Title: Production efficiency of the bacterial non-ribosomal peptide indigoidine relies on the respiratory metabolic state in S. cerevisiae

Abstract

Background: Beyond pathway engineering, the metabolic state of the production host is critical in maintaining the efficiency of cellular production. The biotechnologically important yeast Saccharomyces cerevisiae adjusts its energy metabolism based on the availability of oxygen and carbon sources. This transition between respiratory and non-respiratory metabolic state is accompanied by substantial modifications of central carbon metabolism, which impact the efficiency of metabolic pathways and the corresponding final product titers. Non-ribosomal peptide synthetases (NRPS) are an important class of biocatalysts that provide access to a wide array of secondary metabolites. Indigoidine, a blue pigment, is a representative NRP that is valuable by itself as a renewably produced pigment. Results: Saccharomyces cerevisiae was engineered to express a bacterial NRPS that converts glutamine to indigoidine. We characterize carbon source use and production dynamics, and demonstrate that indigoidine is solely produced during respiratory cell growth. Production of indigoidine is abolished during non-respiratory growth even under aerobic conditions. By promoting respiratory conditions via controlled feeding, we scaled the production to a 2 L bioreactor scale, reaching a maximum titer of 980 mg/L. In conclusion, this study represents the first use of the Streptomyces lavendulae NRPS (BpsA) in a fungal host and its scale-up. The finalmore » product indigoidine is linked to the activity of the TCA cycle and serves as a reporter for the respiratory state of S. cerevisiae. Our approach can be broadly applied to investigate diversion of flux from central carbon metabolism for NRPS and other heterologous pathway engineering, or to follow a population switch between respiratory and non-respiratory modes.« less

Authors:

Wehrs, Maren; Prahl, Jan-Philip; Moon, Jadie; Li, Yuchen; Tanjore, Deepti; Keasling, Jay D.; Pray, Todd; Mukhopadhyay, Aindrila

• Search DOE PAGES for author "Mukhopadhyay, Aindrila"
• Search DOE PAGES for ORCID "0000-0002-6513-7425"

Publication Date:

Thu Dec 13 00:00:00 EST 2018

Research Org.:

Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Org.:

USDOE Office of Science (SC), Biological and Environmental Research (BER)

OSTI Identifier:

1618894

Alternate Identifier(s):

OSTI ID: 1493277

Grant/Contract Number:  

AC02-05CH11231

Resource Type:

Published Article

Journal Name:

Microbial Cell Factories

Additional Journal Information:

Journal Name: Microbial Cell Factories Journal Volume: 17 Journal Issue: 1; Journal ID: ISSN 1475-2859

Publisher:

Springer Science + Business Media

Country of Publication:

United Kingdom

Language:

English

Subject:

59 BASIC BIOLOGICAL SCIENCES; Non-ribosomal peptide synthetase; NRPS, S. cerevisiae; Indigoidine; Metabolic state; TCA cycle activity; Non-ribosomal peptide synthesis; Bioreactor; BpsA; BJ5465