title: Photosynthetic production of enantioselective biocatalysts.
abstract: BACKGROUND
  Global resource depletion poses a dramatic threat to our society and creates a strong demand for alternative resources that do not compete with the production of food. Meeting this challenge requires a thorough rethinking of all steps of the value chain regarding their sustainability resource demand and the possibility to substitute current, petrol-based supply-chains with renewable resources. This regards also the production of catalysts for chemical synthesis. Phototrophic microorganisms have attracted considerable attention as a biomanufacturing platform for the sustainable production of chemicals and biofuels. They allow the direct utilization of carbon dioxide and do not compete with food production. Photosynthetic enzyme production of catalysts would be a sustainable supply of these important components of the biotechnological and chemical industries. This paper focuses on the usefulness of recombinant cyanobacteria for the photosynthetic expression of enantioselective catalysts. As a proof of concept, we used the cyanobacterium Synechocystis sp. PCC 6803 for the heterologous expression of two highly enantioselective enzymes.
  
  
  RESULTS
  We investigated the expression yield and the usefulness of cyanobacterial cell extracts for conducting stereoselective reactions. The cyanobacterial enzyme expression achieved protein yields of 3% of total soluble protein (%TSP) while the expression in E. coli yielded 6-8% TSP. Cell-free extracts from a recombinant strain expressing the recombinant esterase ST0071 from the thermophilic organism Sulfolobus tokodai ST0071 and arylmalonate decarboxylase from Bordetella bronchiseptica showed excellent enantioselectivity (>99% ee) and yield (>91%) in the desymmetrisation of prochiral malonates.
  
  
  CONCLUSIONS
  We were able to present the proof-of-concept of photoautotrophic enzyme expression as a viable alternative to heterotrophic expression hosts. Our results show that the introduction of foreign genes is straightforward. Cell components from Synechocystis did not interfere with the stereoselective transformations, underlining the usability of photoautotrophic organisms for the production of enzymes. Given the considerable commercial value of recombinant biocatalysts, cyanobacterial enzyme expression has thus the potential to complement existing approaches to use phototrophic organisms for the production of chemicals and biofuels.
authors.affiliation: Junior Research Group for Microbial Biotechnology, Ruhr-Universität Bochum, Universitätsstr. 150, 44780, Bochum, Germany. Maik.Bartsch@rub.de.
authors.forename: Maik
authors.initials: M
authors.lastname: Bartsch
authors.affiliation: Junior Research Group for Microbial Biotechnology, Ruhr-Universität Bochum, Universitätsstr. 150, 44780, Bochum, Germany. Sarah.Gassmeyer@rub.de.
authors.forename: Sarah K
authors.initials: SK
authors.lastname: Gassmeyer
authors.affiliation: Junior Research Group for Microbial Biotechnology, Ruhr-Universität Bochum, Universitätsstr. 150, 44780, Bochum, Germany. Katharina.Koeninger@rub.de.
authors.forename: Katharina
authors.initials: K
authors.lastname: Köninger
authors.affiliation: Department of Biosciences and Informatics, Keio University, 3-14-1 Hiyoshi, Yokohama, 223-8522, Japan. K.Igarashi.0815@gmail.com.
authors.forename: Kosuke
authors.initials: K
authors.lastname: Igarashi
authors.affiliation: Chair for Plant Biochemistry, Ruhr-Universität Bochum, Universitätsstr. 150, 44780, Bochum, Germany. Pasqual.Liauw@rub.de.
authors.forename: Pasqual
authors.initials: P
authors.lastname: Liauw
authors.affiliation: Chair for Plant Biochemistry, Ruhr-Universität Bochum, Universitätsstr. 150, 44780, Bochum, Germany. Nina.G.Dyczm@rub.de.
authors.forename: Nina
authors.initials: N
authors.lastname: Dyczmons-Nowaczyk
authors.affiliation: Department of Biosciences and Informatics, Keio University, 3-14-1 Hiyoshi, Yokohama, 223-8522, Japan. kmiyamoto@bio.keio.ac.jp.
authors.forename: Kenji
authors.initials: K
authors.lastname: Miyamoto
authors.affiliation: Department of Biosciences and Informatics, Keio University, 3-14-1 Hiyoshi, Yokohama, 223-8522, Japan. Marc.M.Nowaczyk@rub.de.
authors.forename: Marc M
authors.initials: MM
authors.lastname: Nowaczyk
authors.affiliation: Junior Research Group for Microbial Biotechnology, Ruhr-Universität Bochum, Universitätsstr. 150, 44780, Bochum, Germany. Robert.Kourist@rub.de.
authors.forename: Robert
authors.initials: R
authors.lastname: Kourist
mesh_terms: D019843:Archaeal Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D002384:Catalysis; D004798:Enzymes / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D004950:Esterases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D010788:Photosynthesis; D016944:Sulfolobus / Q000201:enzymology*; D046939:Synechocystis / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism
chemical_list: D019843:Archaeal Proteins; D004798:Enzymes; D004950:Esterases