樟芝(Antrodia cinnamomea)是一种珍稀食药两用蕈菌,具有解酒保肝、抗病毒、抗肿瘤、调节免疫等多种生物活性。基于无性孢子接种的深层发酵技术是樟芝目前最高效、最普遍的人工培养方式,但樟芝深层发酵产孢量低,种子制备成本高,限制了樟芝深层发酵工业化生产效率及效益。为了提高樟芝深层发酵产孢效率,该文首先考察了牛樟树不同溶剂(甲醇、乙酸乙酯、石油醚及水)提取物对樟芝深层发酵无性产孢的影响,发现只有牛樟树水提物能显著促进樟芝无性产孢,且在最佳添加量(60 μg/mL)下,可使樟芝产孢量较对照组提高58%左右,同时生物量提高26%左右,三萜产量提高35%左右,多糖产量提高270%左右(即产量是原来的3.7倍);随后,该文将添加牛樟树水提物促进产生的樟芝孢子接种发酵,发现其产孢性能及产胞内多糖性能均显著优于对照组(不添加牛樟树水提物发酵产生的樟芝孢子),其中最大产孢量及胞内多糖产量分别提高了28%及46%左右。该研究成果操作方便、成本低廉、效果显著,较具开发价值及应用前景。
Antrodia cinnamomea (syn Antrodia camphorata and Taiwanofungus camphoratus) is a kind of valuable edible and medicinal mushroom, and has various biological activities such as hangover alleviation, hepatoprotective, anti-virus, anti-tumor, and regulating immunity. Nevertheless, wild A. cinnamomea fruiting bodies are scarce and in short supply. Thus, the submerged fermentation based on spore inoculation is currently the most efficient artificial cultivation method for A. cinnamomea. However, the submerged fermentation of A. cinnamomea still has some disadvantages, such as low spore production and high cost of inoculum preparation, which largely limit its industrial production efficiency and benefits. The effects of methanol extract, ethyl acetate extract, petroleum ether extract, and water extract of Cinnamomum kanehirae Hay on the sporulation of A. cinnamomea in submerged fermentation were compared, and only the water extract of C. kanehirae (WEC) significantly promoted the spore production of A. cinnamomea. When 60 μg/mL of WEC was added to the medium, the spore production increased by 58% compared with the control. At the same time, the biomass and total triterpene increased by 26% and 35%, respectively. The crude intercellular polysaccharide production increased to 3.7 times of the control (i.e. increased by 270%). Lastly, the spores of A. cinnamomea induced by WEC as inoculum were used to study its fermentation performance. The sporulation performance and intracellular polysaccharide production of spores induced by WEC were significantly better than that without WEC inducing (control). Specifically, the maximum spore production and the intracellular polysaccharide production of spores increased by 28% and 46%, respectively. This method has the advantages of convenience, low-cost, and high efficacy, which has great development value and good application prospects.
[1] WU S H, KIRK P M, REDHEAD S A, et al.Resolution of the nomenclature for niu-chang-chih (Taiwanofungus camphoratus), an important medicinal polypore[J].Taxon, 2012, 61(6):1 305-1 310.
[2] 陈稳竹, 钟建荣, 杨善岩.牛樟芝的研究现状·问题与展望[J].安徽农业科学, 2018, 46(27):25-28;53.
CHEN W Z, ZHONG J R, YANG S Y.Research advances, issues and persprctives of Taiwanofungus camphorates[J].Journal of Anhui Agricultural Sciences, 2018, 46(27):25-28;53.
[3] 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].Oncotargets and Therapy, 2014, 8:99-109.
[4] HSIAO H Y.Hepatoprotective effect of an Antrodia cinnamomea product via a novel process on carbon tetrachloride-induced hepatotoxicity in rats[J].International Journal of Nutrition and Food Sciences, 2018, 7(4):134.
[5] CHEN Y F, LU W L, WU M D, et al.Analysis of Taiwan patents for the medicinal mushroom “Niu-Chang-Chih”[J].Recent Patents on Food, Nutrition & Agriculture, 2013, 5(1):62-69.
