Rho D, Chauret N, Laberge N, Archambault J
Biotechnology Research Institute, National Research Council Canada, Montréal, Qc.
Appl Microbiol Biotechnol. 1992 Feb;36(5):611-7. doi: 10.1007/BF00183237.
Sanguinaria canadensis L. plants were harvested from a local forest and calli were initiated from leaf explants. The production of benzophenanthridine alkaloids (i.e. sanguinarine, sanguilutine, sanguirubine, chelerythrine, chelilutine and chelirubine) by S. canadensis cell grown in modified B5 and IM2 media was compared to the alkaloid content of rhizomes. Sanguinarine accounted for approximately 80% of the total alkaloid content of cultured cells (1.3%, g g-1) while sanguinarine and sanguirubine accounted for 70% of rhizome alkaloids (9.0%, g g-1). Sanguinarine, chelirubine and chelerythrine were the only known alkaloids detected in cultured S. canadensis cells. Maximum alkaloid production of cultures performed using B5 medium, containing half the original nitrate concentration, was observed following extracellular nitrate and sugar depletion. The scale-up of this culture was successfully performed in a 2-1 immobilization bioreactor. The consumption of sugar and nitrate as well as the oxygen (OTR) and carbon dioxide (CTR) transfer rates of the immobilized cell culture were monitored for 15 days. The maximum sugar and nitrate consumption rates were 1.8 g l-1 per day and 2.3 mM per day respectively. The maximum OTR and CTR of the immobilized cell culture were 0.8 mmol O2 l-1 h-1 and 0.95 mmol CO2 l-1 h-1 respectively. The sanguinarine yield of this culture reached 1.0% based on biomass dry weight (g g-1 dw) by day 15.
从当地森林采集加拿大血根草植株，并从叶片外植体诱导愈伤组织。将在改良B5和IM2培养基中生长的加拿大血根草细胞产生的苯并菲啶生物碱（即血根碱、血根黄素、血根红碱、白屈菜红碱、白屈菜黄素和白屈菜红碱）与根茎中的生物碱含量进行比较。血根碱约占培养细胞总生物碱含量的80%（1.3%，g g-1），而血根碱和血根红碱占根茎生物碱的70%（9.0%，g g-1）。血根碱、白屈菜红碱和白屈菜红碱是在培养的加拿大血根草细胞中检测到的仅有的已知生物碱。在细胞外硝酸盐和糖耗尽后，观察到使用硝酸盐浓度减半的B5培养基进行的培养物中生物碱产量最高。该培养物的放大培养在2-L固定化生物反应器中成功进行。对固定化细胞培养物的糖和硝酸盐消耗以及氧气（OTR）和二氧化碳（CTR）传递速率进行了15天的监测。糖和硝酸盐的最大消耗速率分别为每天1.8 g l-1和每天2.3 mM。固定化细胞培养物的最大OTR和CTR分别为0.8 mmol O2 l-1 h-1和0.95 mmol CO2 l-1 h-1。到第15天，该培养物的血根碱产量基于生物量干重达到1.0%（g g-1 dw）。
