1. G6P-capturing molecules in the periplasm of Escherichia coli accelerate the shikimate pathway.
- Author
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Fujiwara, Ryosuke, Nakano, Mariko, Hirata, Yuuki, Otomo, Chisako, Nonaka, Daisuke, Kawada, Sakiya, Nakazawa, Hikaru, Umetsu, Mitsuo, Shirai, Tomokazu, Noda, Shuhei, Tanaka, Tsutomu, and Kondo, Akihiko
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PERIPLASM , *CARRIER proteins , *IMMOBILIZED proteins , *SMALL molecules , *GLUCOSE transporters , *GLUCOSIDASES , *CHEMICAL synthesis - Abstract
Escherichia coli , the most studied prokaryote, is an excellent host for producing valuable chemicals from renewable resources as it is easy to manipulate genetically. Since the periplasmic environment can be easily controlled externally, elucidating how the localization of specific proteins or small molecules in the periplasm affects metabolism may lead to bioproduction development using E. coli. We investigated metabolic changes and its mechanisms occurring when specific proteins are localized to the E. coli periplasm. We found that the periplasmic localization of β-glucosidase promoted the shikimate pathway involved in the synthesis of aromatic chemicals. The periplasmic localization of other proteins with an affinity for glucose-6-phosphate (G6P), such as inactivated mutants of Pgi, Zwf, and PhoA, similarly accelerated the shikimate pathway. Our results indicate that G6P is transported from the cytoplasm to the periplasm by the glucose transporter protein EIICBGlc, and then captured by β-glucosidase. • Periplasmic expression of β-glucosidase (BGL) increases L-Phe production in E. coli. • Two key factors involved in this phenomenon, G6P and EIICGlc domain, were defined. • Cytoplasmic G6P is secreted into the periplasm via EIICGlc domain and captured by BGL. • Periplasmic expression of other G6P-capturing proteins also increase L-Phe production. • This technique can be applied to produce other shikimate pathway derivatives. [ABSTRACT FROM AUTHOR]
- Published
- 2022
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