Identification and application of effectors promoting asexual sporulation of Antrodia cinnamomea in aqueous extract of Cinnamomum kanehirae Hay

Abstract

Abstract: In order to determine the substance in aqueous extract of Cinnamomum kanehirae Hay (CWE) that promotes the asexual sporulation of Antrodia cinnamomea in submerged fermentation. Firstly, the CWE was isolated by alcohol precipitation and fractional extraction with different organic solvents, and the sporulation was used as the index to study the influence of different components on the sporulation of A. cinnamomea. It was showed that 30 μg/mL of chloroform extract of supernatant from alcohol precipitation of CWE (LFE) and 50 μg/mL of ethyl acetate extract (YZE) had a remarkable effect on promoting sporulation of A. cinnamomea, indicating that the effectors mainly existed in LFE and YZE; Subsequently, LC-MS/MS was used to analyze the components of different extracts of CWE, and combined with their effects on sporulation of A. cinnamomea, it was judged that the effector was probably erythritol; Finally, the commercial erythritol with purity of 98% was used to verify its effect on sporulation of A. cinnamomead. It was showed that erythritol did significantly promote the sporulation of A. cinnamomea, and the sporulation increased by 55.17% compared with Control at the optimal concentration of 1 μg/mL. Meanwhile, 1 μg/mL of erythritol can also significantly promote the growth and the synthesis of intracellular polysaccharides (IPS) for A. cinnamomea in submerged fermentation, and the biomass and yield of IPS were increased by 18.65% and 260.13%, respectively. However, erythritol has no significant effect on the synthesis of triterpenes. This study can greatly improve the industrial production efficiency and benefits of submerged fermentation for A. cinnamomea, and have great development value and good application prospects.

Key words: Antrodia cinnamomea, submerged fermentation, asexual sporulation, Cinnamomum kanehirae Hay, erythritol, liquid chromatography-mass spectrometry (LC-MS)

CLC Number:

TS201.3

Hua-Xiang LI. Identification and application of effectors promoting asexual sporulation of Antrodia cinnamomea in aqueous extract of Cinnamomum kanehirae Hay[J]. FOOD SCIENCE.

