Lupanine is a plant toxin contained in the wastewater of lupine bean processing industries, which could be used for semi-synthesis of various novel high added-value compounds. This paper introduces an environmental friendly process for microbial production of enantiopure lupanine. Previously isolated P. putida LPK411, R. rhodochrous LPK211 and Rhodococcus sp. LPK311, holding the capacity to utilize lupanine as single carbon source, were employed as biocatalysts for resolution of racemic lupanine. All strains achieved high enantiomeric excess (ee) of L-(−)-lupanine (> 95%), while with the use of LPK411 53% of the initial racemate content was not removed. LPK411 fed with lupanine enantiomers as single substrates achieved 92% of D-(+)-lupanine biodegradation, whereas L-(−)-lupanine was not metabolized. Monitoring the transcriptional kinetics of the luh gene in cultures supplemented with the racemate as well as each of the enantiomers supported the enantioselectivity of LPK411 for D-(+)-lupanine biotransformation, while (trans)-6-oxooctahydro-1H-quinolizine-3-carboxylic acid was detected as final biodegradation product from D-(+)-lupanine use. Ecotoxicological assessment demonstrated that lupanine enantiomers were less toxic to A. fischeri compared to the racemate exhibiting synergistic interaction. The biological chiral separation process of lupanine presented here constitutes an eco-friendly and low-cost alternative to widely used chemical methods for chiral separation.

中文翻译：

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生物碱外消旋体的拆分：通过工业废水增值生产对映体纯的Lupanine的新型微生物方法

Lupanine是羽扇豆加工工业废水中所含的一种植物毒素，可用于半合成各种新型高附加值化合物。本文介绍了一种环境友好的工艺生产对映体纯的Lupanine。先前分离的恶臭假单胞菌LPK411，杜鹃红球菌LPK211和红球菌sp。LPK311，具有利用Lupanine作为单一碳源的能力，被用作拆分外消旋Lupanine的生物催化剂。所有菌株均实现了L-（-）-鲁潘宁（> 95％）的高对映体过量（ee），而使用LPK411则未去除53％的初始外消旋物含量。饲喂Lupanine对映体作为单一底物的LPK411实现了92％的D-（+）-Lupanine生物降解，而L-（-）-Lupanine不被代谢。监测外消旋物以及每种对映异构体补充的培养物中luh基因的转录动力学支持LPK411对D-（+）-Lupanine生物转化的对映选择性，而（trans）-6-氧代八氢-1H-quinolizine-3从D-（+）-鲁潘宁的使用中检测到-羧酸是最终的生物降解产物。生态毒理学评估表明，与表现出协同作用的外消旋体相比，Lupanine对映体对费氏曲霉的毒性较小。此处介绍的Lupanine的生物手性分离过程构成了一种广泛使用的手性分离化学方法的环保且低成本的替代品。而从D-（+）-Lupanine的使用中检测到（反式）-6-氧代八氢-1H-喹啉-3羧酸是最终的生物降解产物。生态毒理学评估表明，与表现出协同作用的外消旋体相比，Lupanine对映体对费氏曲霉的毒性较小。此处介绍的Lupanine的生物手性分离过程构成了一种广泛使用的手性分离化学方法的环保且低成本的替代品。而从D-（+）-Lupanine的使用中检测到（反式）-6-氧代八氢-1H-喹啉-3羧酸是最终的生物降解产物。生态毒理学评估表明，与表现出协同作用的外消旋体相比，Lupanine对映体对费氏曲霉的毒性较小。此处介绍的Lupanine的生物手性分离过程构成了一种广泛使用的手性分离化学方法的环保且低成本的替代品。