14 Engineering Saccharomyces cerevisiae for Production of Fatty
Acids and Their Derivatives
LEONIE BAUMANN
1
, FLORIAN WERNIG
1 , SANDRA BORN
1
, MISLAV OREB
1
CONTENTS
I. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 339
II. Providing Carbon, Redox Power, and Energy
for Fatty Acid Synthesis: The Options for
Precursor Supply Routes . . . . . . . . . . . . . . . . . . . 340
A. Engineering the Pyruvate Dehydrogenase
Bypass . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 341
B. ATP-Independent Pyruvate-to-Acetyl-CoA
Routes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 343
C. Phosphoketolase Pathway . . . . . . . . . . . . . . . . 344
D. Citrate-Oxaloacetate Shuttle . . . . . . . . . . . . . . 345
E. Strategies to Manipulate NADPH Level
Independently of Precursor Supply Routes346
F. Engineering Malonyl-CoA Supply . . . . . . . . 346
III. Strategies for Chain Length Control . . . . . . . . 348
A. Manipulation of the Natural Fatty Acid
Biosynthesis and Elongation Machinery . . 348
B. Reversal of b-Oxidation as an Orthogonal
Pathway for Fatty Acid Biosynthesis . . . . . . 349
IV. Chassis Engineering . . . . . . . . . . . . . . . . . . . . . . . . 351
A. Elimination of By-product Formation and
Fatty Acid Degradation . . . . . . . . . . . . . . . . . . . 351
B. Engineering Yeast Product Tolerance and
Excretion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 352
C. Dynamic Pathway Control and Biosensors354
V. S. cerevisiae as a Production Platform for
Fatty Acid Derivatives . . . . . . . . . . . . . . . . . . . . . . 356
A. Alkenes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 357
B. Dicarboxylic Acids . . . . . . . . . . . . . . . . . . . . . . . 357
C. Fatty Aldehydes, Alkanes, and Alcohols . . 357
D. Wax Esters and Fatty Acid Ethyl Esters . . 358
VI. Optimization of Fermentation
Conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 359
VII. Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 360
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 361
Abbreviations
AcCoA
Acetyl-CoA
AcP
Acetyl-phosphate
ALE
Adaptive laboratory evolution
CoA
Coenzyme A
FA
Fatty acid(s)
FACS
Fluorescence-activated cell sorting
FAEE
Fatty acid ethyl ester(s)
GAP
Glyceraldehyde-3-phosphate
gTME
Global transcription machinery engineering
MalCoA Malonyl-CoA
PPP
Pentose phosphate pathway
SE
Steryl ester
TAG
Triacylglycerol
TCA
Tricarboxylic acid
TE
Thioesterase
X5P
Xylulose 5-phosphate
I. Introduction
Fatty acids (FAs) and their derivatives, such as
fatty alcohols, dicarboxylic acids, FA esters,
alkanes, and alkenes, are frequently summarized under the generic term “oleochemicals.”
The diversity of physicochemical properties of
oleochemicals is determined by their functional
groups and by the length of the aliphatic chains.
This is reflected by a variety of products containing these classes of compounds, including
fuels, lubricants, surfactants, detergents, cosmetics, food additives, and pharmaceuticals.
Consequently, the production volume of different oleochemicals is measured in millions of
1 Faculty of Biological Sciences, Institute of Molecular
Biosciences, Goethe-University Frankfurt, Frankfurt,
Germany; e-mail: M.Oreb@bio.uni-frankfurt.de
Genetics and Biotechnology, 3 rd Edition
The Mycota II
J.P. Benz, K. Schipper (Eds.)
© Springer Nature Switzerland AG 2020