Production of tetra-methylpyrazine using engineered Corynebacterium glutamicum

Thomas Eng; Yusuke Sasaki; Robin A Herbert; Andrew Lau; Jessica Trinh; Yan Chen; Mona Mirsiaghi; Christopher J Petzold; Aindrila Mukhopadhyay

doi:10.1016/j.mec.2019.e00115

Production of tetra-methylpyrazine using engineered Corynebacterium glutamicum

Metab Eng Commun. 2019 Dec 2:10:e00115. doi: 10.1016/j.mec.2019.e00115. eCollection 2020 Jun.

Authors

Thomas Eng^{1

2}, Yusuke Sasaki^{1

2

3

4}, Robin A Herbert^{1

2}, Andrew Lau^{1

2}, Jessica Trinh^{1

2}, Yan Chen^{1

2}, Mona Mirsiaghi^{2

5}, Christopher J Petzold^{1

2}, Aindrila Mukhopadhyay^{1

2

6}

Affiliations

¹ Joint BioEnergy Institute, Emeryville, CA, USA.
² Biological Systems and Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
³ Graduate School of Advanced Integrated Studies in Human Survivability, Kyoto University, Sakyo-ku, Kyoto, Japan.
⁴ Japan Society for the Promotion of Science, Sakyo-ku, Kyoto, Japan.
⁵ Advanced Biofuels Process Demonstration Unit, Lawrence Berkeley National Laboratory, Emeryville, CA, USA.
⁶ Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.

Abstract

Corynebacterium glutamicum ATCC 13032 is an established and industrially-relevant microbial host that has been utilized for the expression of many desirable bioproducts. Tetra-methylpyrazine (TMP) is a naturally occurring alkylpyrazine with broad applications spanning fragrances to resins. We identified an engineered strain of C. glutamicum which produces 5 g/L TMP and separately, a strain which can co-produce both TMP and the biofuel compound isopentenol. Ionic liquids also stimulate TMP production in engineered strains. Using a fed batch-mode feeding strategy, ionic liquid stimulated strains produced 2.2 g/L of tetra-methylpyrazine. We show that feedback from a specific heterologous gene pathway on host physiology leads to acetoin accumulation and the production of TMP.

Keywords: 2,3,5,6-Tetra-methylpyrazine; Alkaloids; Bioreactor; Corynebacterium glutamicum; Isopentenol; Terpene.