as metabolic imbalances and product toxicity
have been tackled, yields, titers, and productivity could reach economically viable levels
before long.
Acknowledgments L.B., F.W., and M.O. received funding from the European Union’s Horizon 2020 research
and innovation program under Grant Agreement No.
720824. S.B. received funding from the German Federal
Ministry of Food and Agriculture following a decision
of the German Bundestag under the Grant Number
22026315. The responsibility for the content of this
publication lies with the authors. The authors thank
Eckhard Boles for helpful comments on the manuscript.
References
Akhtar MK, Turner NJ, Jones PR (2013) Carboxylic acid
reductase is a versatile enzyme for the conversion
of fatty acids into fuels and chemical commodities.
Proc Natl Acad Sci U S A 110:87–92
Alexandre H, Mathieu B, Charpentier C (1996) Alteration in membrane fluidity and lipid composition,
and modulation of H+-ATPase activity in Saccharomyces cerevisiae caused by decanoic acid. Microbiology 142:469–475
Alper H, Fischer C, Nevoigt E, Stephanopoulos G (2005)
Tuning genetic control through promoter engineering. Proc Natl Acad Sci U S A 102:12678–
12683
Alper H, Moxley J, Nevoigt E, Fink GR, Stephanopoulos
G (2006) Engineering yeast transcription machinery for improved ethanol tolerance and production. Science 314:1565–1568
Baganz F, Hayes A, Marren D, Gardner DCJ, Oliver SG
(1997) Suitability of replacement markers for
functional analysis studies in Saccharomyces cerevisiae. Yeast 13:1563–1573
Basso TO, Dario MG, Tonso A, Stambuk BU, Gombert
AK (2010) Insufficient uracil supply in fully aerobic chemostat cultures of Saccharomyces cerevisiae
leads to respiro-fermentative metabolism and
double nutrient-limitation. Biotechnol Lett
32:973–977
Baumann L, Rajkumar AS, Morrissey JP, Boles E, Oreb
M (2018) A yeast-based biosensor for screening of
short- and medium-chain fatty acid production.
ACS Synth Biol 7:2640–2646
Becker S, Schmoldt H-U, Adams TM, Wilhelm S, Kolmar H (2004) Ultra-high-throughput screening
based on cell-surface display and fluorescenceactivated cell sorting for the identification of
novel biocatalysts. Curr Opin Biotechnol 15:323–
329
Beopoulos A, Chardot T, Nicaud J-M (2009) Yarrowia
lipolytica: a model and a tool to understand the
mechanisms implicated in lipid accumulation.
Biochimie 91:692–696
Besada-Lombana PB, Fernandez-Moya R, Fenster J, Da
Silva NA (2017) Engineering Saccharomyces cerevisiae fatty acid composition for increased tolerance to octanoic acid. Biotechnol Bioeng
114:1531–1538
Black PN, DiRusso CC (2007) Yeast acyl-CoA synthetases at the crossroads of fatty acid metabolism
and regulation. Biochim Biophys Acta 1771:286–
298
Bocanegra JA, Scrutton NS, Perham RN (1993) Creation
of an NADP-dependent pyruvate dehydrogenase
multienzyme complex by protein engineering.
Biochemistry 32:2737–2740
Brignole EJ, Smith S, Asturias FJ (2009) Conformational
flexibility of metazoan fatty acid synthase enables
catalysis. Nat Struct Mol Biol 16:190–197
Buijs NA, Zhou YJ, Siewers V, Nielsen J (2015) Longchain alkane production by the yeast Saccharomyces cerevisiae. Biotechnol Bioeng 112:1275–1279
C ¸ akar ZP, Sauer U, Bailey JE (1999) Metabolic engineering of yeast: the perils of auxotrophic hosts. Biotechnol Lett 21:611–616
Cardenas J, Da Silva NA (2016) Engineering cofactor
and transport mechanisms in Saccharomyces cerevisiae for enhanced acetyl-CoA and polyketide
biosynthesis. Metab Eng 36:80–89
Caspeta L, Chen Y, Ghiaci P, Feizi A, Buskov S, Hallstro ¨m BM, Petranovic D, Nielsen J (2014) Altered
sterol composition renders yeast thermotolerant.
