Functional Approach for the Development and Production of Novel Extreme Biocatalysts

Extremophilic Oxidoreductases for the Industry: Five Successful Examples With Promising Projections

“…However, the main disadvantages of this strategy are related to the correct identification of the target enzymes, since there is currently a lack of reliable functional annotation of extremophiles genomic data, caused by the low amount of experimentally described genes. This translates in a shortfall of specific databases, and therefore a great majority of the genes from extremophiles get annotated as hypothetical proteins with unknown functions, contributing to the problem of information shortage (Caceres-Moreno et al, 2019). In addition, even if an encoding gene of interest can be identified, it might not be fully functional once expressed, and it is not yet possible through genomic methodologies to precisely determine the specific biochemical properties of an enzyme.…”

“…So far, we are currently seeing a boom of culture-independent methods for bioprospection of novel extremozymes from environmental metagenomes, which will help feed in the databases, opening the possibility to isolate and functionally characterize genes from unculturable extremophilic microorganisms (Khan and Sathya, 2018;Verma and Satyanarayana, 2020;Sysoev et al, 2021). On the other hand, there is a culture dependent functional approach for the discovery and development of novel extremozymes, which has been largely used over the years with proven successful results (Caceres-Moreno et al, 2019;Espina et al, 2020). For this, culturable extremophilic microorganisms are isolated by means of traditional microbiology techniques from environmental samples taken during scientific expeditions to extreme environments.…”

“…Subsequently, comprehensive bioinformatic analysis of the bacterial or archaeal sequenced genome allows to find the specific encoding gene of interest for its recombinant expression in a suitable heterologous host-vector system. After obtaining the recombinant extremophilic enzyme functionally overexpressed (either intra or extracellularly), it needs to be fully characterized to check the biochemical properties and compare them with the ones previously assessed in the native version (Caceres-Moreno et al, 2019).…”

“…Another solution that has proven to be efficient is taking advantage of what is already in nature by searching and discovering novel enzymes from extremophiles, as these microorganisms and their macromolecules have already adapted to thrive in environments that present extreme physicochemical conditions (e.g., high and low temperatures, acidic or alkaline pH, high pressure, salinity, among others) (Brininger et al, 2018). Consequently, extremophilic enzymes are more stable, active and robust than their mesophilic counterparts, allowing them to better perform biocatalysis under the harsh conditions found in most industrial applications (Caceres-Moreno et al, 2019). Working with extremophiles and their native enzymes, however, is not a trivial task due to the culture conditions required, their low cell yield and low enzyme expression.…”

From the Discovery of Extremozymes to an Enzymatic Product: Roadmap Based on Their Applications

“…Extremozymes have adapted to optimally perform at, or close to the environmental conditions where extremophilic microorganisms inhabit (e.g., high or low temperatures, acidic or alkaline pH, high pressure, salinity, radiation) ( Brininger et al, 2018 ). These novel extreme biocatalysts with increased stability to denaturing conditions, stand out for being highly efficient, showing higher activity, and robustness than their mesophilic counterparts, being able to carry out reactions at nonstandard conditions, producing low amounts of by-products under a wide range of extreme industrial settings ( Gomes and Steiner, 2004 ; Cáceres-Moreno et al, 2019 ).…”