Illigera aromatica, a medicinal liana, was fermented with Clonostachys rogersoniana. Two novel menthane-type monoterpenoid dimers, dimericilligerates E (1) and F (2) were isolated from the fermented material. Their structures were identified by 1D/2D NMR, electric circular dichroism, derivatization, and Mosher's method. A novel elimination reaction that formed a benzene ring from a menthane moiety afforded derivative 1a, which possesses more resolved NMR signals than the dimeric monoterpenoid, dimericilligerate E (1). The determination of 1a verified the estimation of 1. Compounds 1 and 2 showed potent cytotoxic activities; especially, 1 possessed significant activities against the liver hepatocellular carcinoma cell line (SMMC-7721) with an IC50 value of 3.89 ± 0.23 μM. Three novel precursors, dimericilligerates A–C (3–5), were identified by HPLC-MS and isolated from the original I. aromatica. This paper presents a novel class of monoterpenoid dimers and suggests that C. rogersoniana-fermented I. aromatica is effective to produce cytotoxic dimeric monoterpenoids from inactive original materials.

•This is the first to report that pure strain fermentation processing of I. henryi.•C. rogersoniana is an effective strain for fermentation of I. henryi.•The AChE inhibitory activity of fermented I. henryi was improved significantly.•A key biotransformation of (S)-actinodaphnine (1) to a new alkaloid 2 was found.•New produced alkaloid 2 possessed stronger AChE inhibitory effect.Illigera henryi, an endemic traditional Chinese medicine, contains abundant aporphine alkaloids that possess various bioactivities. In the present study, tubers of I. henryi were fermented by several fungi, and the acetylcholinesterase (AChE) inhibitory activities of non-fermented and fermented I. henryi were measured. The results showed that the fermentation of I. henryi with Clonostachys rogersoniana 828H2 is effective for improving the AChE inhibitory activity. A key biotransformation was found during the C. rogersoniana fermentation for clarifying the improvement of the AChE inhibitory activity of I. henryi: (S)-actinodaphnine (1) was converted to a new 4-hydroxyaporphine alkaloid (4R,6aS)-4-hydroxyactinodaphnine (2) that possessed a stronger AChE inhibitory activity, with an IC50 value of 17.66 ± 0.06 μM. This paper is the first to report that the pure strain fermentation processing of I. henryi and indicated C. rogersoniana fermentation might be a potential processing method for I. henryi.

The antioxidant activities and total phenolic content of fermented Salvia miltiorrhiza with fungus Geomyces luteus were investigated. The results revealed that G. luteus fermentation could significantly improve the antioxidant activity and total phenolic content of S. miltiorrhiza. The main antioxidant constituents were characterized by spectroscopic analysis as salvianolic acids. High-performance liquid chromatography (HPLC) quantification also showed the enhanced content of salvianolic acid B after fermentation. The present study suggests that G. luteus fermentations are effective in the S. miltiorrhiza salvianolic acids’ enrichment process.研究真菌固态发酵对丹参抗氧化活性的影响及探 索丹酚酸富集的途径。该研究发现真菌G. luteus 固态发酵可增加丹参的 抗氧化活性及有效地富集丹酚酸B,为丹参更有 效的开发利用提供了新思路。采用20 种真菌对丹参进行固态发酵,以两种抗 氧化模型(二苯代苦味酰基自由基(DPPH•)和 2,2'-联氮基双(3-乙基苯并噻唑啉-6-磺酸)二铵 盐自由基(ABTS+•))测定发酵物的抗氧化活性; 采用Folin-Ciocaltea 比色法测定发酵物的总酚含 量;以多种波谱学方法(紫外光谱(UV)、红外 光谱(IR)和核磁共振波谱(NMR))和高效液 相色谱法(HPLC)表征发酵前后丹参化学成分 的变化。抗氧化活性测试和总酚测定结果表明,G. luteus 发酵可有效地增加丹参的抗氧化活性和总酚含 量。UV、IR 及NMR 等波谱表征发现,在丹参经 G. luteus 发酵后,丹酚酸类化合物的含量提高了; 同时HPLC 测定也发现,丹酚酸B含量显著增加。 这些结果表明,G. luteus 发酵可实现丹酚酸B 的 富集。