BIOPROSPECTION OF LIMONENE-RESISTANT MICRO-ORGANISMS AND MOLECULAR CHARACTERIZATION

Filipe Augusto Teixeira; Gustavo Molina; Tiago Antônio de Oliveira Mendes

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Paper Title

BIOPROSPECTION OF LIMONENE-RESISTANT MICRO-ORGANISMS AND MOLECULAR CHARACTERIZATION

Authors

Filipe Augusto Teixeira
Gustavo Molina
Tiago Antônio de Oliveira Mendes

Modality

Poster

Subject area

DNA and Genomics

Publishing Date

08/11/2023

Country of Publishing

Brazil | Brasil

Language of Publishing

Inglês

Paper Page

https://www.even3.com.br/anais/xmeeting2023/643252-bioprospection-of-limonene-resistant-micro-organisms-and-molecular-characterization

ISBN

978-65-272-0061-1

Keywords

Bioprospection, Limonene, Bacillus.

Summary

The food and flavoring industries have been looking for alternative methods for obtaining raw materials to reduce their reliance on the traditional methods of natural extraction and chemical synthesis of compounds. The growing demand for products with lower environmental impacts has driven this change. In this context, the biotechnological processes stand out due to the use of mild conditions, high production capacity, and generation of less waste. Among the substrates with high potential for applicability, there is limonene, a cyclic monoterpene that is the main chemical component found in citrus essential oils. Limonene is obtained at low cost and in great abundance as an industrial by-product. Despite presenting chemical and commercial characteristics favorable for biotechnological processes, limonene is toxic for most microorganisms. However, it is possible to overcome this property through a biological chassis that presents a natural phenotype of resistance to the toxicity inherent of terpenes by bioprospecting, a very effective and cheap tool for discovering new characteristics or biological functions that can be applied to industrial processes. This study aimed to prospect and identify microorganisms resistant to limonene from plant sources in Brazilian natural environments using universal genetic markers. Using enrichment culture in a selective medium containing 2% (v/v) of limonene, we were able to select 29 representatives that exhibited a terpene tolerance profile. These microorganisms were isolated from Abiu fruits (Pouteria caimito), from the subshrub Polygala fimbriata and the fruit from Solanum lycocarpum. The identification was performed by analyzing the similarity between the constitutive gene amplicons sequenced from each isolated microorganism and the NCBI (National Center for Biotechnology) database using the BLAST algorithm (Basic Local Alignment Search Tool). These results allowed the identification of six isolates. Among them, one yeast was identified at the species level as Meyerozyma caribbica, and the five other individuals belonged to the genus Bacillus. Additionally, a phylogenetic tree was generated by aligning 223 16S rRNA sequences from various representatives of Bacillus, the isolates mentioned before and Escherichia coli as an outgroup, and the results corroborated our previous findings. Regarding the bacterial representatives, two were identified at the species level as B. cereus and B. mycoides. To investigate whether resistance to limonene could be a factor common to the genus Bacillus, two representatives of the isolates in this work and three other species of this taxon (B. amyloliquefaciens, B. subtilis and B. thuringiensis) were submitted to cultivation in the presence of terpene and a trend of bacteriostatic effect was observed from concentrations above 4% (v/v) for most microorganisms tested. The genus Bacillus is widely used in industrial processes. For instance, the species B. subtilis is recognized as a universal cell factory for the production of enzymes. Our findings reinforce the importance of this taxon for biotechnology and provide possible alternatives beyond the commonly used species.

Title of the Event: X-Meeting / BSB 2023
City of the Event: Curitiba
Title of the Proceedings of the event: X-Meeting presentations
Name of the Publisher: Even3
Means of Dissemination: Meio Digital

How to cite

TEIXEIRA, Filipe Augusto; MOLINA, Gustavo; MENDES, Tiago Antônio de Oliveira. BIOPROSPECTION OF LIMONENE-RESISTANT MICRO-ORGANISMS AND MOLECULAR CHARACTERIZATION.. In: X-Meeting presentations. Anais...Curitiba(PR) Campus da indústria, 2023. Available in: https//www.even3.com.br/anais/xmeeting2023/643252-BIOPROSPECTION-OF-LIMONENE-RESISTANT-MICRO-ORGANISMS-AND-MOLECULAR-CHARACTERIZATION. Access in: 13/06/2025