GENOME OF DIUTINA RUGOSA WITH BIOTECHNOLOGICAL POTENTIAL - IDENTIFICATION OF HYDROLASES

Bernardo Saraiva Velozo; Priscila E. de Faria; Rodrigo Volcan Almeida; Denise Maria Guimarães Freire; Elisa dAvila Costa Cavalcanti; Rafael Dias Mesquita

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Paper Title

GENOME OF DIUTINA RUGOSA WITH BIOTECHNOLOGICAL POTENTIAL - IDENTIFICATION OF HYDROLASES

Authors

Bernardo Saraiva Velozo
Priscila E. de Faria
Rodrigo Volcan Almeida
Denise Maria Guimarães Freire
Elisa dAvila Costa Cavalcanti
Rafael Dias Mesquita

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/xmeeting2023/638373-genome-of-diutina-rugosa-with-biotechnological-potential---identification-of-hydrolases

ISBN

978-65-272-0061-1

Keywords

Genomics, Bioinformatics, Diutina rugosa, Biotechnology

Summary

Exploring enzymes has become increasingly significant due to their potential to produce more eco-friendly and cost-effective alternatives to conventional chemical processes, which in turn could enhance competitiveness and sustainability.The AB_hydrolase clan includes enzymes that are widely used in the food, pharmaceutical, cosmetic, and biofuel industries. Moreover, enzymes from Diutina rugosa, which are commercially available in the aforementioned industries, hold great potential for biotechnological applications in other areas. However, due to limited genetic information available, sequencing is necessary to uncover the biotechnological diversity of strains from different origins. Therefore, our objective is to sequence and assemble the genome of the D. rugosa ATCC-14830 strain, from industrial origin, predict genes and identify enzymes belonging to the AB_hydrolase clan. To identify hydrolases in the genome of the reference strain CBS613, we selected enzymes with conserved domains belonging to the 75 families that constitute the AB_hydrolase clan (CL0028). We performed a mechanical cell lysis step with liquid nitrogen, followed by genomic DNA extraction using the Genomic DNA Purification kit (FERMENTAS). Subsequently, sequencing was conducted on the Illumina platform, using paired reads of 150 bp. We ensured quality control with FastQC and conducted read cleaning using AdapterRemoval. Genome assembly was executed with the Spades program, and the assembly quality was evaluated using QUAST and BUSCO to search for orthologs in nearby species. Gene prediction will be conducted using the FunGAP program. After gene prediction, we will apply the same selection criteria used for the CBS613 strain. Additionally, a comparison between the strains will be conducted to analyze the hydrolase content. We identified eighty hydrolases in the genome of the CBS613 strain, categorized into the families of proteases (30), lipases (13), esterases (11), hydrolases (10), thioesterases (3), transferases (1), polymerases (0), and tannases (0). The sequencing of the ATCC-14830 strain genome generated 41,090,998 reads, and the cleaning process involved filtering the adapters (TruSeq3 kit), removing regions with quality below 30, and a minimum size of 70 bp. These parameters resulted in a loss of only 0.859% of the reads. The assembly has a total size of 4,309,873 bp, distributed in 352 contigs, yielding a 4 Mbp genome, with BUSCO showing a total of 6% completeness. The results revealed a number of hydrolases in the reference strain that are compatible with other organisms of the same genus. Regarding the analysis of the ATCC-14830 strain genome, all sequencing quality parameters met our expectations. However, when analyzing the genome size, we found it to be smaller than expected, with a size of 4 Mbp, which is significantly smaller than the expected size of 13 Mbp for the CBS613 strain genome. Consequently, our next step will involve another extraction and sequencing of the DNA from the ATCC-14830 strain using the MinION sequencer. We will combine the sequences and repeat the assembly and genomic quality analysis steps.

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

VELOZO, Bernardo Saraiva et al.. GENOME OF DIUTINA RUGOSA WITH BIOTECHNOLOGICAL POTENTIAL - IDENTIFICATION OF HYDROLASES.. In: X-Meeting presentations. Anais...Curitiba(PR) Campus da indústria, 2023. Available in: https//www.even3.com.br/anais/xmeeting2023/638373-GENOME-OF-DIUTINA-RUGOSA-WITH-BIOTECHNOLOGICAL-POTENTIAL---IDENTIFICATION-OF-HYDROLASES. Access in: 20/05/2025