Universität zu Köln

Roenneke, Benjamin, Rosenfeldt, Natalie, Derya, Sami M., Novak, Jens F., Marin, Kay, Kraemer, Reinhard and Seibold, Gerd M. (2018). Production of the compatible solute alpha-D-glucosylglycerol by metabolically engineered Corynebacterium glutamicum. Microb. Cell. Fact., 17. LONDON: BIOMED CENTRAL LTD. ISSN 1475-2859

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Abstract

Background: alpha-D-Glucosylglycerol (alpha GG) has beneficial functions as a moisturizing agent in cosmetics and potential as a health food material, and therapeutic agent. alpha GG serves as compatible solute in various halotolerant cyanobacteria such as Synechocystis sp. PCC 6803, which synthesizes alpha GG in a two-step reaction: The enzymatic condensation of ADP-glucose and glycerol 3-phosphate by GG-phosphate synthase (GGPS) is followed by the dephosphorylation of the intermediate by the GG-phosphate phosphatase (GGPP). The Gram-positive Corynebacterium glutamicum, an industrial workhorse for amino acid production, does not utilize alpha GG as a substrate and was therefore chosen for the development of a heterologous microbial production platform for alpha GG. Results: Plasmid-bound expression of ggpS and ggpP from Synechocystis sp. PCC 6803 enabled alpha GG synthesis exclusively in osmotically stressed cells of C. glutamicum (pEKEx2-ggpSP), which is probably due to the unique intrinsic control mechanism of GGPS activity in response to intracellular ion concentrations. C. glutamicum was then engineered to optimize precursor supply for alpha GG production: The precursor for alpha GG synthesis ADP-glucose gets metabolized by both the glgA encoded glycogen synthase and the otsA encoded trehalose-6-phosphate synthase. Upon deletion of both genes the alpha GG concentration in culture supernatants was increased from 0.5 mM in C. glutamicum (pEKEx3-ggpSP) to 2.9 mM in C. glutamicum Delta otsA IMglgA (pEKEx3-ggpSP). Upon nitrogen limitation, which inhibits synthesis of amino acids as compatible solutes, C. glutamicum Delta otsA IMglgA (pEKEx3-ggpSP) produced more than 10 mM alpha GG (about 2 g L-1). Conclusions: Corynebacterium glutamicum can be engineered as efficient platform for the production of the compatible solute alpha GG. Redirection of carbon flux towards alpha GG synthesis by elimination of the competing pathways for glycogen and trehalose synthesis as well as optimization of nitrogen supply is an efficient strategy to further optimize production of alpha GG.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Roenneke, Benjamin UNSPECIFIED UNSPECIFIED UNSPECIFIED Rosenfeldt, Natalie UNSPECIFIED UNSPECIFIED UNSPECIFIED Derya, Sami M. UNSPECIFIED UNSPECIFIED UNSPECIFIED Novak, Jens F. UNSPECIFIED UNSPECIFIED UNSPECIFIED Marin, Kay UNSPECIFIED UNSPECIFIED UNSPECIFIED Kraemer, Reinhard UNSPECIFIED UNSPECIFIED UNSPECIFIED Seibold, Gerd M. UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-182805
DOI:	10.1186/s12934-018-0939-2
Journal or Publication Title:	Microb. Cell. Fact.
Volume:	17
Date:	2018
Publisher:	BIOMED CENTRAL LTD
Place of Publication:	LONDON
ISSN:	1475-2859
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language ADP-GLUCOSE PYROPHOSPHORYLASE; BACTERIAL GLYCOGEN-SYNTHESIS; SYNECHOCYSTIS SP PCC-6803; ESCHERICHIA-COLI; TREHALOSE METABOLISM; OSMOTIC-STRESS; MECHANOSENSITIVE CHANNELS; HETEROLOGOUS EXPRESSION; LIPID-COMPOSITION; GLGX GENE Multiple languages Biotechnology & Applied Microbiology Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/18280