TY  - JOUR
AU  - Ferrandi, Erica E.
AU  - Spasić, Jelena
AU  - Đokić, Lidija
AU  - Vainshtein, Yevhen
AU  - Senthamaraikannan, Ramsankar
AU  - Vojnović, Sandra
AU  - Grumaz, Christian
AU  - Monti, Daniela
AU  - Nikodinović-Runić, Jasmina
PY  - 2021
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1455
AB  - Three Streptomyces sp. strains with a multitude of target enzymatic activities confirmed by functional screening, namely BV129, BV286 and BV333, were subjected to genome sequencing aiming at the annotation of genes of interest, in-depth bioinformatics characterization and functional expression of the biocatalysts. A whole-genome shotgun sequencing followed by de novo genome assembly and annotation was performed revealing genomes of 6.4, 9.4 and 7.3 Mbp, respectively. Functional annotation of the proteins of interest resulted in between 2047 and 2763 putative targets. Among the various enzymatic activities that the three Streptomyces strains demonstrated to produce by functional screening, we focused our attention on transaminases (TAs) and laccases due to their high biocatalytic potential. Bioinformatics search allowed the identification of a putative TA from Streptomyces sp. BV333 as a potentially novel broad substrate scope TA and a putative laccase from Streptomyces sp. BV286 as potentially novel blue multicopper oxidase. The two sequences were cloned and overexpressed in Escherichia coli and the two novel enzymes, transaminase Sbv333-TA and laccase Sbv286-LAC, were characterized. Interestingly, both enzymes resulted to be exceptionally thermostable, Sbv333-TA showing a melting temperature (T-M = 85 degrees C) only slightly lower compared to the T-M of the most thermostable transaminases described to date (87-88 degrees C) and Sbv286-LAC being even thermoactivated at temperature  gt 60 degrees C. Moreover, Sbv333-TA showed a broad substrate scope and remarkably demonstrated to be active in the transamination of beta-ketoesters, which are rarely accepted by currently known TAs. On the other hand, Sbv286-LAC showed an improved activity in the presence of the cosolvent acetonitrile. Overall, it was shown that a combination of approaches from standard microbiological and biochemical screens to genome sequencing and analysis is required to afford novel and functional biocatalysts.
PB  - MDPI, Basel
T2  - Catalysts
T1  - Novel Transaminase and Laccase from Streptomyces spp. Using Combined Identification Approaches
IS  - 8
VL  - 11
DO  - 10.3390/catal11080919
ER  - 

@article{
author = "Ferrandi, Erica E. and Spasić, Jelena and Đokić, Lidija and Vainshtein, Yevhen and Senthamaraikannan, Ramsankar and Vojnović, Sandra and Grumaz, Christian and Monti, Daniela and Nikodinović-Runić, Jasmina",
year = "2021",
abstract = "Three Streptomyces sp. strains with a multitude of target enzymatic activities confirmed by functional screening, namely BV129, BV286 and BV333, were subjected to genome sequencing aiming at the annotation of genes of interest, in-depth bioinformatics characterization and functional expression of the biocatalysts. A whole-genome shotgun sequencing followed by de novo genome assembly and annotation was performed revealing genomes of 6.4, 9.4 and 7.3 Mbp, respectively. Functional annotation of the proteins of interest resulted in between 2047 and 2763 putative targets. Among the various enzymatic activities that the three Streptomyces strains demonstrated to produce by functional screening, we focused our attention on transaminases (TAs) and laccases due to their high biocatalytic potential. Bioinformatics search allowed the identification of a putative TA from Streptomyces sp. BV333 as a potentially novel broad substrate scope TA and a putative laccase from Streptomyces sp. BV286 as potentially novel blue multicopper oxidase. The two sequences were cloned and overexpressed in Escherichia coli and the two novel enzymes, transaminase Sbv333-TA and laccase Sbv286-LAC, were characterized. Interestingly, both enzymes resulted to be exceptionally thermostable, Sbv333-TA showing a melting temperature (T-M = 85 degrees C) only slightly lower compared to the T-M of the most thermostable transaminases described to date (87-88 degrees C) and Sbv286-LAC being even thermoactivated at temperature  gt 60 degrees C. Moreover, Sbv333-TA showed a broad substrate scope and remarkably demonstrated to be active in the transamination of beta-ketoesters, which are rarely accepted by currently known TAs. On the other hand, Sbv286-LAC showed an improved activity in the presence of the cosolvent acetonitrile. Overall, it was shown that a combination of approaches from standard microbiological and biochemical screens to genome sequencing and analysis is required to afford novel and functional biocatalysts.",
publisher = "MDPI, Basel",
journal = "Catalysts",
title = "Novel Transaminase and Laccase from Streptomyces spp. Using Combined Identification Approaches",
number = "8",
volume = "11",
doi = "10.3390/catal11080919"
}

Ferrandi, E. E., Spasić, J., Đokić, L., Vainshtein, Y., Senthamaraikannan, R., Vojnović, S., Grumaz, C., Monti, D.,& Nikodinović-Runić, J.. (2021). Novel Transaminase and Laccase from Streptomyces spp. Using Combined Identification Approaches. in Catalysts
MDPI, Basel., 11(8).
https://doi.org/10.3390/catal11080919

Ferrandi EE, Spasić J, Đokić L, Vainshtein Y, Senthamaraikannan R, Vojnović S, Grumaz C, Monti D, Nikodinović-Runić J. Novel Transaminase and Laccase from Streptomyces spp. Using Combined Identification Approaches. in Catalysts. 2021;11(8).
doi:10.3390/catal11080919 .

Ferrandi, Erica E., Spasić, Jelena, Đokić, Lidija, Vainshtein, Yevhen, Senthamaraikannan, Ramsankar, Vojnović, Sandra, Grumaz, Christian, Monti, Daniela, Nikodinović-Runić, Jasmina, "Novel Transaminase and Laccase from Streptomyces spp. Using Combined Identification Approaches" in Catalysts, 11, no. 8 (2021),
https://doi.org/10.3390/catal11080919 . .