Science 346:75–78
Cernak P, Estrela R, Poddar S, Skerker JM, Cheng Y-F,
Carlson AK, Chen B, Glynn VM, Furlan M, Ryan
OW, Donnelly MK, Arkin AP, Taylor JW, Cate JHD
(2018) Engineering Kluyveromyces marxianus as a
robust synthetic biology platform host. MBio 9:1–
16
Chen B, Ling H, Chang MW (2013a) Transporter engineering for improved tolerance against alkane biofuels in Saccharomyces cerevisiae. Biotechnol
Biofuels 6:21
Chen Y, Daviet L, Schalk M, Siewers V, Nielsen J
(2013b) Establishing a platform cell factory
through engineering of yeast acetyl-CoA metabolism. Metab Eng 15:48–54
Chen B, Lee D-Y, Chang MW (2015) Combinatorial
metabolic engineering of Saccharomyces cerevisiae
for terminal alkene production. Metab Eng 31:53–
61
Choi JW, Da Silva NA (2014) Improving polyketide and
fatty acid synthesis by engineering of the yeast
acetyl-CoA carboxylase. J Biotechnol 187:56–59
Clomburg JM, Vick JE, Blankschien MD, Rodrı ´guezMoya ´ M, Gonzalez R (2012) A synthetic biology
approach to engineer a functional reversal of the
b-oxidation cycle. ACS Synth Biol 1:541–554
Clomburg JM, Blankschien MD, Vick JE, Chou A, Kim
S, Gonzalez R (2015) Integrated engineering of b14 Engineering Saccharomyces cerevisiae for Production of Fatty Acids and Their Derivatives
361
have been tackled, yields, titers, and productivity could reach economically viable levels
before long.
Acknowledgments L.B., F.W., and M.O. received funding from the European Union’s Horizon 2020 research
and innovation program under Grant Agreement No.
720824. S.B. received funding from the German Federal
Ministry of Food and Agriculture following a decision
of the German Bundestag under the Grant Number
22026315. The responsibility for the content of this
publication lies with the authors. The authors thank
Eckhard Boles for helpful comments on the manuscript.
References
Akhtar MK, Turner NJ, Jones PR (2013) Carboxylic acid
reductase is a versatile enzyme for the conversion
of fatty acids into fuels and chemical commodities.
Proc Natl Acad Sci U S A 110:87–92
Alexandre H, Mathieu B, Charpentier C (1996) Alteration in membrane fluidity and lipid composition,
and modulation of H+-ATPase activity in Saccharomyces cerevisiae caused by decanoic acid. Microbiology 142:469–475
Alper H, Fischer C, Nevoigt E, Stephanopoulos G (2005)
Tuning genetic control through promoter engineering. Proc Natl Acad Sci U S A 102:12678–
12683
Alper H, Moxley J, Nevoigt E, Fink GR, Stephanopoulos
G (2006) Engineering yeast transcription machinery for improved ethanol tolerance and production. Science 314:1565–1568
Baganz F, Hayes A, Marren D, Gardner DCJ, Oliver SG
(1997) Suitability of replacement markers for
functional analysis studies in Saccharomyces cerevisiae. Yeast 13:1563–1573
Basso TO, Dario MG, Tonso A, Stambuk BU, Gombert
AK (2010) Insufficient uracil supply in fully aerobic chemostat cultures of Saccharomyces cerevisiae
leads to respiro-fermentative metabolism and
double nutrient-limitation. Biotechnol Lett
32:973–977
Baumann L, Rajkumar AS, Morrissey JP, Boles E, Oreb
M (2018) A yeast-based biosensor for screening of
short- and medium-chain fatty acid production.
