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Production of a periplasmic trehalase in Gluconobacter oxydans and growth on trehalose

Paper ID Volume ID Publish Year Pages File Format Full-Text
23062 43415 2014 9 PDF Available
Title
Production of a periplasmic trehalase in Gluconobacter oxydans and growth on trehalose
Abstract

•Expression of periplasmic enzymes in Gluconobacter oxydans.•Rational design to allow G. oxydans growth on trehalose.•Trehalase was successfully transported into the periplasm.•TreA expressing G. oxydans produced mainly acetate and 5-ketogluconate.•First step to engineer G. oxydans to use cheap bio-feedstocks.

Gluconobacter strains are specialized in the incomplete oxidation of monosaccharides. In contrast, growth and product formation from disaccharides is either very low or impossible. A pathway that allows growth on trehalose was rationally designed to broaden the substrate range of Gluconobacter oxydans. Expression vectors containing different signal sequences and the gene encoding alkaline phosphatase, phoA, from Escherichia coli were constructed. The signal peptide that exhibited the strongest periplasmic PhoA activity was used to generate a G. oxydans strain able to utilize the model disaccharide trehalose as a carbon and energy source by expressing the periplasmic trehalase TreA from E. coli. The strain had a doubling time of 3.7 h and reached a final optical density of 1.7 when trehalose was used as a growth substrate. In comparison, the wild-type harboring the empty vector and the strain expressing treA without a signal sequence grew slowly to a final OD of only 0.15. The trehalose concentration in treA expressing cultures decreased continuously during the exponential growth phase indicating that the substrate was hydrolyzed to glucose by TreA. In contrast to the wild-type growing on glucose, the treA expression strain mainly formed acetate and 5-ketogluconate as end products rather than gluconate.

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Keywords
Acetic acid bacteria; Incomplete oxidation; Substrate spectrum; Genetic tool; Signal peptide
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Production of a periplasmic trehalase in Gluconobacter oxydans and growth on trehalose
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 189, 10 November 2014, Pages 27–35
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
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Full-text PDF Download
Online Support
Any Questions? feel free to contact us