Microorganism for fermentative production of 2-phenylethanol from gaseous substrates
Inventors
Liew, Fungmin • Koepke, Michael • Nagaraju, Shilpa • Harris, Audrey
Assignees
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Publication Number
US-12234492-B2
Publication Date
2025-02-25
Expiration Date
Abstract
Disclosed herein are improved methods for production of 2-phenylethanol by microbial fermentation of substrates comprising carbon monoxide and/or carbon dioxide and further disclosed are genetically modified microorganisms for use in such methods that alleviate dependence on natural and petrochemical processes.
Core Innovation
The invention relates to C1-fixing Wood-Ljungdahl microorganisms engineered to produce 2-phenylethanol from gaseous C1 feeds containing CO and/or CO2 and optionally H2. The Wood-Ljungdahl pathway converts the gaseous C1 inputs into central carbon metabolism, and 2-phenylethanol formation is routed through phenylpyruvate and phenylacetaldehyde intermediates. The document focuses on production using C1 carbon sources such as syngas and industrial waste gas.
Core pathway engineering includes a heterologous phenylpyruvate decarboxylase that converts phenylpyruvate to phenylacetaldehyde and a heterologous phenylacetaldehyde reductase that converts phenylacetaldehyde to 2-phenylethanol. The claimed microorganism comprises these heterologous enzymes, each encoded by nucleic acids having at least 90% identity to specified SEQ ID NO sets. The disclosure further enumerates heterologous enzyme and gene options and combinations, including decarboxylase variants and multiple phenylacetaldehyde reductase variants.
The disclosure also describes optional additional heterologous shikimate-pathway and chorismate/phenylpyruvate enzymes to improve metabolic flux. Reported constraints emphasize reduced or absent formation of other C3+ alcohol byproducts, with a selectivity framing around desired 2-phenylethanol. The document includes experimental content addressing 2-phenylethanol tolerance issues, comparative selectivity effects between decarboxylase variants, PAR variant screening, combinatorial assembly and screening of 2-PE pathway libraries, and continuous process performance improvements for selected combinatorial strains using pretreated real syngas.
Claims Coverage
The patent includes one independent claim. The independent claim is directed to a Wood-Ljungdahl C1-fixing microorganism that produces 2-phenylethanol via two heterologous enzymatic conversions, with each heterologous enzyme defined by at least 90% identity to specified SEQ ID sets. Dependent claims refine the context, constrain product and byproduct profiles, and specify example heterologous enzyme and gene sets and combinatorial-strain encoding constraints.
Wood-Ljungdahl 2-phenylethanol production via decarboxylase and phenylacetaldehyde conversion
A C1-fixing Wood-Ljungdahl microorganism that produces 2-phenylethanol, wherein the microorganism comprises a heterologous decarboxylase converting phenylpyruvate to phenylacetaldehyde encoded by a nucleic acid with at least 90% identity to SEQ ID NO: 1, 2, or 4, and a heterologous phenylacetaldehyde enzyme converting phenylacetaldehyde to 2-phenylethanol encoded by a nucleic acid with at least 90% identity to SEQ ID NO: 3, 5, 6, 7, 8, or 9.
Overall claim coverage is focused on a Wood-Ljungdahl C1-fixing microorganism producing 2-phenylethanol through heterologous phenylpyruvate decarboxylase and heterologous phenylacetaldehyde conversion to 2-phenylethanol, with each heterologous enzyme sequence identity constrained to specified SEQ ID sets. Dependent refinements cover particular gaseous C1 feed options, additional selectivity constraints on C3-C10 alcohols, and specification of example heterologous enzyme and gene sets and combinatorial-strain encoding.
Stated Advantages
Enables production of 2-phenylethanol from gaseous C1 feeds using Wood-Ljungdahl microorganisms.
Supports selectivity by constraining production to 2-phenylethanol and excluding other C3-C10 alcohols beyond those specified.
Provides improved performance in continuous operation with selected combinatorial strains using pretreated real syngas.
Documented Applications
Producing 2-phenylethanol by culturing the claimed C1-fixing Wood-Ljungdahl microorganism with a gaseous C1-carbon source containing CO, CO2, and/or H2.
Use of gaseous substrates including syngas or industrial waste gas for producing 2-phenylethanol.
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