Enhanced production of trehalose in Escherichia coli by homologous expression of otsBA in the presence of the trehalase inhibitor, validamycin A, at high osmolarity

Li, HeDepartment of Chemical and Biological Engineering and ERI, GyeongSang National University, 900 Gajwadong, Jinju, Gyeongnam 660-701, Republic of Korea (författare)

Trehalose production in Escherichia coli DH5α was explored by overexpressing otsBA operon encoding trehalose-6-phosphate synthase and trehalose-6-phosphate phosphatase. Production and subsequent degradation of trehalose resulted in low production of trehalose in engineered cells overexpressing otsBA, which was primarily due to the concomitant expression of endogenous trehalase. Through an in vitro enzyme assay and flask cultures of engineered cells, trehalase expression was shown to be directly related to the expression of otsBA rather than osmotic stress. Validamycin A effectively inhibited E. coli trehalase and the intracellular accumulation of trehalose was markedly enhanced in the presence of validamycin A at an optimal concentration in the medium. The trehalose production was further increased upon growth in a hypertonic medium in the presence of validamycin A, with most trehalose accumulating as an intracellular product. The highest titer was obtained when otsBA expression was induced by a medium-copy vector, ptrc99A, with 0.5mM of isopropyl β-d-1-thiogalactopyranoside. Trehalose titer was 1.7 g/L in controlled bioreactor cultures using synthetic M9 medium supplemented with 40 g/L glycerol, 0.1mM validamycin A, and 300 mM NaCl.

Su, HongDepartment of Chemical and Biological Engineering and ERI, GyeongSang National University, 900 Gajwadong, Jinju, Gyeongnam 660-701, Republic of Korea (författare)
Kim, Sung BaeDepartment of Chemical and Biological Engineering and ERI, GyeongSang National University, 900 Gajwadong, Jinju, Gyeongnam 660-701, Republic of Korea (författare)
Chang, Yong KeunDepartment of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology, 373-1 Gusung-dong, Yuseong-gu, Daejeon 305-701, Republic of Korea (författare)
Hong, Soon-KwangDivision of Bioscience and Bioinformatics, Myongji University, Yongin 449-728, Republic of Korea (författare)
Seo, Yang-GonDepartment of Chemical and Biological Engineering and ERI, GyeongSang National University, 900 Gajwadong, Jinju, Gyeongnam 660-701, Republic of Korea (författare)
Kim, Chang-JoonDepartment of Chemical and Biological Engineering and ERI, GyeongSang National University, 900 Gajwadong, Jinju, Gyeongnam 660-701, Republic of Korea (författare)
Department of Chemical and Biological Engineering and ERI, GyeongSang National University, 900 Gajwadong, Jinju, Gyeongnam 660-701, Republic of KoreaDepartment of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology, 373-1 Gusung-dong, Yuseong-gu, Daejeon 305-701, Republic of Korea (creator_code:org_t)

Ingår i:Journal of Bioscience and Bioengineering: Elsevier113:2, s. 224-2321389-17231347-4421

Av författaren/redakt...: Li, He; Su, Hong; Kim, Sung Bae; Chang, Yong Keun; Hong, Soon-Kwang; Seo, Yang-Gon; visa fler...; Kim, Chang-Joon; visa färre...

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