真菌Stilbella sp. CGMCC 40422的萜类产物研究

doi:10.19586/j.2095-2341.2023.0030

生物技术进展 ›› 2023, Vol. 13 ›› Issue (4): 604-611.DOI: 10.19586/j.2095-2341.2023.0030

• 研究论文 • 上一篇

真菌Stilbella sp. CGMCC 40422的萜类产物研究

宋开南(), 艾羽桐, 徐玉泉()

中国农业科学院生物技术研究所，北京 100081

收稿日期:2023-03-14 接受日期:2023-05-06 出版日期:2023-07-25 发布日期:2023-08-03
通讯作者: 徐玉泉
作者简介:宋开南 E-mail: S15850653283@126.com；
基金资助:
国家自然科学基金项目(22277137)

Study of the Terpenoids from Stilbella sp. CGMCC 40422

Kainan SONG(), Yutong AI, Yuquan XU()

Biotechnology Research Institute，Chinese Academy of Agricultural Sciences，Beijing 100081，China

Received:2023-03-14 Accepted:2023-05-06 Online:2023-07-25 Published:2023-08-03
Contact: Yuquan XU

摘要/Abstract

摘要：

采用多种硅胶柱色谱、OSD柱色谱与半制备柱高效液相色谱对真菌Stilbella sp. CGMCC 40422大米培养基的发酵产物进行分离提纯，并通过质谱、核磁共振波谱解析等方法对得到的化合物纯品进行结构鉴定。结果显示，从该菌株中分离得到2个原萜烷型四环三萜、1个倍半萜与5个杂萜类化合物，分别为烟曲霉酸、helvolinic acid、tricho-acorenol、ascofuranone、ilicicolin C、LL-Z1272?、deacetylchloronectrin和ascochlorin N-acetylglucosamine。其中烟曲霉酸的含量最高，随后采用高效液相色谱法测定不同发酵时间发酵物中主产物烟曲霉酸的产量，烟曲霉酸含量在培养15 d后达最大值3 g·kg^-1，说明Stilbella sp. CGMCC 40422具有强大的萜类产物生产能力，研究结果为进一步开发萜类化合物生产的底盘细胞奠定了基础。

关键词: Stilbella sp.CGMCC40422, 萜类化合物, 烟曲霉酸, 底盘细胞

Abstract:

The compounds from Stilbella sp. CGMCC 40422 were isolated by silica gel column， ODS column and semi-preparative high performance liquid chromatography. And their structures were elucidated by nuclear magnetic resonance （NMR） and mass spectrum （MS） spectral analysis. The results showed that two prototerpene-type tetracyclic triterpenoids， one sesquiterpenoid and five meroterpenoids were isolated from the rice fermentation products of the fungus， and their structures were identified as helvolic acid， helvolinic acid， tricho-acorenol， ascofuranone， ilicicolin C， LL-Z1272?， deacetylchloronectrin ascochlorin and N-acetylglucosamine. The content of helvolic acid under different fermentation time was tested by HPLC chromatography. The yeild of helvolic acid reached a maximum of 3 g·kg^-1 after 15 days of fermentation， showing that Stilbella sp. CGMCC 40422 have strong production capacity of terpenoids. The results could lay the foundation for further development of the chassis cells of terpenoid cell factories.

Key words: Stilbella sp. CGMCC 40422, terpenoids, helvolic acid, chassis cells

中图分类号:

Q958.8

宋开南, 艾羽桐, 徐玉泉. 真菌Stilbella sp. CGMCC 40422的萜类产物研究[J]. 生物技术进展, 2023, 13(4): 604-611.

Kainan SONG, Yutong AI, Yuquan XU. Study of the Terpenoids from Stilbella sp. CGMCC 40422[J]. Current Biotechnology, 2023, 13(4): 604-611.

图/表 3

图1 菌株CGMCC 40422与近缘种的系统进化树

Fig. 1 Phylogenetic tree of strain CGMCC 40422 and related varieties

图2 化合物1~8的结构式注：1—烟曲霉酸；2—helvolinic acid；3—tricho-acorenol；4—ascofuranone；5—ilicicolin C；6—LL-Z1272?；7—deacetylchloronectrin；8—ascochlorin N-acetylglucosamine。图中C对应的数字见正文2.2对应化合物的相应参数。

Fig. 2 Structures of compounds 1~8

图3 烟曲霉酸产量的测定A：烟曲霉酸纯品的HPLC分析图谱（检测波长为230 nm）；B：粗提物浸膏的HPLC分析图谱；C：标准品的标准曲线；D：发酵时间对烟曲霉酸产量的影响

