Full Text:   <2691>

Summary:  <1900>

CLC number: Q815

On-line Access: 2015-12-04

Received: 2015-06-17

Revision Accepted: 2015-08-04

Crosschecked: 2015-11-11

Cited: 4

Clicked: 4580

Citations:  Bibtex RefMan EndNote GB/T7714

 ORCID:

Guo-qing He

http://orcid.org/0000-0002-1177-8016

-   Go to

Article info.
Open peer comments

Journal of Zhejiang University SCIENCE B 2015 Vol.16 No.12 P.998-1010

http://doi.org/10.1631/jzus.B1500147


Submerged fermentation production and characterization of intracellular triterpenoids from Ganoderma lucidum using HPLC-ESI-MS


Author(s):  Mei-lin Cui, Huan-yi Yang, Guo-qing He

Affiliation(s):  Zhejiang Provincial Key Laboratory of Food Microbiology, College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou 310058, China

Corresponding email(s):   gqhe@zju.edu.cn

Key Words:  Ganoderma lucidum, Wort, Response surface methodology, Ganoderic acids, HPLC-ESI-MS


Mei-lin Cui, Huan-yi Yang, Guo-qing He. Submerged fermentation production and characterization of intracellular triterpenoids from Ganoderma lucidum using HPLC-ESI-MS[J]. Journal of Zhejiang University Science B, 2015, 16(12): 998-1010.

@article{title="Submerged fermentation production and characterization of intracellular triterpenoids from Ganoderma lucidum using HPLC-ESI-MS",
author="Mei-lin Cui, Huan-yi Yang, Guo-qing He",
journal="Journal of Zhejiang University Science B",
volume="16",
number="12",
pages="998-1010",
year="2015",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1500147"
}

%0 Journal Article
%T Submerged fermentation production and characterization of intracellular triterpenoids from Ganoderma lucidum using HPLC-ESI-MS
%A Mei-lin Cui
%A Huan-yi Yang
%A Guo-qing He
%J Journal of Zhejiang University SCIENCE B
%V 16
%N 12
%P 998-1010
%@ 1673-1581
%D 2015
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1500147

TY - JOUR
T1 - Submerged fermentation production and characterization of intracellular triterpenoids from Ganoderma lucidum using HPLC-ESI-MS
A1 - Mei-lin Cui
A1 - Huan-yi Yang
A1 - Guo-qing He
J0 - Journal of Zhejiang University Science B
VL - 16
IS - 12
SP - 998
EP - 1010
%@ 1673-1581
Y1 - 2015
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1500147


Abstract: 
As the main bioactive metabolites of Ganoderma lucidum, triterpenoids have various pharmacological effects. In this paper, the nutritional requirements and culture conditions of a submerged culture of G. lucidum were optimized using the response surface methodology; maximum mycelia biomass and intracellular triterpenoid production reached 1.87 g/100 ml and 93.21 mg/100 ml, respectively, for a culture consisting of wort 4.10% (0.041 g/ml) and yeast extract 1.89% (0.0189 g/ml), pH 5.40. For the first time, we established that wort, which is cheap and abundant, can replace the more commonly used glucose as the sole source of carbohydrate. Using high-performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS), 10 major ganoderic acids were tentatively identified based on the predominant fragmentation pathways with the elimination of H2O and CO2, as well as cleavage of the D-ring.

利用液体发酵提高灵芝三萜产量及HPLC-ESI-MS鉴定灵芝菌丝中灵芝酸成分

目的:通过优化液体发酵培养基提高灵芝三萜产量,并对其三萜成分进行初步鉴定。
创新点:麦芽作为啤酒生产原料,含有丰富的糖类及其他营养物质,并且价格低廉,来源广泛。本文首次将麦芽汁作为碳源物质以提高灵芝三萜产量,并对其所含灵芝酸成分进行初步鉴定。
方法:以灵芝菌丝生物量和胞内三萜产量为检测指标,对其液体发酵培养基中的碳(C)源和氮(N)源进行筛选,并利用单因素实验确定其相应浓度,再经响应面法优化液体发酵培养基(C源、N源、pH)(表1)。另一方面,酸碱转化法纯化灵芝三萜提取物后,利用高效液相色谱-电喷雾-质谱(HPLC-ESI-MS)法初步鉴定灵芝所含灵芝酸成分(表2)。
结论:本实验响应面结果显示:当培养基含麦芽汁4.1%,酵母浸出粉1.89%,pH为5.40时,灵芝液体发酵菌丝生物量及胞内三萜产量均达到最优,分别为1.87 g/100 ml和93.21 mg/100 ml。经HPLC-ESI-MS鉴定,初步确定其含有10种灵芝酸成分。

关键词:灵芝;麦芽汁;响应面;灵芝酸;高效液相色谱-电喷雾-质谱(HPLC-ESI-MS)

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

Reference

[1]Cheng, C.R., Yang, M., Wu, Z.Y., et al., 2011. Fragmentation pathways of oxygenated tetracyclic triterpenoids and their application in the qualitative analysis of Ganoderma lucidum by multistage tandem mass spectrometry. Rapid Commun. Mass Spectrom., 25(9):1323-1335.