ACS Synth Biol 7:2640–2646
Becker S, Schmoldt H-U, Adams TM, Wilhelm S, Kolmar H (2004) Ultra-high-throughput screening
based on cell-surface display and fluorescenceactivated cell sorting for the identification of
novel biocatalysts. Curr Opin Biotechnol 15:323–
329
Beopoulos A, Chardot T, Nicaud J-M (2009) Yarrowia
lipolytica: a model and a tool to understand the
mechanisms implicated in lipid accumulation.
Biochimie 91:692–696
Besada-Lombana PB, Fernandez-Moya R, Fenster J, Da
Silva NA (2017) Engineering Saccharomyces cerevisiae fatty acid composition for increased tolerance to octanoic acid. Biotechnol Bioeng
114:1531–1538
Black PN, DiRusso CC (2007) Yeast acyl-CoA synthetases at the crossroads of fatty acid metabolism
and regulation. Biochim Biophys Acta 1771:286–
298
Bocanegra JA, Scrutton NS, Perham RN (1993) Creation
of an NADP-dependent pyruvate dehydrogenase
multienzyme complex by protein engineering.
Biochemistry 32:2737–2740
Brignole EJ, Smith S, Asturias FJ (2009) Conformational
flexibility of metazoan fatty acid synthase enables
catalysis. Nat Struct Mol Biol 16:190–197
Buijs NA, Zhou YJ, Siewers V, Nielsen J (2015) Longchain alkane production by the yeast Saccharomyces cerevisiae. Biotechnol Bioeng 112:1275–1279
C ¸ akar ZP, Sauer U, Bailey JE (1999) Metabolic engineering of yeast: the perils of auxotrophic hosts. Biotechnol Lett 21:611–616
Cardenas J, Da Silva NA (2016) Engineering cofactor
and transport mechanisms in Saccharomyces cerevisiae for enhanced acetyl-CoA and polyketide
biosynthesis. Metab Eng 36:80–89
Caspeta L, Chen Y, Ghiaci P, Feizi A, Buskov S, Hallstro ¨m BM, Petranovic D, Nielsen J (2014) Altered
sterol composition renders yeast thermotolerant.
Science 346:75–78
Cernak P, Estrela R, Poddar S, Skerker JM, Cheng Y-F,
Carlson AK, Chen B, Glynn VM, Furlan M, Ryan
OW, Donnelly MK, Arkin AP, Taylor JW, Cate JHD
(2018) Engineering Kluyveromyces marxianus as a
robust synthetic biology platform host. MBio 9:1–
16
Chen B, Ling H, Chang MW (2013a) Transporter engineering for improved tolerance against alkane biofuels in Saccharomyces cerevisiae. Biotechnol
Biofuels 6:21
Chen Y, Daviet L, Schalk M, Siewers V, Nielsen J
(2013b) Establishing a platform cell factory
through engineering of yeast acetyl-CoA metabolism. Metab Eng 15:48–54
Chen B, Lee D-Y, Chang MW (2015) Combinatorial
metabolic engineering of Saccharomyces cerevisiae
for terminal alkene production. Metab Eng 31:53–
61
Choi JW, Da Silva NA (2014) Improving polyketide and
fatty acid synthesis by engineering of the yeast
acetyl-CoA carboxylase. J Biotechnol 187:56–59
Clomburg JM, Vick JE, Blankschien MD, Rodrı ´guezMoya ´ M, Gonzalez R (2012) A synthetic biology
approach to engineer a functional reversal of the
b-oxidation cycle. ACS Synth Biol 1:541–554
Clomburg JM, Blankschien MD, Vick JE, Chou A, Kim
S, Gonzalez R (2015) Integrated engineering of b14 Engineering Saccharomyces cerevisiae for Production of Fatty Acids and Their Derivatives
361