<p>Reconstructing the precise biosynthesis of structurally complex natural esters, such as monoterpene esters, in engineered microbes remains a major challenge, owing to the limited repertoire of highly selective alcohol acyltransferases and the lack of compatible pathway modularity. Here, we establish a dual-substrate microbial platform to profile the activities of alcohol acyltransferase (AAT) to synthesize three distinct classes of monoterpene esters: monoterpenyl esters, monoterpenoate esters, and monoterpenyl monoterpenoate esters, enabling access to both natural and non-natural monoterpene ester biosynthetic pathways. Through structure-guided critical residue engineering and dual-substrate molar ratio tuning, we achieve selective biosynthesis of &gt;C2 acyl-CoA-derived monoterpene esters, despite competing intracellular acetyl-CoA. Coculture engineering further redistributed metabolic fluxes between acyl-CoA and alcohol precursors, yielding 11.50 g/L linalyl acetate and 3.16 g/L geranyl butyrate in 1-L bioreactor. This study expands the biosynthetic space of monoterpene esters and provides a versatile strategy to control AAT selectivity, offering a plug-and-play, scalable framework for ester biomanufacturing.</p>

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A microbial platform for selective biosynthesis of monoterpene esters

  • Dianqi Yang,
  • Hong Liang,
  • Xuxu Li,
  • Bowen Wang,
  • Chenyu Zhang,
  • Mengyao Yuan,
  • Kang Zhou,
  • Zhiyong Cui,
  • Panhao Niu,
  • Yi-Lei Zhao,
  • Yuan Liu,
  • Zuobing Xiao,
  • Lianzhong Ai,
  • Yongjin J. Zhou,
  • Baoguo Sun,
  • Xiaoqiang Ma

摘要

Reconstructing the precise biosynthesis of structurally complex natural esters, such as monoterpene esters, in engineered microbes remains a major challenge, owing to the limited repertoire of highly selective alcohol acyltransferases and the lack of compatible pathway modularity. Here, we establish a dual-substrate microbial platform to profile the activities of alcohol acyltransferase (AAT) to synthesize three distinct classes of monoterpene esters: monoterpenyl esters, monoterpenoate esters, and monoterpenyl monoterpenoate esters, enabling access to both natural and non-natural monoterpene ester biosynthetic pathways. Through structure-guided critical residue engineering and dual-substrate molar ratio tuning, we achieve selective biosynthesis of >C2 acyl-CoA-derived monoterpene esters, despite competing intracellular acetyl-CoA. Coculture engineering further redistributed metabolic fluxes between acyl-CoA and alcohol precursors, yielding 11.50 g/L linalyl acetate and 3.16 g/L geranyl butyrate in 1-L bioreactor. This study expands the biosynthetic space of monoterpene esters and provides a versatile strategy to control AAT selectivity, offering a plug-and-play, scalable framework for ester biomanufacturing.