Resolution of alkaloid racemate: a novel microbial approach for the production of enantiopure lupanine via industrial wastewater valorization.

Ecotoxicological assessment Enantiomeric excess Enantiomers Gene expression Lupanine Pseudomonas putida LPK411 Quantitative real-time PCR

Journal

Microbial cell factories
ISSN: 1475-2859
Titre abrégé: Microb Cell Fact
Pays: England
ID NLM: 101139812

Informations de publication

Date de publication:
14 Mar 2020
Historique:
received: 25 12 2019
accepted: 04 03 2020
entrez: 15 3 2020
pubmed: 15 3 2020
medline: 22 10 2020
Statut: epublish

Résumé

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.

Sections du résumé

BACKGROUND BACKGROUND
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.
RESULTS RESULTS
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.
CONCLUSIONS CONCLUSIONS
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.

Identifiants

pubmed: 32169079
doi: 10.1186/s12934-020-01324-1
pii: 10.1186/s12934-020-01324-1
pmc: PMC7071741
doi:

Substances chimiques

Waste Water 0
lupanine 183KU7535A
Sparteine 298897D62S

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

67

Subventions

Organisme : Cyprus Research and Innovation Foundation
ID : ΚΟΙΝΑ/ΠΚΠ-WATER/0315/14
Organisme : Fundação para a Ciência e a Tecnologia
ID : WaterJPI/0001/2014
Organisme : Fundação para a Ciência e a Tecnologia
ID : WaterJPI/0002/2014
Organisme : Fundação para a Ciência e a Tecnologia
ID : WaterJPI/0003/2014

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Auteurs

Stella Parmaki (S)

Department of Chemical Engineering, Cyprus University of Technology, 30 Archbishop Kyprianou Str., 3036, Limassol, Cyprus.

Argyro Tsipa (A)

Civil and Environmental Engineering, University of Cyprus, 75 Kallipoleos Str., 1678, Nicosia, Cyprus.

Marlen I Vasquez (MI)

Department of Chemical Engineering, Cyprus University of Technology, 30 Archbishop Kyprianou Str., 3036, Limassol, Cyprus.

João M J Gonçalves (JMJ)

Research Institute for Medicines (iMed. ULisboa), Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003, Lisbon, Portugal.

Ioanna Hadjiadamou (I)

Department of Chemistry, University of Cyprus, P.O. Box 20537, 1678, Nicosia, Cyprus.
Department of Agricultural Sciences, Biotechnology and Food Science, Cyprus University of Technology, 30 Archbishop Kyprianou Str., 3036, Limassol, Cyprus.

Frederico C Ferreira (FC)

Institute for Bioengineering and Biosciences, Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001, Lisbon, Portugal.

Carlos A M Afonso (CAM)

Research Institute for Medicines (iMed. ULisboa), Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003, Lisbon, Portugal.

Chrysoulla Drouza (C)

Department of Agricultural Sciences, Biotechnology and Food Science, Cyprus University of Technology, 30 Archbishop Kyprianou Str., 3036, Limassol, Cyprus.

Michalis Koutinas (M)

Department of Chemical Engineering, Cyprus University of Technology, 30 Archbishop Kyprianou Str., 3036, Limassol, Cyprus. michail.koutinas@cut.ac.cy.

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Classifications MeSH