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<p>Pyomelanin has been extensively applied in various fields. However, the yield of pyomelanin isolated from natural producers is low. Engineering microbial biosynthesis is considered a sustainable and economically feasible alternative. We utilize engineered <i>Komagataella phaffii</i> to increase the yield of homogentisic acid by 66-fold by balancing three different biosynthetic modules and further synthesizing pyomelanin by oxidative polymerization. During this process, we establish a pyomelanin color screening system and apply this system to perform directed evolution of 3-deoxy-D-arabinoheptulosonate 7-phosphate synthase and semirational design modification of hydroxyphenylpyruvate dioxygenase. We report the difference of pyomelanin between oxidative polymerization under alkaline conditions and that under laccase treatment. After high-density fermentation in a 5-L fermenter for 204 h, strain Pyo29 achieve the highest titer (70.5 ± 0.7 g/L). Finally, we demonstrate the potential application value of pyomelanin. This study provides a feasible solution for high-yield pyomelanin biosynthesis with substantial industrial production.</p>

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Biosynthesis of pyomelanin from methanol with engineered Komagataella phaffii and its characterizations

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年3月17日

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  • Xiangyu Zhu,
  • Jiawen Lin,
  • Shuli Liang,
  • Chenyi Qiu,
  • Yuchen Jiang,
  • Hengrui Liu,
  • Kun Zhu,
  • Qi Wang,
  • Chenfeng Du,
  • Yonghao Li,
  • Yuanhui Mao,
  • Huanbin Zheng,
  • Chengan Liu,
  • Yeyao Zhao,
  • Meirong An,
  • Yifeng Wu,
  • Jun Liu,
  • Xinying Zhang,
  • Ying Lin

摘要

Pyomelanin has been extensively applied in various fields. However, the yield of pyomelanin isolated from natural producers is low. Engineering microbial biosynthesis is considered a sustainable and economically feasible alternative. We utilize engineered Komagataella phaffii to increase the yield of homogentisic acid by 66-fold by balancing three different biosynthetic modules and further synthesizing pyomelanin by oxidative polymerization. During this process, we establish a pyomelanin color screening system and apply this system to perform directed evolution of 3-deoxy-D-arabinoheptulosonate 7-phosphate synthase and semirational design modification of hydroxyphenylpyruvate dioxygenase. We report the difference of pyomelanin between oxidative polymerization under alkaline conditions and that under laccase treatment. After high-density fermentation in a 5-L fermenter for 204 h, strain Pyo29 achieve the highest titer (70.5 ± 0.7 g/L). Finally, we demonstrate the potential application value of pyomelanin. This study provides a feasible solution for high-yield pyomelanin biosynthesis with substantial industrial production.