[2]Ding, N., Yang, Q., Huang, S.S., et al., 2010. Separation and determination of four ganoderic acids from dried fermentation mycelia powder of Ganoderma lucidum by capillary zone electrophoresis. J. Pharm. Biomed. Anal., 53(5):1224-1230.

[3]Fang, Q.H., Zhong, J.J., 2002a. Effect of initial pH on production of ganoderic acid and polysaccharide by submerged fermentation of Ganoderma lucidum. Process Biochem., 37(7):769-774.

[4]Fang, Q.H., Zhong, J.J., 2002b. Submerged fermentation of higher fungus Ganoderma lucidum for production of valuable bioactive metabolites-ganoderic acid and polysaccharide. Biochem. Eng. J., 10(1):61-65.

[5]Fang, Q.H., Tang, Y.J., Zhong, J.J., 2002. Significance of inoculation density control in production of polysaccharide and ganoderic acid by submerged culture of Ganoderma lucidum. Process Biochem., 37(12):1375-1379.

[6]Guo, X.Y., Han, J., Ye, M., et al., 2012. Identification of major compounds in rat bile after oral administration of total triterpenoids of Ganoderma lucidum by high-performance liquid chromatography with electrospray ionization tandem mass spectrometry. J. Pharm. Biomed. Anal., 63:29-39.

[7]Guo, X.Y., Liu, D., Ye, M., et al., 2013. Structural characterization of minor metabolites and pharmacokinetics of ganoderic acid C2 in rat plasma by HPLC coupled with electrospray ionization tandem mass spectrometry. J. Pharm. Biomed. Anal., 75:64-73.

[8]Keypour, S., Rafati, H., Riahi, H., et al., 2010. Qualitative analysis of ganoderic acids in Ganoderma lucidum from Iran and China by RP-HPLC and electrospray ionisation-mass spectrometry (ESI-MS). Food Chem., 119(4):1704-1708.

[9]Kim, H.H., Na, J.G., Chang, Y.K., et al., 2004. Optimization of submerged culture conditions for mycelial growth and exopolysaccharides production by Agaricus blazei. J. Microbiol. Biotechnol., 14(5):944-951.

[10]Kim, H.M., Paik, S.Y., Ra, K.S., et al., 2006. Enhanced production of exopolysaccharides by fed-batch culture of Ganoderma resinaceum DG-6556. J. Microbiol., 44(2):233-242.

[11]Lin, J.H., Yang, S.S., 2006. Mycelium and polysaccharide production of Agaricus blazei Murrill by submerged fermentation. J. Microbiol., Immunol. Infect., 39(2):98-108.

[12]Liu, G.Q., Zhang, K.C., 2005. Mechanisms of the anticancer action of Ganoderma lucidum (Leyss. ex Fr.) Karst.: a new understanding. J. Integr. Plant Biol., 47(2):129-135.

[13]Liu, G.Q., Lin, Q.L., Ren, G.P., et al., 2010. Use of response surface methodology to optimize critical medium components for biomass and extracellular polysaccharide production by Ganoderma sinense. Afr. J. Microbiol. Res., 4(24):2735-2741.

[14]Liu, G.Q., Wang, X.L., Han, W.J., et al., 2012. Improving the fermentation production of the individual key triterpene ganoderic acid me by the medicinal fungus Ganoderma lucidum in submerged culture. Molecules, 17(11):12575-12586.

[15]Liu, R.M., Zhong, J.J., 2011. Ganoderic acid Mf and S induce mitochondria mediated apoptosis in human cervical carcinoma HeLa cells. Phytomedicine, 18(5):349-355.

[16]Ma, K., Ren, J., Han, J., et al., 2014. Ganoboninketals A-C, antiplasmodial 3,4-seco-27-norlanostane triterpenes from Ganoderma boninense Pat. J. Nat. Prod., 77(8):1847-1852.

[17]Mao, X.B., Eksriwong, T., Chauvatcharin, S., et al., 2005. Optimization of carbon source and carbon/nitrogen ratio for cordycepin production by submerged cultivation of medicinal mushroom Cordyceps militaris. Process Biochem., 40(5):1667-1672.

[18]Min, B.S., Nakamura, N., Miyashiro, H., et al., 1998. Triterpenes from the spores of Ganoderma lucidum and their inhibitory activity against HIV-1 protease. Chem. Pharm. Bull., 46(10):1607-1612.

[19]Munck, L., Jorgensen, K.G., Ruud-Hansen, J., et al., 1989. The EBC methods for determination of high molecular weight β-glucan in barley, malt, wort and beer. J. I. Brewing, 95(2):79-82.

[20]Nishitoba, T., Sato, H., Kasai, T., et al., 1984. New bitter C27 and C30 terpenoids from the fungus Ganoderma lucidum (Reishi). Agric. Biol. Chem., 48(11):2905-2907.