Fig. 3 Determination of helvolic acid yield

参考文献 26

1	BIAN G, DENG Z, LIU T. Strategies for terpenoid overproduction and new terpenoid discovery[J]. Curr. Opin. Biotechnol., 2017, 48: 234-241.
2	邓春萌,王哑倩,吴美媛.牛樟芝固态发酵菌丝体三萜及多糖提取工艺优化研究[J].生物技术进展,2020,10(3):328-331.
3	薛海洁,孙文涛,赵雨佳,等.微生物合成植物三萜及其皂苷化合物[J].生物产业技术,2019(1):19-26.
4	张春月,金佳杨,邱勇隽,等.传统与未来的碰撞:食品发酵工程技术与应用进展[J].生物技术进展,2021,11(4):418-429.
5	ZHAO Y, LV B, FENG X, et al.. Perspective on biotransformation and de vovo biosynthesis of licorice constituents[J]. J. Agric. Food Chem., 2017, 65(51): 11147-11156.
6	DAI Z, LIU Y, SUN Z, et al.. Identification of a novel cytochrome P450 enzyme that catalyzes the C-2α hydroxylation of pentacyclic triterpenoids and its application in yeast cell factories[J]. Metab. Eng., 2019, 51: 70-78.
7	GAO Q, WANG L, ZHANG M, et al.. Recent advances on feasible strategies for monoterpenoid production in Saccharomyces cerevisiae [J/OL]. Front. Bioeng. Biotechnol., 2020, 8: 609800[2020-12-01]. .
8	黄雪年,唐慎,吕雪峰.工业丝状真菌土曲霉合成生物技术研究进展及展望[J].合成生物学,2020,1(2):187-211.
9	VON D W, GODTFREDSEN W O, RASMUSSEN P R. Structure-activity relationships in fusidic acid-type antibiotics[J]. Adv. Appl. Microbiol., 1979, 25: 95-146.
10	LUO X, ZHOU X, LIN X, et al.. Antituberculosis compounds from a deep-sea-derived fungus Aspergillus sp. SCSIO Ind09F₀₁ [J]. Nat. Prod. Res., 2017, 31(16): 1958-1962.
11	蔡嘉慧,林夏,黄赛,等.海南霸王岭卷萼兜兰根际土壤真菌的分离与鉴定[J].现代农业科技,2020,23:113-115.
12	刘少华,陆金萍,朱瑞良,等.一种快速简便的植物病原真菌基因组DNA提取方法[J].植物病理学报,2005,35(4):362-365.
13	FUJIMOTO H, NEGISHI E, YAMAGUCHI K, et al.. Isolation of new tremorgenic metabolites from an ascomycete Corynascus setosus [J]. Chem. Pharm. Bull., 1996, 44(10): 1843-1848.
14	ZHU J, LIU L, WU M, et al.. Characterization of a sesquiterpene synthase catalyzing formation of cedrol and two diastereoisomers of tricho-acorenol from Euphorbia fischeriana [J]. J. Nat. Prod., 2021, 84(6): 1780-1786.
15	ZHANG P, BAO B, DANG H T, et al.. Anti-inflammatory sesquiterpenoids from a sponge-derived fungus Acremonium sp. [J]. J. Nat. Prod., 2009, 72(2): 270-275.
16	SINGH S B, BALL R G, BILLS G F, et al.. Chemistry and biology of cylindrols: novel inhibitors of ras farnesyl-protein transferase from Cylindrocarpon lucidum [J]. J. Org. Chem., 1996, 61(22): 7727-7737.
17	SEEPHONKAI P, ISAKA M, KITTAKOOP P, et al.. A novel ascochlorin glycoside from the insect pathogenic fungus Verticillium hemipterigenum BCC 2370[J]. J. Antibiot., 2004, 57(1): 10-16.
18	SUBKO K, KILDGAARD S, VICENTA F, et al.. Bioactive ascochlorin analogues from the marine-derived fungus Stilbella fimetaria [J/OL]. Mar. Drugs, 2021, 19(2): 46[2021-01-20]. .
19	LV J, HU D, GAO H, et al.. Biosynthesis of helvolic acid and identification of an unusual C-4-demethylation process distinct from sterol biosynthesis[J/OL]. Nat. Commun., 2017, 8(1): 1644[2017-11-21]. .
20	SANMANOCH W, MONGKOLTHANARUK W, KANOKMEDHAKUL S, et al.. Helvolic acid, a secondary metabolite produced by Neosartorya spinosa KKU-1NK1 and its biological activities[J]. Chiang Mai J. Sci., 2016, 43(43): 1-11.
21	XIAO J H, ZHANG Y, LIANG G Y, et al.. Synergistic antitumor efficacy of antibacterial helvolic acid from Cordyceps taii and cyclophosphamide in a tumor mouse model[J]. Exp. Biol. Med. (Maywood), 2017, 242(2): 214-222.
22	CHEN K, YUAN Y, WANG Z, et al.. Helvolic acid attenuates osteoclast formation and function via suppressing RANKL-induced NFATc₁ activation[J]. J. Cell. Physiol., 2019, 234(5): 6477-6488.
23	ARAKI Y, AWAKAWA T, MATSUZAKI M, et al.. Complete biosynthetic pathways of ascofuranone and ascochlorin in Acremonium egyptiacum [J]. Proc. Natl. Acad. Sci. USA, 2019, 116(17): 8269-8274.
24	GUTIÉRREZ M, THEODULOZ C, RODRÍGUEZ J, et al.. Bioactive metabolites from the fungus Nectria galligena, the main apple canker agent in Chile[J]. J. Agric. Food Chem., 2005, 53(20): 7701-7708.
25	徐帆,朱一翔,卢艳花.烟曲霉酸的发酵培养基优化和反应器放大研究[J].中国酿造,2017,36(4):131-136.
26	谭雁鸿,李基兴,林秀萍,等.中国南海软珊瑚真菌Eupenicillium sp.DX-SER3(KC871024)的次级代谢产物研究[J].热带海洋学报,2019,38(2):43-47.

真菌Stilbella sp. CGMCC 40422的萜类产物研究

Study of the Terpenoids from Stilbella sp. CGMCC 40422

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