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Background <p>Protein purification is an essential step for complete characterization of any industrially important enzymes isolated from micro-organisms. In the present study, we have successfully isolated and identified fungal strain, <i>Penicillium sp</i>. (AL-3) for the bulk production of α-galactosidase enzyme which shows close phylogenetic resemblance with <i>Penicillium citrinum</i> (A79) using ITS sequencing (5.8&#xa0;S rRNA and 28&#xa0;S rRNA gene).</p> Methods and results <p>Selected fungus screened for its efficiency for α-galactosidase production using (1% w/v) soybean meal as a carbon substrate in culture media for maximum enzyme secretion. Enzyme was purified using ammonium sulphate precipitation (25–75%) with maximum enzyme activity of 2867.00 U/ml (yield of 90%). The eluted protein was further purified by DEAE ion exchange chromatography with an enzyme activity of 2260.04 U/mL (yield of 70.95%). The estimated molecular weight of the partially-purified enzyme was found to be 33&#xa0;kDa, using size exclusion chromatography and 81&#xa0;kDa using SDS-PAGE analysis which reflects its dimeric nature. Enzyme was inhibited by silver (Ag<sup>+</sup>) and subsequently by Hg<sup>+</sup> at 100 mM. However, Mg<sup>2+</sup> (454.29 at 20 mM) and EDTA (429.36 at 10 mM) and succinic acid (468.14 at 40 mM) act as enzyme inducers at 5 mM concentration, respectively. The enzyme had Km and Vmax values of 0.8 mM and 39&#xa0;mol/min/mg protein.</p> Conclusion <p><i>Penicillium citrinum</i> strain (A79) showed a high potential for the bulk production of the alpha-galactosidase with enzyme activity reported to be 733-745.15 U/ml under optimized culture conditions. Its properties make it widely applicable for digestive enzyme preparations, food, feed and medical industries.</p>

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Molecular characterization and protein purification of α-galactosidase from penicillium citrinum M(A79) for its industrial applications

  • 期刊论文

  • 发表日期：

    2025年7月23日

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  • Aishwarya Singh Chauhan,
  • Chitranjan Kumar,
  • Ritu Tiwari

摘要

Background

Protein purification is an essential step for complete characterization of any industrially important enzymes isolated from micro-organisms. In the present study, we have successfully isolated and identified fungal strain, Penicillium sp. (AL-3) for the bulk production of α-galactosidase enzyme which shows close phylogenetic resemblance with Penicillium citrinum (A79) using ITS sequencing (5.8 S rRNA and 28 S rRNA gene).

Methods and results

Selected fungus screened for its efficiency for α-galactosidase production using (1% w/v) soybean meal as a carbon substrate in culture media for maximum enzyme secretion. Enzyme was purified using ammonium sulphate precipitation (25–75%) with maximum enzyme activity of 2867.00 U/ml (yield of 90%). The eluted protein was further purified by DEAE ion exchange chromatography with an enzyme activity of 2260.04 U/mL (yield of 70.95%). The estimated molecular weight of the partially-purified enzyme was found to be 33 kDa, using size exclusion chromatography and 81 kDa using SDS-PAGE analysis which reflects its dimeric nature. Enzyme was inhibited by silver (Ag+) and subsequently by Hg+ at 100 mM. However, Mg2+ (454.29 at 20 mM) and EDTA (429.36 at 10 mM) and succinic acid (468.14 at 40 mM) act as enzyme inducers at 5 mM concentration, respectively. The enzyme had Km and Vmax values of 0.8 mM and 39 mol/min/mg protein.

Conclusion

Penicillium citrinum strain (A79) showed a high potential for the bulk production of the alpha-galactosidase with enzyme activity reported to be 733-745.15 U/ml under optimized culture conditions. Its properties make it widely applicable for digestive enzyme preparations, food, feed and medical industries.