References

[1] CHEN C L, LI, W C, CHUANG, Y C, et al. Sexual crossing, chromosome-level genome sequences, and comparative genomic analyses for the medicinal mushroom Taiwanofungus camphoratus (Syn. Antrodia Cinnamomea, Antrodia Camphorata) [J]. Microbiology Spectrum, 2022, 10: e02032-21. DOI: 10.1128/spectrum.02032-21.
[2] LU M C, EL-SHAZLY M, WU T Y, et al. Recent research and development of Antrodia cinnamomea[J]. Pharmacology & Therapeutics, 2013, 139: 124-156. DOI: 10.1016/j.pharmthera.2013.04.001.
[3] XU X Y, GENG Y, XU H X, et al. Antrodia camphorata-derived antrodin C inhibits liver fibrosis by blocking TGF-Beta and PDGF signaling pathways[J]. Frontiers in Molecular Biosciences, 2022, 9: 835508. DOI:10.3389/fmolb.2022.835508.
[4] HUANG T F, WANG S W, LAI Y W, et al. 4-Acetylantroquinonol B suppresses prostate cancer growth and angiogenesis via a VEGF/PI3K/ERK/mTOR-dependent signaling pathway in subcutaneous xenograft and in vivo angiogenesis Models[J]. International Journal of Molecular Sciences, 2022, 23: 1446. DOI: 10.3390/ijms23031446.
[5] 李华祥, 石瑀, 蒋文浩, 等. 樟芝深层发酵菌丝体提取物对食源性致病菌的抑制作用[J]. 中国食品学报, 2022, 22(3): 47-52. DOI: 10.16429/j.1009-7848.2022.03.006.
[6] LU C L, LI H X, ZHU X Y, et al. Regulatory effect of intracellular polysaccharides from Antrodia cinnamomea on the intestinal microbiota of mice with antibiotic-associated diarrhea[J]. Quality Assurance and Safety of Crops & Foods, 2022, 14: 124-134. DOI: 10.15586/qas.v14i3.1073.
[7] LU C L, LEE B H, REN Y L, et al. Effects of exopolysaccharides from Antrodia cinnamomea on inflammation and intestinal microbiota disturbance induced by antibiotics in mice[J]. Food Bioscience, 2022, 50: 102116. DOI: 10.1016/j.fbio.2022.102116.
[8] LI H X, WANG J J, LU C L, et al. Review of bioactivity, isolation, and identification of active compounds from Antrodia cinnamomea[J]. Bioengineering, 2022, 9: 494. DOI: 10.3390/bioengineering9100494.
[9] 李华祥, 陆震鸣, 耿燕, 等. 樟芝深层发酵工艺研究进展[J]. 菌物学报, 2017, 36(10): 1332-1345. DOI: 10.13346/j.mycosystema.170018.
[10] LU Z M, HE Z, LI H X, et al. Modified arthroconidial inoculation method for the efficient fermentation of Antrodia camphorata ATCC 200183[J]. Biochemical Engineering Journal, 2014, 87: 41-49. DOI: 10.1016/j.bej.2014.03.020.
[11] LI H X, LU Z M, GENG Y, et al. Efficient production of bioactive metabolites from Antrodia camphorata ATCC 200183 by asexual reproduction-based repeated batch fermentation[J]. Bioresource Technology, 2015, 194: 334-343. DOI: 10.1016/j.biortech.2015.06.144.
[12] LI H X, DAI J N, SHI Y, et al. Molecular regulatory mechanism of the iron-ion-promoted asexual sporulation of Antrodia cinnamomea in submerged fermentation revealed by comparative transcriptomics[J]. Journal of Fungi, 2023, 9: 235. DOI: 10.3390/jof9020235.
[13] 李华祥, 陆震鸣, 朱青, 等. 钙离子调控樟芝深层发酵无性产孢及其分子机制[J]. 生物工程学报, 2017, 33(07): 1124-1135. DOI: 10.13345/j.cjb.170001.
[14] LI H X, LU Z M, ZHU Q, et al. Comparative transcriptomic and proteomic analyses reveal a flug-mediated signaling pathway relating to asexual sporulation of Antrodia camphorata[J]. Proteomics, 2017, 17: 1700256. DOI: 10.1002/pmic.201700256.
[15] LI H X, JI D, LUO Z S, et al. Comparative transcriptomic analyses reveal the regulatory mechanism of nutrient limitation-induced sporulation of Antrodia cinnamomea in submerged fermentation[J]. Foods, 2022, 11: 2715. DOI: 10.3390/foods11172715.
[16] 李华祥, 王娟娟, 石瑀, 等．金属离子促进樟芝深层发酵无性产孢[J]．食品与发酵工业, 2023, 49 (10) : 55-63. DOI: 10.13995/j. cnki.11-1802 /ts.033127.
[17] WU S H, KIRK P M, REDHEAD S A, et al. Resolution of the no-menclature for niu-chang-chih (Taiwanofungus camphoratus), an important medicinal polypore[J]. Taxon, 2012, 61(6): 1305-1310. DOI: 10.1002/tax.616011.
[18] 石瑀, 张雨佳, 任怡琳, 等. 牛樟树提取物对食源性致病菌的抑制效果[J]. 食品科技, 2021, 46(04): 168-172+179. DOI: 10.13684/j.cnki.spkj.2021.04.026.
[19] YEH R Y, SHIU Y L, SHEI S C, et al. Evaluation of the antibacterial activity of leaf and twig extracts of stout camphor tree, Cinnamomum kanehirae, and the effects on immunity and disease resistance of white shrimp, Litopenaeus vannamei[J]. Fish & Shellfish Immunology, 2009, 27(1): 26-32. DOI: 10.1016/j.fsi.2008.11.008.
[20] LIU Y K, CHEN K H, LEU Y L, et al. Ethanol extracts of Cinnamomum kanehirai Hayata leaves induce apoptosis in human hepatoma cell through caspase-3 cascade[J]. Onco Targets and Therapy, 2015, 8: 99-109. DOI: 10.2147/ott.s68765.
[21] ZHANG Z, WANG Y, YUAN X L, et al. Effects of culture mechanism of Cinnamomum kanehirae and C. camphora on the expression of genes related to terpene biosynthesis in Antrodia cinnamomea[J]. Mycobiology, 2022, 50(2): 121-131. DOI: 10.1080/12298093.2022.2059156.
[22] ZENG W W, CHEN T C, LIU C H. Identification and isolation of an intermediate metabolite with dual antioxidant and anti-proliferative activity present in the fungus Antrodia cinnamomea cultured on an alternative medium with Cinnamomum kanehirai leaf extract[J]. Plants (Basel), 2021, 10(4): 737. DOI: 10.3390/plants10040737.
[23] HSU F L, CHOU C J, CHANG Y C, et al. Promotion of hyphal growth and underlying chemical changes in Antrodia camphorata by host factors from Cinnamomum camphora[J]. International Journal of Food Microbiology, 2006, 106(1): 32-38. DOI: 10.1016/j.ijfoodmicro.2005.07.003.
[24] LU Z M, GENG Y, LI H X, et al. Alpha-terpineol promotes triterpenoid production of Antrodia cinnamomea in submerged culture[J]. FMS Microbiology Letters, 2014, 358(1): 36-43. DOI: 10.1111/1574-6968.12545.
[25] LI H T, YEH H C, CHEN C Y. Secondary metabolites of the leaves of Cinnamomum kanehirai[J]. Chemistry of Natural Compounds, 2016, 52(6): 1143-1144. DOI: 10.1007/s10600-016-1889-8.
[26] LIN C L, KAO C L, LI W J, et al. Secondary metabolites from the stems of Cinnamomum kanehirai[J]. Chemistry of Natural Compounds, 2018, 54(4): 762-763. DOI: 10.1007/s10600-018-2466-0.
[27] 李华祥, 石瑀, 罗志珊, 等. 牛樟树提取物促进樟芝深层发酵无性产孢及其应用[J]．食品与发酵工业, 2022, 48(16) : 65-71. DOI: 10.13995/j.cnki.11-1802 /ts.030352.
[28] ABDELHAFEZ O H, OTHMAN E M, FAHIM J R, et al. Metabolomics analysis and biological investigation of three Malvaceae plants[J]. Phytochemical Analysis, 2019, 31(2): 204-214. DOI: 10.1002/pca.2883.
[29] 李华祥, 吉丹, 陆春雷, 等. 樟芝深层发酵多糖对抗生素相关性腹泻小鼠肠道菌群的调节作用[J].食品科学, 2023, 44(13): 42-51. DOI: 1002-6630（2023）13-0042-10.
[30] YANG R X, GAO Z G, LIU X, et al. Root exudates from muskmelon (Cucumis melon. L) induce autotoxicity and promote growth of Fusarium oxysporum f. sp melonis[J]. Allelopathy Journal, 2013, 33(2): 175-188. DOI: <Go to ISI>://WOS:000335933300003.