Abstract
Microorganisms represent the most abundant biomass on the planet; however, because of several cultivation technique limitations, most of this genetic patrimony has been inaccessible. Due to the advent of metagenomic methodologies, such limitations have been overcome. Prevailing over these limitations enabled the genetic pool of non-cultivable microorganisms to be exploited for improvements in the development of biotechnological products. By utilising a metagenomic approach, we identified a new gene related to biosurfactant production and hydrocarbon degradation. Environmental DNA was extracted from soil samples collected on the banks of the Jundiaí River (Natal, Brazil), and a metagenomic library was constructed. Functional screening identified the clone 3C6, which was positive for the biosurfactant protein and revealed an open reading frame (ORF) with high similarity to sequences encoding a hypothetical protein from species of the family Halobacteriaceae. This protein was purified and exhibited biosurfactant activity. Due to these properties, this protein was named metagenomic biosurfactant protein 1 (MBSP1). In addition, E. coli RosettaTM (DE3) strain cells transformed with the MBSP1 clone showed an increase in aliphatic hydrocarbon degradation. In this study, we described a single gene encoding a protein with marked tensoactive properties that can be produced in a host cell, such as Escherichia coli, without substrate dependence. Furthermore, MBSP1 has been demonstrated as the first protein with these characteristics described in the Archaea or Bacteria domains.
Highlights
Microorganisms represent the most abundant biomass on the planet; because of several cultivation technique limitations, most of this genetic patrimony has been inaccessible
We described the functional characterisation of the first open reading frame (ORF) (Supplementary Fig. S1)
The 897 bp ORF encodes a polypeptide of 298 amino acids with an estimated weight of 31 kDa and a theoretical isoelectric point of 4.40. This sequence showed a high similarity with hypothetical proteins of the family Halobacteriaceae
Summary
Microorganisms represent the most abundant biomass on the planet; because of several cultivation technique limitations, most of this genetic patrimony has been inaccessible. Due to the advent of metagenomic methodologies, such limitations have been overcome Prevailing over these limitations enabled the genetic pool of non-cultivable microorganisms to be exploited for improvements in the development of biotechnological products. Functional screening identified the clone 3C6, which was positive for the biosurfactant protein and revealed an open reading frame (ORF) with high similarity to sequences encoding a hypothetical protein from species of the family Halobacteriaceae. This protein was purified and exhibited biosurfactant activity. Biosurfactant properties have been shown to exhibit emulsification, de-emulsification, dispersion, solubilisation, and mobilisation These properties permit the use of biosurfactants in the environmental field for hydrocarbon biodegradation and bioremediation. Biosurfactant applications in other industries, such as pharmaceutical, cosmetic, and food, are broadly dispersed[2,11,12,13,14]
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