Varliero, GildaRafiq, MuhammadSingh, SwatiSummerfield, AnnabelSgouridis, FotisCowan, Don A.Barker, Gary2022-08-302022-08-302021-10Gilda Varliero, Muhammad Rafiq, Swati Singh, Annabel Summerfield, Fotis Sgouridis, Don A Cowan, Gary Barker, Microbial characterisation and Cold-Adapted Predicted Protein (CAPP) database construction from the active layer of Greenland's permafrost, FEMS Microbiology Ecology, Volume 97, Issue 10, October 2021, fiab127, https://doi.org/10.1093/femsec/fiab127.0168-6496 (print)1574-6941 (online)10.1093/femsec/fiab127https://repository.up.ac.za/handle/2263/86993Permafrost represents a reservoir for the biodiscovery of cold-adapted proteins which are advantageous in industrial and medical settings. Comparisons between different thermo-adapted proteins can give important information for cold-adaptation bioengineering. We collected permafrost active layer samples from 34 points along a proglacial transect in southwest Greenland. We obtained a deep read coverage assembly (>164x) from nanopore and Illumina sequences for the purposes of i) analysing metagenomic and metatranscriptomic trends of the microbial community of this area, and ii) creating the Cold-Adapted Predicted Protein (CAPP) database. The community showed a similar taxonomic composition in all samples along the transect, with a solid permafrost-shaped community, rather than microbial trends typical of proglacial systems. We retrieved 69 high- and medium-quality metagenome-assembled clusters, 213 complete biosynthetic gene clusters and more than three million predicted proteins. The latter constitute the CAPP database that can provide cold-adapted protein sequence information for protein- and taxon-focused amino acid sequence modifications for the future bioengineering of cold-adapted enzymes. As an example, we focused on the enzyme polyphenol oxidase, and demonstrated how sequence variation information could inform its protein engineering.en© The Author(s) 2021. Published by Oxford University Press on behalf of FEMS. This is an Open Access article distributed under the terms of the Creative Commons Attribution License.Cold-adapted protein (CAPP)BioengineeringSoil metagenomicsSoil metatranscriptomicsPermafrostGreenland ice sheet (GrIS)Article