Forschungsbericht 2017



Development of New Methods in Fast Sampling and in Sample Processing for Microbial Metabolomics

Institut: V-1
Projektleitung: An-Ping Zeng
Stellvertretende Projektleitung: Philipp Arbter
Mitarbeiter/innen: Philipp Arbter
Lin Chen
Julian da Luz
Enrico Hans
Jan Sens
Laufzeit: 01.07.2006 — 01.12.2019
Finanzierung:Technische Universität Hamburg-Harburg (TUHH)

A rapid sampling and the following sample processing are critical for a reliable metabolomic analysis. Due to the complicated procedure and the high quantity of parameters there are many possibilities for variations and errors. The effective and sufficient separation of the exo- and endometabolome is a specific challenge in this context. Our aim is to develop new technologies and methods to minimize the errors and to design new experiments to investigate and describe cellular metabolism and its regulation. A new fully automated rapid sampling unit with on-chip filtration has been successfully developed which greatly improves the accuracy of intracellular metabolites, especially to overcome the problem of metabolic leakage. The concept combines flexibility and accuracy. The rapid sampling unit is based on a modular system. A specific valve system assures definite sample volumes and efficient mixing of the sample with the quenching solution. A special on-line and fast filtration system was implemented which can handle high cell density culture (up to OD value near 100). This is the first fast sampling system which can work under such extrem conditions more relevant to industrial bioprocesses. The system has been use to study different metabolic regulation in microorganisms such as E. coli and Corynebacterium glutamicum. 

Stichworte

  • metabolomics
  • Rapid sampling unit

Publikationen

  • da Luz, Julian A., Hans, Enrico, Frank, Doinita, Zeng, An-Ping: Analysis of intracellular metabolites of Corynebacterium glutamicum at high cell density with automated sampling and filtration and assessment of engineered enzymes for effective L-lysine production. Engineering in Life Sciences, 17: S. 512-522, 2017. , DOI: 10.1002/elsc.201600163
  • Julian A. da Luz, Enrico Hans and An-Ping Zeng : Improvement of intracellular metabolite analysis of microorganisms by automated fast filtration and on-filter quenching. Eng. Life Sciences: S. 135-142, 2014.
  • Christin Groeger, Wei Wang, Wael Sabra, Tyll Utesch and An-Ping Zeng: Metabolic and proteomic analyses of product selectivity and redox regulation in Clostridium pasteurianum grown on glycerol under varied iron availability. Microb. Cell Fact., 16(1): S. 64, 2017.
  • Zhou, Libang; Zeng, An-Ping : Metabolic engineering of Corynebacterium glutamicum for increasing lysine production by using a lysine riboswitch. ACS Synthetic Biology, 2014 (accepted).
  • Chen L, Zeng A-P: Rational Design and Metabolic Analysis of Escherichia coli for Effective Production of L-Tryptophan at High Concentration. Appl Microbiol Biotechnol , 15(101): S. 559-568, 2017. , DOI: doi: 10.1007/s00253-016-7772-5
  • Chen, Zhen and Rappert, Sugima and Zeng, An-Ping: Rational Design of Allosteric Regulation of Homoserine Dehydrogenase by a Nonnatural Inhibitor L-Lysine. ACS Synthetic Biology, 4(2): S. 126--131, feb 2015. , DOI: 10.1021/sb400133g, http://pubs.acs.org/doi/abs/10.1021/sb400133g