Arano-Varela H, Fernández F J, Estrada-Zúñiga M E, Cruz-Sosa F
1Departamento de Biotecnología, Universidad Autónoma Metropolitana-Iztapalapa, San Rafael Atlixco No. 186, Col. Vicentina, CP 09340 Mexico City, Mexico.
2Centro de Investigación en Recursos Bióticos-Facultad de Ciencias, Universidad Autónoma del Estado de Mexico, Carretera Toluca-Ixtlahuaca km 14.5, C.P. 50285 Toluca, Estado de Mexico Mexico.
3 Biotech. 2020 Jun;10(6):245. doi: 10.1007/s13205-020-02222-4. Epub 2020 May 11.
Previously, our group reported the establishment of a white callus cell line of Kunth that is a high producer of the secondary metabolite, verbascoside (VB, also named acteoside), under suspension culture conditions. Here, we present experimental evidence of the sustained ability of that cellular line to grow and produce high amounts of VB for 5 years of continuous culture. Cellular line profiles were determined at the early (at the beginning) and late stages (at the end of 5 years of continuous subculturing) by analyzing relevant parameters of culture growth, i.e., specific growth rate [], doubling time [dt], and growth index [GI], as well as VB production. Late-stage cultures exhibited a 61% faster growth rate than early-stage subcultures, and 25 and 3% lower doubling time and growth index. The extents of growth phases were found to be different. Similar amounts of biomass were found (9.5 g and 9.4 g L). Verbascoside production increased parallel to cell growth; maximal yield level occurred in the mid-exponential phase and lasted until the end of the stationary phase (i.e., from the 15th to the 25th day and from the 9th to the 21st day for the early and late stages, correspondingly). The content of VB was higher in the late-stage culture (1.43 ± 0945 g L) than in the early-stage culture (1.21 ± 0.0286 g L). Productivity values point out the potential use of . cell line in the biotechnological production of VB and for research focused on the biochemistry of secondary metabolism.
此前,我们团队报道了建立了一种昆特白愈伤组织细胞系,该细胞系在悬浮培养条件下是次生代谢产物毛蕊花糖苷(VB,也称为洋丁香酚苷)的高产生产者。在此,我们提供了实验证据,证明该细胞系在连续培养5年中具有持续生长和大量生产VB的能力。通过分析培养生长的相关参数,即比生长速率[μ]、倍增时间[dt]和生长指数[GI]以及VB产量,在早期(开始时)和后期(连续继代培养5年后)确定细胞系概况。后期培养物的生长速率比早期继代培养物快61%,倍增时间和生长指数分别低25%和3%。发现生长阶段的程度不同。发现生物量的量相似(9.5 g和9.4 g L)。毛蕊花糖苷的产量与细胞生长平行增加;最大产量水平出现在指数中期,并持续到稳定期结束(即早期和后期分别从第15天到第25天和从第9天到第21天)。后期培养物中VB的含量(1.43±0.0945 g L)高于早期培养物(1.21±0.0286 g L)。生产力值指出了该细胞系在VB生物技术生产以及专注于次生代谢生物化学研究中的潜在用途。