Efficient Hydroxylation of 1,8-cineole with Monoterpenoid-resistant Recombinant Pseudomonas Putida GS1

Overview

Journal World J Microbiol Biotechnol

Publisher Springer

Specialties Biotechnology
Microbiology

Date 2016 Jun 6

PMID 27263007

Citations 3

Authors

Jia Mi

Hendrik Schewe

Markus Buchhaupt

Dirk Holtmann

Jens Schrader

Affiliations

Soon will be listed here.

Abstract

In this work, monoterpenoid hydroxylation with Pseudomonas putida GS1 and KT2440 were investigated as host strains, and the cytochrome P450 monooxygenase CYP176A1 (P450cin) and its native redox partner cindoxin (CinC) from Citrobacter braakii were introduced in P. putida to catalyze the stereoselective hydroxylation of 1,8-cineole to (1R)-6β-hydroxy-1,8-cineole. Growth experiments in the presence of 1,8-cineole confirmed pseudomonads' superior resilience compared to E. coli. Whole-cell P. putida harboring P450cin with and without CinC were capable of hydroxylating 1,8-cineole, whereas coexpression of CinC has been shown to accelerate this bioconversion. Under the same conditions, P. putida GS1 produced more than twice the amount of heterologous P450cin and bioconversion product than P. putida KT2440. A concentration of 1.1 ± 0.1 g/L (1R)-6β-hydroxy-1,8-cineole was obtained within 55 h in shake flasks and 13.3 ± 1.9 g/L in 89 h in a bioreactor, the latter of which corresponds to a yield YP/S of 79 %. To the authors' knowledge, this is the highest product titer for a P450 based whole-cell monoterpene oxyfunctionalization reported so far. These results show that solvent-tolerant P. putida GS1 can be used as a highly efficient recombinant whole-cell biocatalyst for a P450 monooxygenase-based valorization of monoterpenoids.

Citing Articles

Industrial biotechnology of Pseudomonas putida: advances and prospects.

Weimer A, Kohlstedt M, Volke D, Nikel P, Wittmann C Appl Microbiol Biotechnol. 2020; 104(18):7745-7766.

PMID: 32789744 PMC: 7447670. DOI: 10.1007/s00253-020-10811-9.

Investigation of monoterpenoid resistance mechanisms in Pseudomonas putida and their consequences for biotransformations.

Schempp F, Hofmann K, Mi J, Kirchner F, Meffert A, Schewe H Appl Microbiol Biotechnol. 2020; 104(12):5519-5533.

PMID: 32296906 PMC: 7275096. DOI: 10.1007/s00253-020-10566-3.

Identifying and engineering the ideal microbial terpenoid production host.

Moser S, Pichler H Appl Microbiol Biotechnol. 2019; 103(14):5501-5516.

PMID: 31129740 PMC: 6597603. DOI: 10.1007/s00253-019-09892-y.

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