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<p>With the increasing accumulation of plastic pollutants in various environments, research on microorganisms (including bacteria, fungi, and algae) with plastic degradation capabilities has gained significant attention. However, only a limited number of microbial plastic-degrading enzymes have been identified to date. This highlights that the degradation mechanisms employed by many plastic-degrading microorganisms, particularly filamentous fungi, remain insufficiently explored. In this study, we utilized a versatile fungal plasmid (pCT74) to express green fluorescent protein (GFP) in a marine-derived fungus <i>Alternaria alternata</i> strain FB1 with plastic degradation capabilities. Upon evaluating the degradation effect of polyester-type polyurethane (PU) film, we observed that different transformants exhibited three kinds of activities (the same, reduced, or enhanced degradation capability) compared to the FB1 wild-type strain. Further analysis of the plasmid fragment insertion sites in different transformants revealed that pCT74 integrates randomly into the genome of the host fungus. Notably, a direct correlation was found between the plasmid insertion site and the degradation capability of the corresponding transformant. Our findings not only redefine the potential applications of plasmid pCT74 in filamentous fungi but also show a novel research approach to identifying key enzymes involved in plastic degradation by fungi.</p>

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Mutant library construction and green fluorescent protein expression in a marine fungus capable of plastics biodegradation

  • 期刊论文

  • 发表日期：

    2025年8月18日

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  • Rui Liu,
  • Yani Wang,
  • Zhenjie Su,
  • Jun Yang,
  • Fan Fei,
  • Rongrong Gao,
  • Chaomin Sun

摘要

With the increasing accumulation of plastic pollutants in various environments, research on microorganisms (including bacteria, fungi, and algae) with plastic degradation capabilities has gained significant attention. However, only a limited number of microbial plastic-degrading enzymes have been identified to date. This highlights that the degradation mechanisms employed by many plastic-degrading microorganisms, particularly filamentous fungi, remain insufficiently explored. In this study, we utilized a versatile fungal plasmid (pCT74) to express green fluorescent protein (GFP) in a marine-derived fungus Alternaria alternata strain FB1 with plastic degradation capabilities. Upon evaluating the degradation effect of polyester-type polyurethane (PU) film, we observed that different transformants exhibited three kinds of activities (the same, reduced, or enhanced degradation capability) compared to the FB1 wild-type strain. Further analysis of the plasmid fragment insertion sites in different transformants revealed that pCT74 integrates randomly into the genome of the host fungus. Notably, a direct correlation was found between the plasmid insertion site and the degradation capability of the corresponding transformant. Our findings not only redefine the potential applications of plasmid pCT74 in filamentous fungi but also show a novel research approach to identifying key enzymes involved in plastic degradation by fungi.