基于NMR特征的云南咖啡活性化学成分研究

	基于NMR特征的云南咖啡活性化学成分研究
	洪德福
导师	邱明华
关键词	二萜，导向分离，α-葡萄糖苷酶，分子对接，发酵 Diterpenes, Targeted separation, α-Glucosidase, Molecular docking, Fermentation
摘要	咖啡作为世界著名的饮料之一，深受各国消费者的喜爱。云南作为全国最大的咖啡种植和出口基地，云南主要以栽种小粒咖啡 (Coffea arabica) 为主，可分为波旁属、铁皮卡属和卡帝姆 (Catimor) 等。此前研究多集中于杂交品种卡帝姆，其他品种未进行过系统的植物化学研究。因此，本研究以云南栽种的另一个杂交咖啡品种S288为研究对象，利用本实验建立的咖啡二萜数据库和开发的DATAanalyte软件对二萜化学成分进行导向分离，结合多种色谱分离手段进行单体纯化和波谱鉴定，从中共分离鉴定了60个化合物，包括2种类型二萜新骨架和32个新化合物。Cafespirone acid A (1) 和Cafespirone acid B (2) 是一种由6/6/5三环构成的螺环骨架的二萜；Cafeane acid B (3)、Cafeane acid A (4) 是由于C/D环重排，从而形成一个6/6/6/5四环结构二萜；这2种类型二萜尚未见报道；此外，分离鉴定的化合物分别为: 20位迁移 (10→9) 对映-贝壳杉烷型二萜 (5-10)，阿替生烷型二萜 (11和12) 对映-滨海孪生花烷型二萜 (14-18)，氧化型对映-贝壳杉烷型二萜 (19-26、48和49)，γ-内酯型对映-贝壳杉烷型二萜 (27-38)，内酰胺型对映-贝壳杉烷型二萜 (39-42)，呋喃型对映-贝壳杉烷型二萜 (43-47)；降解型对映-贝壳杉烷型二萜 (50-56)，松香烷型二萜 (57) 为首次在咖啡中发现，苯并A环型的对映-贝壳杉烷型二萜 (58)，裂解型的对映-贝壳杉烷型二萜 (59和60)。对分离鉴定的部分二萜化合物 (1、2、4、5、7、8、14-17、21、23、24、31、32、34、48、50、57) 进行α-葡萄糖苷酶抑制活性测定。结果表明，化合物4、5、8、17、21、31、32、34和48显示优于阿卡波糖中等的抑制活性，并通过分子对接进行构型关系讨论。云南得天独厚的自然条件，海拔和纬度造就了咖啡豆酸味适中、浓而不苦、香味醇和、略带果味的特质，但与瑰夏 (Geisha) 等精品咖啡相比，存在花果甜香风味有所欠缺。咖啡风味不仅受品种和种植环境的影响，采收后加工过程中发酵环节也起到至关重要的作用。因此，本研究选用蛋白酶和果胶酶对云南咖啡进行单酶厌氧发酵。结果表明，通过发酵之后，咖啡品质得到明显提升，均达到精品咖啡评分，同时增加了咖啡其他风味。进一步改善和提高云南咖啡风味提供一定理论依据和数据支持。; Coffee is one of the most famous drinks in the world. It is deeply loved by consumers all over the world. Yunnan is the largest coffee planting and exporting base in the country. It mainly cultivates Coffea arabica, which can be divided into Bourbon, Typica and Catimor. Previous studies have focused on the hybrid Catimor, and no systematic phytochemical studies have been performed on other species. Therefore, in this study, another hybrid coffee variety S288 grown in Yunnan as the research object.; the coffee diterpene database established by our research group and the developed DATAanalyte software were used to conduct the directional separation of diterpene chemical components, and a variety of chromatographic separation methods were used to carry out single-phase separation. Through body purification and spectral identification, 60 compounds were isolated and identified, including 2 new skeletons of diterpenes and 32 new compounds. Cafespirone acid A (1) and Cafespirone acid B (2) are diterpenes with a spiro-ring skeleton composed of 6/6/5 tricyclic rings; Cafeane acid B (3), Cafeane acid A (4) are possesses a 6/6/6/5 tetracyclic system due to the C/D ring rearrangement; These two types of diterpenes have not been reported; in addition, the isolated and identified compounds are: rearranged-type diterpenes (5-10), atisane-type diterpenes (11 and 12), Villanova-type diterpenes (14-18), oxidized-type diterpenes (19-26, 48, and 49), γ-lactone-type diterpenes (27-38), lactam-type diterpenes (39-42)，furan-type diterpenes (43-47); abietane-type diterpene (57), first discovered in coffee, benzo-A ring-type diterpenes (58), lysis-type diterpenes (59 and 60). Part of the diterpenoids (1, 2, 4, 5, 7, 8, 14-17, 21, 23, 24, 31, 32, 34, 48, 50, 57) isolated and identified were evaluated for their α-glucosidase inhibitory activity. The results showed that compounds 4, 5, 8, 17, 21, 31, 32, 34 and 48 had a moderate inhibitory effect on α-glucosidase, and the configurational relationship is discussed by molecular docking. Yunnan's unique natural conditions, altitude and latitude make coffee beans moderately sour, strong but not bitter, mellow and slightly fruity. However, compared with specialty coffees such as Geisha, there is a lack of sweet and fruity flavors. The flavor of coffee is not only affected by the variety and planting environment, but also the fermentation process during post-harvest processing also plays a crucial role. Therefore, in this study, protease and pectinase were used for single-enzyme anaerobic fermentation of Yunnan Coffee. The results showed that after fermentation, the coffee quality was significantly improved, all of which reached the specialty coffee score, and at the same time imparted other flavors to the coffee. Further improvement and enhancement of Yunnan coffee flavor provides certain theoretical basis and data support.
语种	中文
	2022-05
学位授予单位	中国科学院大学
文献类型	学位论文
条目标识符	http://ir.kib.ac.cn/handle/151853/75137
专题	昆明植物所硕博研究生毕业学位论文
推荐引用方式 GB/T 7714	洪德福. 基于NMR特征的云南咖啡活性化学成分研究[D]. 中国科学院大学,2022.

条目包含的文件
文件名称/大小	文献类型	版本类型	开放类型	使用许可
洪德福-洪德福-201928010642（14211KB）	学位论文		限制开放	CC BY-NC-SA	请求全文