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<p>Bixin, a natural carotenoid pigment derived from the seeds of <i>Bixa orellana</i> L. (annatto), is widely used as a food-grade colorant. Conventional extraction methods are often limited by poor mass transfer, incomplete pigment release, thermal or solvent-induced degradation, and concerns about residual solvent, collectively reducing extraction efficiency and hindering sustainable processing. This study evaluated enzyme-assisted extraction (EAE) of annatto seed pigments using amylase, cellulase, and pectinase, each tested individually, across controlled extraction temperatures ranging from 40 to 70&#xa0;°C and enzyme concentrations from 1 to 10% (w/v). A Central Composite Design comprising 13 runs per enzyme with triplicate experiments (<i>n</i> = 3; total 117 extractions) assessed the effects of temperature and enzyme loading on bixin and norbixin yields, colour attributes, phenolic content, and antioxidant activity. Significant effects (<i>p</i> &lt; 0.01) of both temperature and enzyme concentration were observed for most responses, and all fitted RSM models demonstrated high adequacy (R² = 0.96–0.99; CV &lt; 1%) with non-significant lack-of-fit. The optimized condition of 60&#xa0;°C with 10% cellulase concentration yielded maximum bixin (3.92 ± 1.61) and norbixin (0.91 ± 0.64), with colour attributes a* (25.32 ± 0.02) and Chroma (35.08 ± 1.13), a TPC of 43.33 ± 0.24&#xa0;mg GAE/g, and antioxidant activity of 44.63 ± 0.07%. DSC analysis indicated pigment transitions with an onset at 80.9&#xa0;°C and a peak at 108.6&#xa0;°C. Incorporation of the extract into butter demonstrated strong chromatic stability. These results confirm that EAE is an effective approach for enhancing annatto pigment recovery and improving its functional properties.</p> Graphical Abstract <p></p>

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Sustainable enzyme-assisted extraction of bixin and norbixin from Bixa orellana seeds: Evaluation of yield, quality attributes, and microstructural changes

  • 期刊论文

  • 发表日期：

    2026年1月18日

查看全文
  • Nila Shireen Baburaj,
  • Sudha Paramadhas,
  • S. Ganga Kishore,
  • R. Rahul,
  • Meyyammai Annamalai,
  • Balakrishnan Murugesan,
  • Preetha Palanisamy,
  • Madhuresh Dwivedi

摘要

Bixin, a natural carotenoid pigment derived from the seeds of Bixa orellana L. (annatto), is widely used as a food-grade colorant. Conventional extraction methods are often limited by poor mass transfer, incomplete pigment release, thermal or solvent-induced degradation, and concerns about residual solvent, collectively reducing extraction efficiency and hindering sustainable processing. This study evaluated enzyme-assisted extraction (EAE) of annatto seed pigments using amylase, cellulase, and pectinase, each tested individually, across controlled extraction temperatures ranging from 40 to 70 °C and enzyme concentrations from 1 to 10% (w/v). A Central Composite Design comprising 13 runs per enzyme with triplicate experiments (n = 3; total 117 extractions) assessed the effects of temperature and enzyme loading on bixin and norbixin yields, colour attributes, phenolic content, and antioxidant activity. Significant effects (p < 0.01) of both temperature and enzyme concentration were observed for most responses, and all fitted RSM models demonstrated high adequacy (R² = 0.96–0.99; CV < 1%) with non-significant lack-of-fit. The optimized condition of 60 °C with 10% cellulase concentration yielded maximum bixin (3.92 ± 1.61) and norbixin (0.91 ± 0.64), with colour attributes a* (25.32 ± 0.02) and Chroma (35.08 ± 1.13), a TPC of 43.33 ± 0.24 mg GAE/g, and antioxidant activity of 44.63 ± 0.07%. DSC analysis indicated pigment transitions with an onset at 80.9 °C and a peak at 108.6 °C. Incorporation of the extract into butter demonstrated strong chromatic stability. These results confirm that EAE is an effective approach for enhancing annatto pigment recovery and improving its functional properties.

Graphical Abstract