振荡-静置循环培养新策略提升灵芝三萜合成与菌丝体活性
作者:
基金项目:

上海市现代农业产业技术体系项目(沪农科产字[2022]第 9 号);西藏自治区重点研发计划(XZ202301ZY0007N);上海市农业科学院卓越团队建设计划(2022A-03);上海市农业科学院攀高计划(2022–2024)


Oscillation-static cycle cultivation enhances the synthesis of triterpenes and mycelium activity in Ganoderma lucidum
Author:
  • JIANG Xingyi

    JIANG Xingyi

    Key Laboratory of Edible Fungi Resources and Utilization (South), Ministry of Agriculture and Rural Affairs, National Engineering Research Center of Edible Fungi, Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, Shanghai 201403, China;Shanghai Key Laboratory of New Drug Design, Engineering Research Centre of Pharmaceutical Process Chemistry, Ministry of Education, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China
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  • HAN Wei

    HAN Wei

    Shanghai Key Laboratory of New Drug Design, Engineering Research Centre of Pharmaceutical Process Chemistry, Ministry of Education, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China
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  • GUO Jia

    GUO Jia

    Key Laboratory of Edible Fungi Resources and Utilization (South), Ministry of Agriculture and Rural Affairs, National Engineering Research Center of Edible Fungi, Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, Shanghai 201403, China
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  • LIU Yanfang

    LIU Yanfang

    Key Laboratory of Edible Fungi Resources and Utilization (South), Ministry of Agriculture and Rural Affairs, National Engineering Research Center of Edible Fungi, Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, Shanghai 201403, China
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  • XU Aiguo

    XU Aiguo

    Xizang Plateau Institute of Biology, Lhasa 850000, Xizang, China
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  • TANG Chuanhong

    TANG Chuanhong

    Key Laboratory of Edible Fungi Resources and Utilization (South), Ministry of Agriculture and Rural Affairs, National Engineering Research Center of Edible Fungi, Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, Shanghai 201403, China
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  • FENG Jie

    FENG Jie

    Key Laboratory of Edible Fungi Resources and Utilization (South), Ministry of Agriculture and Rural Affairs, National Engineering Research Center of Edible Fungi, Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, Shanghai 201403, China
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  • ZHANG Jinsong

    ZHANG Jinsong

    Key Laboratory of Edible Fungi Resources and Utilization (South), Ministry of Agriculture and Rural Affairs, National Engineering Research Center of Edible Fungi, Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, Shanghai 201403, China
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  • 摘要
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  • 参考文献 [33]
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    摘要:

    灵芝作为历史悠久的食药两用型真菌,其药用价值主要源自灵芝三萜这一主要活性物质,灵芝三萜具有抗肿瘤、抗氧化等药理活性。本研究旨在建立高效液态发酵生产灵芝三萜的技术体系。针对传统深层发酵和振荡-静置两阶段培养的局限性,利用振荡-静置循环培养工艺,结合遗传算法构建人工神经网络模型进行优化。在优化条件下,液态发酵高产灵芝三萜的最佳培养方式为振荡2.8 d-静置7.3 d-振荡0.2 d-静置0.3 d。此条件下,灵芝三萜含量达到20.82 mg/g,灵芝酸得率为129.09 mg/L,与Z10J0相比提高了324.78%,且培养周期缩短至10.6 d。同时,该培养方式下菌丝体具有较好的抗肿瘤活性和抗氧化活性。本研究开发了一种经济有效的液态发酵培养方式,可简化工艺流程、缩短发酵周期并有效改善传统培养方式的弊端;同时为液态发酵高产灵芝三萜的规模化应用提供了参考,具有广阔的应用前景。

    Abstract:

    Ganoderma lucidum is a precious fungus with both edible and medicinal values and has a long history of medical use. Triterpenes as the main active components endow G. lucidum with anti-tumor, antioxidant, and other pharmacological activities. The present study endeavors to establish a proficient liquid-state fermentation technology for the enhanced production of triterpenes. In view of the limitations inherent in conventional submerged fermentation and oscillation-static two-stage cultivation, this study established an oscillation-static cycle cultivation process and optimized the cultivation conditions by building an artificial neural network model based on genetic algorithms. The cultivation conditions for the high-yield production of triterpenes were optimized as follows: 2.8 days of oscillation, 7.3 days of static cultivation, 0.2 day of oscillation, and 0.3 day of static cultivation. Under these conditions, the content of triterpenes reached 20.82 mg/g. The yield of triterpenes reached 129.09 mg/L, showing a remarkable increase of 324.78% compared with that of the Z10J0 method. Moreover, the established method shortened the cultivation cycle by 10.6 days. The mycelia cultivated under this regimen exhibited commendable anti-tumor and antioxidant activities. This study not only presents an economical liquid-state fermentation approach but also streamlines the fermentation flow, reduces fermentation duration, and effectively ameliorates drawbacks associated with conventional cultivation methods. In addition, this study gives valuable insights into the scaled application of liquid-state fermentation in the high-yield production of triterpenes, which showcases broad prospects.

