Sarsaiya Surendra, Jain Archana, Shu Fuxing, Yang Mingfa, Pu Mengxuan, Jia Qi, Gong Qihai, Wu Qin, Qian Xu, Shi Jingshan, Chen Jishuang
Key Laboratory of Basic Pharmacology and Joint International Research Laboratory of Ethnomedicine of Ministry of Education, Zunyi Medical University, Zunyi, China.
Bioresource Institute for Healthy Utilization (BIHU), Zunyi Medical University, Zunyi, China.
Front Plant Sci. 2024 Jan 29;15:1302817. doi: 10.3389/fpls.2024.1302817. eCollection 2024.
Dendrobine, a valuable alkaloid found in , possesses significant pharmaceutical potential.
In this study, we explored innovative approaches to enhance dendrobine production by utilizing endophytic fungi in a Temporary Immersion Bioreactor System (TIBS, Nanjing BioFunction Co. Ltd., China) and traditional test bottles. Dendrobine was unequivocally identified and characterised in co-culture seedlings through UHPLC analysis and LC-MS qTOF analysis, supported by reference standards.
The CGTB (control group) and EGTB (experimental group) 12-month-old seedlings exhibited similar peak retention times at 7.6±0.1 minutes, with dendrobine identified as CHNO (molecular weight 264.195). The EGTB, co-cultured with (MD33), displayed a 2.6-fold dendrobine increase (1804.23 ng/ml) compared to the CGTB (685.95 ng/ml). Furthermore, a bioanalytical approach was applied to investigate the mono-culture of MD33 with or without seedlings in test bottles. The newly developed UHPLC-MS method allowed for dendrobine identification at a retention time of 7.6±0.1 minutes for control and 7.6±0.1 minutes for co-culture. Additionally, we explored TIBS to enhance dendrobine production. Co-culturing seedlings with (MD33) in the TIBS system led to a substantial 9.7-fold dendrobine increase (4415.77 ng/ml) compared to the control (454.01 ng/ml) after just 7 days. The comparative analysis of dendrobine concentration between EGTB and EGTIBS highlighted the remarkable potential of TIBS for optimizing dendrobine production. Future research may focus on scaling up the TIBS approach for commercial dendrobine production and investigating the underlying mechanisms for enhanced dendrobine biosynthesis in . The structural elucidation of dendrobine was achieved through H and C NMR spectroscopy, revealing a complex array of proton environments and distinct carbon environments, providing essential insights for the comprehensive characterization of the compound.
These findings hold promise for pharmaceutical and industrial applications of dendrobine and underline the role of endophytic fungi in enhancing secondary metabolite production in medicinal plants.
石斛碱是一种存在于[具体植物名称未给出]中的珍贵生物碱，具有巨大的药用潜力。
在本研究中，我们探索了创新方法，通过在临时浸没生物反应器系统（TIBS，南京博福生物科技有限公司，中国）和传统试验瓶中利用内生真菌来提高石斛碱产量。通过超高效液相色谱（UHPLC）分析和液相色谱 - 质谱联用四极杆飞行时间质谱（LC - MS qTOF）分析，并借助参考标准品，在共培养幼苗中明确鉴定并表征了石斛碱。
CGTB（对照组）和EGTB（实验组）12个月大的[植物名称未给出]幼苗在7.6±0.1分钟处显示出相似的峰保留时间，石斛碱被鉴定为C₁₅H₂₁NO₂（分子量264.195）。与[真菌名称未给出]（MD33）共培养的EGTB与CGTB（685.95纳克/毫升）相比，石斛碱含量增加了2.6倍（1804.23纳克/毫升）。此外，采用生物分析方法研究了在试验瓶中[真菌名称未给出]MD33与有无[植物名称未给出]幼苗的单培养情况。新开发的UHPLC - MS方法能够在保留时间7.6±0.1分钟（对照组）和7.6±0.1分钟（共培养组）鉴定出石斛碱。另外，我们探索了TIBS以提高石斛碱产量。在TIBS系统中将[植物名称未给出]幼苗与[真菌名称未给出]（MD33）共培养，仅7天后，与对照组（454.01纳克/毫升）相比，石斛碱含量大幅增加了9.7倍（4415.77纳克/毫升）。EGTB和EGTIBS之间石斛碱浓度的比较分析突出了TIBS在优化石斛碱生产方面的巨大潜力。未来的研究可能集中在扩大TIBS方法以实现商业化石斛碱生产，以及研究[具体植物名称未给出]中石斛碱生物合成增强的潜在机制。通过氢核磁共振（¹H NMR）和碳核磁共振（¹³C NMR）光谱实现了石斛碱的结构解析，揭示了一系列复杂的质子环境和不同的碳环境，为该化合物的全面表征提供了重要见解。
这些发现为石斛碱的制药和工业应用带来了希望，并强调了内生真菌在提高药用植物次生代谢产物产量中的作用。
