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Enhanced production of trehalose in Escherichia coli by homologous expression of otsBA in the presence of the trehalase inhibitor, validamycin A, at high osmolarity.
J Biosci Bioeng 2012; 113(2):224-32JB

Abstract

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.

Authors+Show Affiliations

Department of Chemical and Biological Engineering and ERI, GyeongSang National University, 900 Gajwadong, Jinju, Gyeongnam 660-701, Republic of Korea.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

22036231

Citation

Li, He, et al. "Enhanced Production of Trehalose in Escherichia Coli By Homologous Expression of otsBA in the Presence of the Trehalase Inhibitor, Validamycin A, at High Osmolarity." Journal of Bioscience and Bioengineering, vol. 113, no. 2, 2012, pp. 224-32.
Li H, Su H, Kim SB, et al. Enhanced production of trehalose in Escherichia coli by homologous expression of otsBA in the presence of the trehalase inhibitor, validamycin A, at high osmolarity. J Biosci Bioeng. 2012;113(2):224-32.
Li, H., Su, H., Kim, S. B., Chang, Y. K., Hong, S. K., Seo, Y. G., & Kim, C. J. (2012). Enhanced production of trehalose in Escherichia coli by homologous expression of otsBA in the presence of the trehalase inhibitor, validamycin A, at high osmolarity. Journal of Bioscience and Bioengineering, 113(2), pp. 224-32. doi:10.1016/j.jbiosc.2011.09.018.
Li H, et al. Enhanced Production of Trehalose in Escherichia Coli By Homologous Expression of otsBA in the Presence of the Trehalase Inhibitor, Validamycin A, at High Osmolarity. J Biosci Bioeng. 2012;113(2):224-32. PubMed PMID: 22036231.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Enhanced production of trehalose in Escherichia coli by homologous expression of otsBA in the presence of the trehalase inhibitor, validamycin A, at high osmolarity. AU - Li,He, AU - Su,Hong, AU - Kim,Sung Bae, AU - Chang,Yong Keun, AU - Hong,Soon-Kwang, AU - Seo,Yang-Gon, AU - Kim,Chang-Joon, Y1 - 2011/10/26/ PY - 2011/05/03/received PY - 2011/09/20/revised PY - 2011/09/24/accepted PY - 2011/11/1/entrez PY - 2011/11/1/pubmed PY - 2012/9/29/medline SP - 224 EP - 32 JF - Journal of bioscience and bioengineering JO - J. Biosci. Bioeng. VL - 113 IS - 2 N2 - 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. SN - 1347-4421 UR - https://www.unboundmedicine.com/medline/citation/22036231/Enhanced_production_of_trehalose_in_Escherichia_coli_by_homologous_expression_of_otsBA_in_the_presence_of_the_trehalase_inhibitor_validamycin_A_at_high_osmolarity_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1389-1723(11)00391-4 DB - PRIME DP - Unbound Medicine ER -