    参考文献
    [1] 朱静, 师亮, 任昂, 刘锐, 赵明文. 灵芝三萜生物合成的研究进展[J]. 南京农业大学学报, 2022, 45(5): 981-989. ZHU J, SHI L, REN A, LIU R, ZHAO MW. Research progress on the biosynthesis of ganoderic acids[J]. Journal of Nanjing Agricultural University, 2022, 45(5): 981-989(in Chinese).
    [2] 唐庆九, 季哲, 郝瑞霞, 刘艳芳, 杨焱, 张劲松. 灵芝中性三萜类成分的抗肿瘤作用[J]. 食用菌学报, 2010, 17(1): 60-64. TANG QJ, JI Z, HAO RX, LIU YF, YANG Y, ZHANG JS. Inhibition of tumor cell proliferation by a neutral triterpenoid fraction from Ganoderma lucidum[J]. Acta Edulis Fungi, 2010, 17(1): 60-64(in Chinese).
    [3] MAU JL, LIN HC, CHEN CC. Antioxidant properties of several medicinal mushrooms[J]. Journal of Agricultural and Food Chemistry, 2002, 50(21): 6072-6077.
    [4] 王明宇, 刘强, 车庆明, 林志彬. 灵芝三萜类化合物对3种小鼠肝损伤模型的影响[J]. 药学学报, 2000, 35(5): 326-329. WANG MY, LIU Q, CHE QM, LIN ZB. Effects of triterpenoids from Ganoderma lucidum (leyss. ex Fr.) Karst on three different experimental liver injury models in mice[J]. Acta Pharmaceutica Sinica, 2000, 35(5): 326-329(in Chinese).
    [5] 彭珍华, 韩本勇, 赵声兰, 陈朝银. 三萜类化合物抗HIV研究进展[J]. 天然产物研究与开发, 2009, 21(B05): 258-262, 143. PENG ZH, HAN BY, ZHAO SL, CHEN CY. Research anti-HIV triterpenoid components[J]. Natural Product Research and Development, 2009, 21(B05): 258-262, 143(in Chinese).
    [6] XIA Q, ZHANG HZ, SUN XF, ZHAO HJ, WU LF, ZHU D, YANG GH, SHAO YY, ZHANG XX, MAO X, ZHANG LZ, SHE GM. A comprehensive review of the structure elucidation and biological activity of triterpenoids from Ganoderma spp.[J]. Molecules, 2014, 19(11): 17478-17535.
    [7] 姜沅彤. 灵芝主要活性成分的生物酶辅助提取及灵芝酸A药代动力学研究[D]. 长春: 吉林农业大学硕士学位论文, 2015. JIANG YT. Enzyme assisted extraction and ganoderic acid A pharmacokinetic study on main active components of Ganoderma lucidum[D]. Changchun: Master’s Thesis of Jilin Agricultural University, 2015(in Chinese).
    [8] ZHANG JM, ZHONG JJ, GENG AL. Improvement of ganoderic acid production by fermentation of Ganoderma lucidum with cellulase as an elicitor[J]. Process Biochemistry, 2014, 49(10): 1580-1586.
    [9] ZHONG JJ, XU YN, TAN GY, BAI LQ. Signal transduction engineering: a powerful platform technology for enhancing secondary metabolite production[J]. New Biotechnology, 2014, 31: S23-S24.
    [10] ZHOU XW, SU KQ, ZHANG YM. Applied modern biotechnology for cultivation of Ganoderma and development of their products[J]. Applied Microbiology and Biotechnology, 2012, 93(3): 941-963.
    [11] TANG YJ, ZHANG W, ZHONG JJ. 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[J]. Bioresource Technology, 2009, 100(5): 1852-1859.
    [12] LIU GQ, WANG XL, HAN WJ, LIN QL. Improving the fermentation production of the individual key triterpene ganoderic acid me by the medicinal fungus Ganoderma lucidum in submerged culture[J]. Molecules, 2012, 17(11): 12575-12586.
    [13] 毛健, 马海乐. 灵芝菌体液态深层发酵条件的优化[J]. 食品科学, 2009, 30(23): 377-382. MAO J, MA HL. Optimization of submerged fermentation of Ganoderma lucidum[J]. Food Science, 2009, 30(23): 377-382(in Chinese).