[21]Paterson, R.R.M., 2006. Ganoderma—a therapeutic fungal biofactory. Phytochemistry, 67(18):1985-2001.

[22]Qiao, Y., Zhang, X.M., Qiu, M.H., 2007. Two novel lanostane triterpenoids from Ganoderma sinense. Molecules, 12(8):2038-2046.

[23]Shiao, M.S., 2003. Natural products of the medicinal fungus Ganoderma lucidum: occurrence, biological activities, and pharmacological functions. Chem. Rec., 3(3):172-180.

[24]Tang, W., Liu, J.W., Zhao, W.M., et al., 2006. Ganoderic acid T from Ganoderma lucidum mycelia induces mitochondria mediated apoptosis in lung cancer cells. Life Sci., 80(3):205-211.

[25]Tang, Y.J., Zhong, J.J., 2002. Fed-batch fermentation of Ganoderma lucidum for hyperproduction of polysaccharide and ganoderic acid. Enzyme Microb. Technol., 31(1):20-28.

[26]Tang, Y.J., Zhong, J.J., 2004. Modeling the kinetics of cell growth and ganoderic acid production in liquid static cultures of the medicinal mushroom Ganoderma lucidum. Biochem. Eng. J., 21(3):259-264.

[27]Tang, Y.J., Zhang, W., Zhong, J.J., 2009. Performance analyses of a pH-shift and DOT-shift integrated fed-batch fermentation process for the production of ganoderic acid and Ganoderma polysaccharides by medicinal mushroom Ganoderma lucidum. Bioresource Technol., 100(5):1852-1859.

[28]Thyagarajan, A., Jedinak, A., Nguyen, H., et al., 2010. Triterpenes from Ganoderma Lucidum induce autophagy in colon cancer through the inhibition of p38 mitogen-activated kinase (p38 MAPK). Nutr. Cancer, 62(5):630-640.

[29]Wagner, R., Mitchell, D.A., Lanzi Sassaki, G., et al., 2003. Current techniques for the cultivation of Ganoderma lucidum for the production of biomass, ganoderic acid and polysaccharides. Food Technol. Biotechnol., 41(4):371-382.

[30]Wagner, R., Mitchell, D.A., Sassaki, G.L., et al., 2004. Links between morphology and physiology of Ganoderma lucidum in submerged culture for the production of exopolysaccharide. J. Biotechnol., 114(1):153-164.

[31]Wasser, S.P., 2010. Medicinal mushroom science: history, current status, future trends, and unsolved problems. Int. J. Med. Mushrooms, 12(1):1-16.

[32]Wu, G.S., Lu, J.J., Guo, J.J., et al., 2012. Ganoderic acid DM, a natural triterpenoid, induces DNA damage, G1 cell cycle arrest and apoptosis in human breast cancer cells. Fitoterapia, 83(2):408-414.

[33]Xiao, J.H., Chen, D.X., Wan, W.H., et al., 2006. Enhanced simultaneous production of mycelia and intracellular polysaccharide in submerged cultivation of Cordyceps jiangxiensis using desirability functions. Process Biochem., 41(8):1887-1893.

[34]Xu, J.W., Zhao, W., Zhong, J.J., 2010. Biotechnological production and application of ganoderic acids. Appl. Microbiol. Biotechnol., 87(2):457-466.

[35]Xu, P., Ding, Z.Y., Qian, Z., et al., 2008. Improved production of mycelial biomass and ganoderic acid by submerged culture of Ganoderma lucidum SB97 using complex media. Enzy. Microb. Technol., 42(4):325-331.

[36]Yang, M., Wang, X., Guan, S., et al., 2007. Analysis of triterpenoids in Ganoderma lucidum using liquid chromatography coupled with electrospray ionization mass spectrometry. J. Am. Soc. Mass Spectrom., 18(5):927-939.

[37]Zhang, J., Zhang, L., Duan, J., et al., 2006. On-line hyphenation of supercritical fluid extraction and two-dimensional high performance liquid chromatography-atmospheric pressure chemical ionization tandem mass spectrometer for the analysis of Ganoderma lucidum. J. Sep. Sci., 29(16):2514-2522.

[38]Zhang, W.X., Zhong, J.J., 2010. Effect of oxygen concentration in gas phase on sporulation and individual ganoderic acids accumulation in liquid static culture of Ganoderma lucidum. J. Biosci. Bioeng., 109(1):37-40.

[39]Zhao, W., Xu, J.W., Zhong, J.J., 2011. Enhanced production of ganoderic acids in static liquid culture of Ganoderma lucidum under nitrogen-limiting conditions. Bioresour. Technol., 102(17):8185-8190.

Open peer comments: Debate/Discuss/Question/Opinion

<1>

Please provide your name, email address and a comment





Journal of Zhejiang University-SCIENCE, 38 Zheda Road, Hangzhou 310027, China
Tel: +86-571-87952783; E-mail: cjzhang@zju.edu.cn
Copyright © 2000 - 2024 Journal of Zhejiang University-SCIENCE