Evidence map›Paper›PMID 41517837›Full record

ArticleBiotechnology and bioengineering2026

Engineering of 2-ketoacid Decarboxylases for Production of Isobutanol and Other Fusel Alcohols in Saccharomyces cerevisiae.

Joshua J Dietrich, Maelia Dziedzic, Jia Sun, Sri Harsha Adusumilli, Carla Gonçalves, Chris Todd Hittinger, Brian F Pfleger

Abstract read
In one paragraph

Article in Biotechnology and bioengineering, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

0numbers the graph read from it
0cells of the map it votes in
1citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

1 citing paper in PubMed.

  1. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

7 authors.

Joshua J DietrichDepartment of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, Wisconsin, USA.ORCID https://orcid.org/0000-0002-0047-6127
Maelia DziedzicDepartment of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, Wisconsin, USA.
Jia SunDepartment of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, Wisconsin, USA.
Sri Harsha AdusumilliDepartment of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, Wisconsin, USA.
Carla GonçalvesDepartment of Biological Sciences, Vanderbilt University, Nashville, Tennessee, USA.ORCID https://orcid.org/0000-0002-0420-4970
Chris Todd HittingerDOE Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, Wisconsin, USA.ORCID https://orcid.org/0000-0001-5088-7461
Brian F PflegerDepartment of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, Wisconsin, USA.

Funding

Biotechnology Training ProgramT32GM135066 · NIGMS · UNIVERSITY OF WISCONSIN-MADISON · PI SCOTT M. COYLE, BRIAN G FOX · 2020 to 2026
$7.0M
NIGMS NIH HHS T32 GM135066
6 · The paper itself

Abstract

Isobutanol is a fusel alcohol that can be produced microbially for use as a biofuel or upgraded into sustainable aviation fuel (SAF). A key enzyme in the isobutanol biosynthetic pathway is 2-ketoacid decarboxylase (KDC), which irreversibly decarboxylates 2-ketoisovalerate (KIV) to yield isobutyraldehyde. However, many previously characterized KDC enzymes also act promiscuously on other 2-ketoacids, (e.g., pyruvate) to produce a related aldehyde (e.g., acetaldehyde). This unwanted side reaction is especially important when isobutanol is produced in Saccharomyces cerevisiae (S. cerevisiae) because it leads to pyruvate being diverted to ethanol. In order to make S. cerevisiae a strict isobutanologen, a KDC enzyme that is specific for KIV must be deployed. In this study, we used a combination of cell-based and in vitro enzyme assays to investigate KDC substrate specificity, characterizing a large set of homologs for KIV, pyruvate, and phenylpyruvate (PPV) activity. A diverse range of substrate specificities was discovered, and some previously uncharacterized KDCs were revealed to have high KIV activity and low pyruvate activity. Multi-site saturation mutagenesis (SSM) of one of these KDCs identified mutants with increased KIV activity, while maintaining low levels of pyruvate activity. In a KIV bioconversion experiment, bioprospected and engineered KDCs allowed similar KIV consumption to when using the previously characterized Lactococcus lactis KdcA, though with some ethanol also produced. The KDCs identified here show promise for production of isobutanol and other alcohols derived from 2-ketoacids, and the dataset of newly characterized KDCs can inform future efforts to understand and engineer substrate specificity in KDCs.

Indexed as

ButanolsCarboxy-LyasesMetabolic EngineeringSaccharomyces cerevisiaeKeto AcidsSubstrate SpecificityButanolsCarboxy-Lyasesisobutyl alcoholKeto Acids2‐ketoacid decarboxylaseIsobutanolprotein engineeringSaccharomyces cerevisiaesite‐saturation mutagenesis

Identifiers

PMID41517837
PMCPMC13003428

What OpenQuestion holds

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.