskow State Univ Bibliote] at 17:37 06 February 2014
Gao JM, Dong ZL, Liu JK. 2001. A new ceramide from basidiomycetes Russula cyanoxantha. Lipids. 36(2):175–180. Gao JM, Yang SX, Qin JC. 2013. Azaphilones: chemistry and biology. Chem Rev. 113(7):4755–4811. Li HQ, Li XJ, Wang YL, Zhang Q, Zhang AL, Gao JM, Zhang XC. 2011. Antifungal metabolites from Chaetomium globosum, an endophytic fungus in Ginkgo biloba. Biochem Syst Ecol. 39:876–879. Nakatsuka S, Miyazaki H, Goto T. 1980. Total synthesis of (^)-neoechinulin A, an indole alkaloid containing oxidized diketopiperazine. Tetrahedron Lett. 21:2817–2820. Nie SQ, Majarais I, Kwan CY, Epand RM. 1994. Analogues of tetramethylpyrazine affect membrane fluidity of liposomes: relationship to their biological activities. Eur J Pharmacol Mol Pharmacol. 266:11–18. Zheng CD, Li G, Li HQ, Xu XJ, Gao JM, Zhang AL. 2010. DPPH-scavenging activities and structure-activity relationships of phenolic compounds. Nat Prod Commun. 5:1759–1765.
