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<p>Deep eutectic solvents (DESs) are promising green media for biocatalysis, yet their application in pyruvate decarboxylase (PDC)-mediated phenylacetylcarbinol (PAC) biotransformation with product separation remains limited. This study screened nine DESs using frozen-thawed whole cells (FT-WHC) of <i>Candida magnoliae</i> (<i>C. mag</i>.), followed by evaluation of water content and DES to phosphate buffer ratios. Glycerol-based DES resulted in significantly higher (<i>p</i> ≤ 0.05) volumetric PDC activity and PAC productivity than other DESs. Choline chloride: glycerol (ChCl: Gly (1:2) 20% (v/v) H<sub>2</sub>O) achieved the highest overall [PAC] of 148.2 ± 0.3 mM. Adding 100% (v/v) water decreased stability of volumetric PDC activity, but doubled PAC productivity, and reduced solvent cost compared with a 20% (v/v) H<sub>2</sub>O addition. The DES-to-phosphate buffer (Pi buffer) ratio at 1:1 was optimal, while lower ratios reduced PAC production. Spontaneous formation of PAC-enriched droplets (~ 2.5&#xa0;M) as a top phase indicated simplified downstream recovery. This indicates DES-based self-separation driven by polarity and product partitioning. Overall, ChCl: Gly (1:2) with 100% (v/v) H<sub>2</sub>O at a 1:1 DES: Pi buffer ratio was selected for future PAC biotransformation. These findings indicate that DESs enhance biocatalytic performance and enable integrated reaction and product separation in a single step, offering a sustainable route to biochemical production.</p>

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Spontaneous formation of highly concentrated phenylacetylcarbinol via in situ phase separation in deep eutectic solvents using Candida magnoliae whole cells

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年7月3日

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  • Su Lwin Htike,
  • Charin Techapun,
  • Yuthana Phimolsiripol,
  • Mohsen Gavahian,
  • Pornchai Rachtanapun,
  • Julaluk Khemacheewakul,
  • Suphat Phongthai,
  • Siraphat Taesuwan,
  • Sarinthip Thanakkasaranee,
  • Sumeth Sommanee,
  • Kritsadaporn Porninta,
  • Chatchadaporn Mahakuntha,
  • Juan Feng,
  • Rojarej Nunta,
  • Anbarasu Kumar,
  • Dan Gao,
  • Noppol Leksawasdi

摘要

Deep eutectic solvents (DESs) are promising green media for biocatalysis, yet their application in pyruvate decarboxylase (PDC)-mediated phenylacetylcarbinol (PAC) biotransformation with product separation remains limited. This study screened nine DESs using frozen-thawed whole cells (FT-WHC) of Candida magnoliae (C. mag.), followed by evaluation of water content and DES to phosphate buffer ratios. Glycerol-based DES resulted in significantly higher (p ≤ 0.05) volumetric PDC activity and PAC productivity than other DESs. Choline chloride: glycerol (ChCl: Gly (1:2) 20% (v/v) H2O) achieved the highest overall [PAC] of 148.2 ± 0.3 mM. Adding 100% (v/v) water decreased stability of volumetric PDC activity, but doubled PAC productivity, and reduced solvent cost compared with a 20% (v/v) H2O addition. The DES-to-phosphate buffer (Pi buffer) ratio at 1:1 was optimal, while lower ratios reduced PAC production. Spontaneous formation of PAC-enriched droplets (~ 2.5 M) as a top phase indicated simplified downstream recovery. This indicates DES-based self-separation driven by polarity and product partitioning. Overall, ChCl: Gly (1:2) with 100% (v/v) H2O at a 1:1 DES: Pi buffer ratio was selected for future PAC biotransformation. These findings indicate that DESs enhance biocatalytic performance and enable integrated reaction and product separation in a single step, offering a sustainable route to biochemical production.