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Title: Redirection of the Glycolytic Flux Enhances Isoprenoid Production in Saccharomyces cerevisiae

Abstract

Sufficient supply of reduced nicotinamide adenine dinucleotide phosphate (NADPH) is a prerequisite of the overproduction of isoprenoids and related bioproducts in Saccharomyces cerevisiae. Although S. cerevisiae highly depends on the oxidative pentose phosphate (PP) pathway to produce NADPH, its metabolic flux toward the oxidative PP pathway is limited due to the rigid glycolysis flux. To maximize NADPH supply for the isoprenoid production in yeast, upper glycolytic metabolic fluxes are reduced by introducing mutations into phosphofructokinase (PFK) along with overexpression of ZWF1 encoding glucose-6-phosphate (G6P) dehydrogenase. The PFK mutations (Pfk1 S724D and Pfk2 S718D) result in less glycerol production and more accumulation of G6P, which is a gateway metabolite toward the oxidative PP pathway. When combined with the PFK mutations, overexpression of ZWF1 caused substantial increases of [NADPH]/[NADP+] ratios whereas the effect of ZWF1 overexpression alone in the wild-type strain is not noticeable. Also, the introduction of ZWF1 overexpression and the PFK mutations into engineered yeast overexpressing acetyl-CoA C-acetyltransferase (ERG10), truncated HMG-CoA reductase isozyme 1 (tHMG1), and amorphadiene synthase (ADS) leads to a titer of 497 mg L–1 of amorphadiene (3.7-fold over the parental strain). These results suggest that perturbation of upper glycolytic fluxes, in addition to ZWF1 overexpression, is necessarymore » for efficient NADPH supply through the oxidative PP pathway and enhanced production of isoprenoids by engineered S. cerevisiae.« less

Authors:
ORCiD logo [1];  [2];  [1];  [1];  [3]; ORCiD logo [1]
  1. Univ. of Illinois, Urbana-Champaign, IL (United States). Dept. of Food Science and Human Nutrition and Carl R. Woese Institute for Genomic Biology
  2. Univ. of Illinois, Urbana-Champaign, IL (United States). Carl R. Woese Institute for Genomic Biology; Korea Univ., Seoul (Korea). Dept. of Biotechnology
  3. Korea Univ., Seoul (Korea). Dept. of Biotechnology
Publication Date:
Research Org.:
Center for Advanced Bioenergy and Bioproducts Innovation (CABBI)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER)
OSTI Identifier:
1559936
Alternate Identifier(s):
OSTI ID: 1562316
Grant/Contract Number:  
SC0018420; DE‐SC0018420
Resource Type:
Accepted Manuscript
Journal Name:
Biotechnology Journal
Additional Journal Information:
Journal Volume: none; Journal Issue: none; Journal ID: ISSN 1860-6768
Publisher:
Wiley
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; isoprenoid, NADPH, oxidative pentose phosphate pathway, phosphofructokinase, Saccharomyces cerevisiae

Citation Formats

Kwak, Suryang, Yun, Eun Ju, Lane, Stephan, Oh, Eun Joong, Kim, Kyoung Heon, and Jin, Yong‐Su. Redirection of the Glycolytic Flux Enhances Isoprenoid Production in Saccharomyces cerevisiae. United States: N. p., 2019. Web. doi:10.1002/biot.201900173.
Kwak, Suryang, Yun, Eun Ju, Lane, Stephan, Oh, Eun Joong, Kim, Kyoung Heon, & Jin, Yong‐Su. Redirection of the Glycolytic Flux Enhances Isoprenoid Production in Saccharomyces cerevisiae. United States. doi:10.1002/biot.201900173.
Kwak, Suryang, Yun, Eun Ju, Lane, Stephan, Oh, Eun Joong, Kim, Kyoung Heon, and Jin, Yong‐Su. Thu . "Redirection of the Glycolytic Flux Enhances Isoprenoid Production in Saccharomyces cerevisiae". United States. doi:10.1002/biot.201900173. https://www.osti.gov/servlets/purl/1559936.
@article{osti_1559936,
title = {Redirection of the Glycolytic Flux Enhances Isoprenoid Production in Saccharomyces cerevisiae},
author = {Kwak, Suryang and Yun, Eun Ju and Lane, Stephan and Oh, Eun Joong and Kim, Kyoung Heon and Jin, Yong‐Su},
abstractNote = {Sufficient supply of reduced nicotinamide adenine dinucleotide phosphate (NADPH) is a prerequisite of the overproduction of isoprenoids and related bioproducts in Saccharomyces cerevisiae. Although S. cerevisiae highly depends on the oxidative pentose phosphate (PP) pathway to produce NADPH, its metabolic flux toward the oxidative PP pathway is limited due to the rigid glycolysis flux. To maximize NADPH supply for the isoprenoid production in yeast, upper glycolytic metabolic fluxes are reduced by introducing mutations into phosphofructokinase (PFK) along with overexpression of ZWF1 encoding glucose-6-phosphate (G6P) dehydrogenase. The PFK mutations (Pfk1 S724D and Pfk2 S718D) result in less glycerol production and more accumulation of G6P, which is a gateway metabolite toward the oxidative PP pathway. When combined with the PFK mutations, overexpression of ZWF1 caused substantial increases of [NADPH]/[NADP+] ratios whereas the effect of ZWF1 overexpression alone in the wild-type strain is not noticeable. Also, the introduction of ZWF1 overexpression and the PFK mutations into engineered yeast overexpressing acetyl-CoA C-acetyltransferase (ERG10), truncated HMG-CoA reductase isozyme 1 (tHMG1), and amorphadiene synthase (ADS) leads to a titer of 497 mg L–1 of amorphadiene (3.7-fold over the parental strain). These results suggest that perturbation of upper glycolytic fluxes, in addition to ZWF1 overexpression, is necessary for efficient NADPH supply through the oxidative PP pathway and enhanced production of isoprenoids by engineered S. cerevisiae.},
doi = {10.1002/biot.201900173},
journal = {Biotechnology Journal},
number = none,
volume = none,
place = {United States},
year = {2019},
month = {8}
}

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