Journalartikel

Jahr der Veröffentlichung: 2022

Seiten: 5563-5574

Zeitschrift: Applied Microbiology and Biotechnology

Bandnummer: 106

Heftnummer: 17

ISSN: 0175-7598

eISSN: 1432-0614

Open Access Status: Hybrid

DOI Link: https://doi.org/10.1007/s00253-022-12071-1

Verlag: Springer

Abstract: 
Amine transaminases (ATA) convert ketones into optically active amines and are used to prepare active pharmaceutical ingredients and building blocks. Novel ATA can be identified in protein databases due to the extensive knowledge of sequence-function relationships. However, predicting thermo- and operational stability from the amino acid sequence is a persisting challenge and a vital step towards identifying efficient ATA biocatalysts for industrial applications. In this study, we performed a database mining and characterized selected putative enzymes of the beta-alanine:pyruvate transaminase cluster (3N5M) - a subfamily with so far only a few described members, whose tetrameric structure was suggested to positively affect operational stability. Four putative transaminases (TA-1: Bilophilia wadsworthia, TA-5: Halomonas elongata, TA-9: Burkholderia cepacia, and TA-10: Burkholderia multivorans) were obtained in a soluble form as tetramers in E. coli. During comparison of these tetrameric with known dimeric transaminases we found that indeed novel ATA with high operational stabilities can be identified in this protein subfamily, but we also found exceptions to the hypothesized correlation that a tetrameric assembly leads to increased stability. The discovered ATA from Burkholderia multivorans features a broad substrate specificity, including isopropylamine acceptance, is highly active (6 U/mg) in the conversion of 1-phenylethylamine with pyruvate and shows a thermostability of up to 70 degrees C under both, storage and operating conditions. In addition, 50% (v/v) of isopropanol or DMSO can be employed as co-solvents without a destabilizing effect on the enzyme during an incubation time of 16 h at 30 degrees C.

Harvard-Zitierstil: Kollipara, M., Matzel, P., Sowa, M., Brott, S., Bornscheuer, U. and Hoehne, M. (2022) Characterization of proteins from the 3N5M family reveals an operationally stable amine transaminase, Applied Microbiology and Biotechnology, 106(17), pp. 5563-5574. https://doi.org/10.1007/s00253-022-12071-1

APA-Zitierstil: Kollipara, M., Matzel, P., Sowa, M., Brott, S., Bornscheuer, U., & Hoehne, M. (2022). Characterization of proteins from the 3N5M family reveals an operationally stable amine transaminase. Applied Microbiology and Biotechnology. 106(17), 5563-5574. https://doi.org/10.1007/s00253-022-12071-1