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<p>Polyhydroxyalkanoates are a microbial storage polymer that presents similar characteristics to plastics, therefore are usually referred to as <i>bioplastics</i>. The key enzyme involved in the synthesis of polyhydroxyalkanoates is called PHA synthase, but it is always forming a synthesis cassette of three to five enzymes. In this work, we present a novel class III polyhydroxyalkanoate synthesis cassette identified using a sequence-based metagenomics approach. From two metagenomes from dairy industries stabilization ponds, we found four complete synthesis cassettes. One of them was successfully cloned into <i>Escherichia coli</i> CGSC: 6576, a bacterial strain able to grow in lactose and whey permeate, a by-product of dairy industries that represent an environmental risk. This recombinant strain grown in lactose was able to accumulate nearly 40% of dry weight as insoluble material identified by Fourier Transform Infrared Spectroscopy (FT-IR), Gas Chromatography with Flame Ionization Detection (GC-FID) and characterized by Nuclear Magnetic Resonance (NMR) as poly(3-hydroxybutyrate), one of the main polyhydroxyalkanoates commercially used. This research presents the complete process, from the identification of novel enzymes from uncultured microorganisms to the synthesis at laboratory scale of compounds of industrial interest.</p>

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Evaluation of a Class III PHA Synthase Identified in Dairy Pond Metagenomes for Biopolymer Production

  • 期刊论文

  • 发表日期：

    2026年1月12日

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  • J. M. Irazoqui,
  • M. E. Mainez,
  • A. Valenti,
  • G. M. Santiago,
  • R. Paez,
  • A. F. Amadio,
  • M. F. Eberhardt

摘要

Polyhydroxyalkanoates are a microbial storage polymer that presents similar characteristics to plastics, therefore are usually referred to as bioplastics. The key enzyme involved in the synthesis of polyhydroxyalkanoates is called PHA synthase, but it is always forming a synthesis cassette of three to five enzymes. In this work, we present a novel class III polyhydroxyalkanoate synthesis cassette identified using a sequence-based metagenomics approach. From two metagenomes from dairy industries stabilization ponds, we found four complete synthesis cassettes. One of them was successfully cloned into Escherichia coli CGSC: 6576, a bacterial strain able to grow in lactose and whey permeate, a by-product of dairy industries that represent an environmental risk. This recombinant strain grown in lactose was able to accumulate nearly 40% of dry weight as insoluble material identified by Fourier Transform Infrared Spectroscopy (FT-IR), Gas Chromatography with Flame Ionization Detection (GC-FID) and characterized by Nuclear Magnetic Resonance (NMR) as poly(3-hydroxybutyrate), one of the main polyhydroxyalkanoates commercially used. This research presents the complete process, from the identification of novel enzymes from uncultured microorganisms to the synthesis at laboratory scale of compounds of industrial interest.