    [14] FANG QH, ZHONG JJ. Two-stage culture process for improved production of ganoderic acid by liquid fermentation of higher fungus Ganoderma lucidum[J]. Biotechnology Progress, 2002, 18(1): 51-54.
    [15] 邢源月, 王佳琦, 黄红梅. 灵芝深层液态发酵培养条件与营养条件优化[J]. 当代化工研究, 2022(10): 25-28. XING YY, WANG JQ, HUANG HM. Optimization of culture conditions and nutritional conditions for submerged liquid fermentation of Ganoderma lucidum[J]. Modern Chemical Research, 2022(10): 25-28(in Chinese).
    [16] 冯杰, 冯娜, 唐庆九, 颜梦秋, 杨焱, 周帅, 刘艳芳, 刘方, 张劲松. 补料方式对灵芝菌丝体液态深层发酵合成灵芝三萜的影响[J]. 食品科学, 2017, 38(12): 57-62. FENG J, FENG N, TANG QJ, YAN MQ, YANG Y, ZHOU S, LIU YF, LIU F, ZHANG JS. Effects of different feeding methods on production of triterpenes by Ganoderma lucidum in submerged fermentation[J]. Food Science, 2017, 38(12): 57-62(in Chinese).
    [17] 冯杰, 冯娜, 杨焱, 刘方, 贾薇, 张劲松. 通气量对灵芝菌丝体液态深层发酵合成灵芝三萜的影响[J]. 天然产物研究与开发, 2015, 27(9): 1564-1570. FENG J, FENG N, YANG Y, LIU F, JIA W, ZHANG JS. Effects of aeration rate on the production of triterpenes by Ganoderma lucidum G0119 in liquid submerged fermentation[J]. Natural Product Research and Development, 2015, 27(9): 1564-1570(in Chinese).
    [18] SINGHANIA RR, PATEL AK, SOCCOL CR, PANDEY A. Recent advances in solid-state fermentation[J]. Biochemical Engineering Journal, 2009, 44(1): 13-18.
    [19] OSMOLOVSKIY AA, BARANOVA NA, KREIER VG, KURAKOV AV, EGOROV NS. Solid-state and membrane-surface liquid cultures of micromycetes: specific features of their development and enzyme production (a review)[J]. Applied Biochemistry and Microbiology, 2014, 50(3): 219-227.
    [20] 王琼, 徐萌萌, 刘高强, 曹春蕾, 石贵阳, 丁重阳. 灵芝液体浅层静置培养高效生产三萜的研究[J]. 菌物学报, 2021, 40(3): 656-667. WANG Q, XU MM, LIU GQ, CAO CL, SHI GY, DING ZY. An efficient strategy to enhance triterpenoid production by liquid superficial-static culture (LSSC) of Ganoderma lingzhi[J]. Mycosystema, 2021, 40(3): 656-667(in Chinese).
    [21] 张忠, 张劲松, 刘艳芳, 周帅, 王金艳, 于华峥, 唐庆九. 分光光度法测定灵芝中总三萜含量方法探讨[J]. 上海农业学报, 2016, 32(1): 61-65. ZHANG Z, ZHANG JS, LIU YF, ZHOU S, WANG JY, YU HZ, TANG QJ. Discussion on the determination of total triterpenoids in Ganoderma lucidum by spectrophotometry[J]. Acta Agriculturae Shanghai, 2016, 32(1): 61-65(in Chinese).
    [22] 程池露, 汪旵, 王金艳, 唐传红, 张劲松, 冯娜, 唐庆九. 分光光度法测定灵芝三萜含量的干扰因素探讨[J]. 菌物学报, 2022, 41(10): 1716-1725. CHENG CL, WANG C, WANG JY, TANG CH, ZHANG JS, FENG N, TANG QJ. Discussion on interference factors in determination of triterpenes in Ganoderma lingzhi by spectrophotometric method[J]. Mycosystema, 2022, 41(10): 1716-1725(in Chinese).
    [23] XU JW, XU YN, ZHONG JJ. Production of individual ganoderic acids and expression of biosynthetic genes in liquid static and shaking cultures of Ganoderma lucidum[J]. Applied Microbiology and Biotechnology, 2010, 85(4): 941-948.
    [24] 邹立飞, 郑鹏. 人工神经网络和响应面法优化薏苡仁酒发酵条件[J]. 中国酿造, 2021, 40(1): 142-147. ZOU LF, ZHENG P. Optimization of fermentation conditions of coix seed wine by artificial neural network and response surface method[J]. China Brewing, 2021, 40(1): 142-147(in Chinese).
