Scholarlink

<b>Abstract</b>— <p>Cholesterol oxidase (ChO) belongs to the family of flavin-containing enzymes that catalyze the oxidation of 3β-hydroxy-5-ene steroids with the formation of hydrogen peroxide and the products with a 3‑keto-4-ene structure. This study dealt with obtaining a secreted mature form of heterologous ChO from the actinobacterium <i>Nocardioides simplex</i> (ChO<sub><i>Ns</i></sub>) in the cells of a recombinant methylotrophic yeast <i>Pichia pastoris</i> GS115. Based on the plasmid vector pPICZαA, genetic constructs were designed for heterologous expression of the target gene <i>choN</i> (<i>KR76_09550</i>) from <i>N. simplex</i> in recombinant yeast. We compared the activity of the cell-free culture supernatant of recombinant <i>P. pastoris</i> strains containing secreted heterologous ChO<sub><i>Ns</i></sub> with respect to the conversion of cholesterol to cholestenone, depending on the presence and position of a 6 × His tag in the ChO<sub><i>Ns</i></sub> molecule. The conditions for synthesis of the most active heterologous secreted ChO<sub><i>Ns</i></sub> in the selected recombinant yeast strain were optimized.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Synthesis of Secreted Heterologous Actinobacterial Cholesterol Oxidase from Nocardioides simplex in Cells of Methylotrophic Yeast Pichia pastoris (Komagataella phaffii) GS115

  • 期刊论文

  • 发表日期：

    2025年12月1日

查看全文
  • V. V. Kollerov,
  • V. V. Fokina,
  • M. V. Karpov,
  • A. A. Shutov,
  • M. V. Donova

摘要

Abstract—

Cholesterol oxidase (ChO) belongs to the family of flavin-containing enzymes that catalyze the oxidation of 3β-hydroxy-5-ene steroids with the formation of hydrogen peroxide and the products with a 3‑keto-4-ene structure. This study dealt with obtaining a secreted mature form of heterologous ChO from the actinobacterium Nocardioides simplex (ChONs) in the cells of a recombinant methylotrophic yeast Pichia pastoris GS115. Based on the plasmid vector pPICZαA, genetic constructs were designed for heterologous expression of the target gene choN (KR76_09550) from N. simplex in recombinant yeast. We compared the activity of the cell-free culture supernatant of recombinant P. pastoris strains containing secreted heterologous ChONs with respect to the conversion of cholesterol to cholestenone, depending on the presence and position of a 6 × His tag in the ChONs molecule. The conditions for synthesis of the most active heterologous secreted ChONs in the selected recombinant yeast strain were optimized.