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<p>Natural deep eutectic solvents (NADES) have previously shown considerable impact in improving enzyme activity. NADES can compensate for some of the main disadvantages of traditional organic solvents and ionic liquids and meet all requirements of green chemistry. Microbial transglutaminase (MTG) is widely used in the food industry, and its industrial usage is rising rapidly. This study investigated MTG’s activity, thermal stability, and tertiary structure in betaine-glycerol and choline chloride-glycerol NADES. The results indicate that choline chloride-based eutectics exhibited a better media for MTG activity and stability than the aqueous buffer and betaine-based NADES. MTG exhibited its highest activity in a 10% (v/v) glycerol:choline chloride (2:1) mixture (Gly2C). The enzyme also showed enhanced thermal stability, with a half-life of 173&#xa0;min at 50&#xa0;°C in Gly2C, compared to 18&#xa0;&#xa0;min in Tris buffer and 69&#xa0;&#xa0;min in Gly2B. At 60&#xa0;°C, the enzyme’s half-life was extended in NADES, with the longest stability observed in Gly2C. Associated conformational changes caused by solvents were monitored using the fluorescence technique. This is the first study describing MTG’s activity, stability, and structural changes in NADES.</p>

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Enhancing transglutaminase activity and thermal stability using natural deep eutectic solvents

  • 期刊论文

  • 发表日期：

    2025年11月3日

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  • Azita Tafreshi-Hosseini,
  • Masoud Giahi,
  • Bahareh Dabirmanesh,
  • Yasaman Azizpour,
  • Khosro Khajeh

摘要

Natural deep eutectic solvents (NADES) have previously shown considerable impact in improving enzyme activity. NADES can compensate for some of the main disadvantages of traditional organic solvents and ionic liquids and meet all requirements of green chemistry. Microbial transglutaminase (MTG) is widely used in the food industry, and its industrial usage is rising rapidly. This study investigated MTG’s activity, thermal stability, and tertiary structure in betaine-glycerol and choline chloride-glycerol NADES. The results indicate that choline chloride-based eutectics exhibited a better media for MTG activity and stability than the aqueous buffer and betaine-based NADES. MTG exhibited its highest activity in a 10% (v/v) glycerol:choline chloride (2:1) mixture (Gly2C). The enzyme also showed enhanced thermal stability, with a half-life of 173 min at 50 °C in Gly2C, compared to 18  min in Tris buffer and 69  min in Gly2B. At 60 °C, the enzyme’s half-life was extended in NADES, with the longest stability observed in Gly2C. Associated conformational changes caused by solvents were monitored using the fluorescence technique. This is the first study describing MTG’s activity, stability, and structural changes in NADES.