    [25] 郭嘉, 刘艳芳, 唐传红, 冯杰, 张劲松. 基于人工神经网络和遗传算法优化灵芝液态发酵培养基[J]. 食用菌学报, 2023, 30(3): 81-89. GUO J, LIU YF, TANG CH, FENG J, ZHANG JS. Optimization of Ganoderma lucidum liquid fermentation medium based on artificial neural network and genetic algorithm[J]. Acta Edulis Fungi, 2023, 30(3): 81-89(in Chinese).
    [26] 余素萍. 灵芝深层发酵生产生物活性物质的研究[D]. 南京: 南京农业大学硕士学位论文, 2004. YU SP. Study on submerged fermentation of ganoderma lucidum to produce bioactive substances[D]. Nanjing: Master’s Thesis of Nanjing Agricultural University, 2004(in Chinese).
    [27] LUO JG, LI L, KONG LY. Preparative separation of phenylpropenoid glycerides from the bulbs of Lilium lancifolium by high-speed counter-current chromatography and evaluation of their antioxidant activities[J]. Food Chemistry, 2012, 131(3): 1056-1062.
    [28] XIE J, SCHAICH KM. Re-evaluation of the 2,2-diphenyl-1-picrylhydrazyl free radical (DPPH) assay for antioxidant activity[J]. Journal of Agricultural and Food Chemistry, 2014, 62(19): 4251-4260.
    [29] 谭洪升, 李翔, 巩伯梁, 李刚. 灵芝子实体和孢子粉三萜含量的测定及体外抗肿瘤活性的评价[J]. 微生物学免疫学进展, 2018, 46(1): 43-48. TAN HS, LI X, GONG BL, LI G. Determination of total triterpenoids in fruiting body and spores of Ganoderma lucidum and assessment of their antitumor activity in vitro[J]. Progress in Microbiology and Immunology, 2018, 46(1): 43-48(in Chinese).
    [30] 李娜. 高产多糖和三萜的液态发酵专用灵芝优势菌株筛选及促三萜生成机制的探索[D]. 上海: 上海海洋大学硕士学位论文, 2021. LI N. Screening of dominant strains of Ganoderma lucidum for liquid fermentation with high production of polysaccharides and triterpenes and exploration of the mechanism of promoting triterpenoid production[D]. Shanghai: Master’s Thesis of Shanghai Ocean University, 2021(in Chinese).
    [31] 潘江安, 唐庆九, 冯娜, 唐传红, 王金艳, 刘艳芳, 袁峰, 徐国华, 颜梦秋, 谭贻, 张劲松, 周帅, 冯杰. 利用阶段pH控制提高灵芝深层发酵合成三萜能力的研究[J]. 食用菌学报, 2020, 27(2): 84-91. PAN JA, TANG QJ, FENG N, TANG CH, WANG JY, LIU YF, YUAN F, XU GH, YAN MQ, TAN Y, ZHANG JS, ZHOU S, FENG J. Enhanced triterpene biosynthesis in Ganoderma lucidum by staged pH control during submerged fermentation[J]. Acta Edulis Fungi, 2020, 27(2): 84-91(in Chinese).
    [32] TANG YJ, ZHONG JJ. Role of oxygen supply in submerged fermentation of Ganoderma lucidum for production of Ganoderma polysaccharide and ganoderic acid[J]. Enzyme and Microbial Technology, 2003, 32(3/4): 478-484.
    [33] 孙金旭. 溶氧控制条件对深层灵芝发酵生产灵芝酸产量的影响[J]. 北方园艺, 2014(5): 130-132. SUN JX. Effect of dissolved oxygen controlling conditions on production of ganoderic acids by Ganoderma[J]. Northern Horticulture, 2014(5): 130-132(in Chinese).
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引用本文

姜幸怡,韩伟,郭嘉,刘艳芳,徐爱国,唐传红,冯杰,张劲松. 振荡-静置循环培养新策略提升灵芝三萜合成与菌丝体活性[J]. 生物工程学报, 2024, 40(9): 3189-3200

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  • 收稿日期:2023-12-22
  • 最后修改日期:2024-03-25
  • 在线发布日期: 2024-09-24
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