Estimating the success of enzyme bioprospecting through metagenomics: current status and future trends

Author： Ferrer Manuel Martínez-Martínez Mónica Bargiela Rafael Streit Wolfgang R. Golyshina Olga V. Golyshin Peter N.

E-ISSN： 1751-7915|9|1|22-34

ISSN： 1751-7907

Source： Microbial Biotechnology, Vol.9, Iss.1, 2016-01, pp. : 22-34

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Abstract

Recent reports have suggested that the establishment of industrially relevant enzyme collections from environmental genomes has become a routine procedure. Across the studies assessed, a mean number of approximately 44 active clones were obtained in an average size of approximately 53 000 clones tested using naïve screening protocols. This number could be significantly increased in shorter times when novel metagenome enzyme sequences obtained by direct sequencing are selected and subjected to high-throughput expression for subsequent production and characterization. The pre-screening of clone libraries by naïve screens followed by the pyrosequencing of the inserts allowed for a 106-fold increase in the success rate of identifying genes encoding enzymes of interest. However, a much longer time, usually on the order of years, is needed from the time of enzyme identification to the establishment of an industrial process. If the hit frequency for the identification of enzymes performing at high turnover rates under real application conditions could be increased while still covering a high natural diversity, the very expensive and time-consuming enzyme optimization phase would likely be significantly shortened. At this point, it is important to review the current knowledge about the success of fine-tuned naïve- and sequence-based screening protocols for enzyme selection and to describe the environments worldwide that have already been subjected to enzyme screen programmes through metagenomic tools. Here, we provide such estimations and suggest the current challenges and future actions needed before environmental enzymes can be successfully introduced into the market. Graphical image. Routine steps for the establishment of industrially relevant enzyme collections from pure cultures and environmental genomes. The figure summarizes the different steps for fine-tuned naïve- and sequence-based screening protocols for enzyme selection. Also, the further steps for production, preparation and testing of new enzymes for industrially-relevant processes, are summarized.