[6] REN Y L, LI H X, ZHOU L X, et al.Protective effect of spore powder of Antrodia camphorate ATCC 200183 on CCl 4-induced liver fibrosis in mice[J].Nutrients, 2020, 12(9):2 778.
[7] 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):1 332-1 345.
LI H X, LU Z M, GENG Y, et al.Recent advance in submerged fermentation of Taiwanofungus camphoratus[J].Mycosystema, 2017, 36(10):1 332-1 345.
[8] 韩金龙, 张蕾, 毛建卫, 等.樟芝深层发酵及活性物质三萜提取工艺的研究[C].中国食品科学技术学会第十一届年会论文摘要集,2014:333-335.
HAN J L, ZHANG L, MAO J W, et al.Research of submerged fermentation and extract of activated substance triterpenoids from Antrodia camphorate[C].The 11th Annual Meeting of CIFST, 2014:333-335.
[9] 张知晓, 季梅, 泽桑梓.牛樟芝培养技术的研究进展[J].热带农业科学, 2015, 35(3):94-99.
ZHANG Z X, JI M, ZE S Z.Research advances in Antrodia camphorata cultivation technology[J].Chinese Journal of Tropical Agriculture, 2015, 35(3):94-99.
[10] 姚秀英. 牛樟芝人工培养条件的初步研究[D].济南:山东师范大学, 2011.
YAO X Y.The preliminary study on artificial cultivation condition of Taiwanofungus camphoratus[D].Jinan:Shandong Normal University, 2011.
[11] GENG Y, HE Z, LU Z M, et al.Antrodia camphorata ATCC 200183 sporulates asexually in submerged culture[J].Applied Microbiology and Biotechnology, 2013, 97(7):2 851-2 858.
[12] LU Z M, LEI J Y, XU H Y, et al.Optimization of fermentation medium for triterpenoid production from Antrodia camphorata ATCC 200183 using artificial intelligence-based techniques[J].Applied Microbiology and Biotechnology, 2011, 92(2):371-379.
[13] 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.
[14] 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.
[15] SHEN Y C, CHOU C J, WANG Y H, et al.Anti-inflammatory activity of the extracts from mycelia of Antrodia camphorata cultured with water-soluble fractions from five different Cinnamomum species[J].FEMS Microbiology Letters, 2004, 231(1):137-143.
[16] CHANG T T, WANG W R.The role of four essential oils on mycelial growth and basidiomatal formation of Antrodia cinnamomea[J].Taiwan Journal of Forest Science, 2008, 23(1):105-110.
[17] LU Z M, GENG Y, LI H X, et al.Alpha-terpineol promotes triterpenoid production of Antrodia cinnamomea in submerged culture[J].FEMS Microbiology Letters, 2014, 358(1):36-43.
[18] 李一凡, 谢观雯, 李翁坤, 等.植物激素及木质素对牛樟芝菌丝体生长情况的影响[J].中草药, 2019, 50(6):1 453-1 460.
LI Y F, XIE G W, LI W K, et al.Effects of phytohormones and lignin on growth of Antrodia cinnamomea mycelium[J].Chinese Traditional and Herbal Drugs, 2019, 50(6):1 453-1 460.
[19] 李华祥. 樟芝无性孢子介导的高效循环发酵策略及其产孢分子机制解析[D].无锡:江南大学, 2017.
LI H X.Arthroconidia-mediated repeated batch fermentation strategy for Antrodia camphorate and underlying mechanism of asexual sporulation[D].Wuxi:Jiangnan University, 2017.
[20] 胡生辉, 刘君良, 徐国祺.樟树叶提取物对木材腐败菌的抑菌性研究[J].林业科技通讯, 2017(5):76-79.
HU S H, LIU J L, XU G Q.Inhibitory activity of Cinnamomum camphora leaves extracts on selected wood fungi and moulds[J].Forest Science and Technology, 2017(5):76-79.