A thermostable d-polymerase for mirror-image PCR.
Biological evolution resulted in a homochiral world in which nucleic acids consist exclusively of d-nucleotides and proteins made by ribosomal translation of l-amino acids. From the perspective of synthetic biology, however, particularly anabolic enzymes that could build the mirror-image counterparts of biological macromolecules such as l-DNA or l-RNA are lacking. Based on a convergent synthesis strategy, we have chemically produced and characterized a thermostable mirror-image polymerase that efficiently replicates and amplifies mirror-image (l)-DNA. This artificial enzyme, dubbed d-Dpo4-3C, is a mutant of Sulfolobus solfataricus DNA polymerase IV consisting of 352 d-amino acids. d-Dpo4-3C was reliably deployed in classical polymerase chain reactions (PCR) and it was used to assemble a first mirror-image gene coding for the protein Sso7d. We believe that this d-polymerase provides a valuable tool to further investigate the mysteries of biological (homo)chirality and to pave the way for potential novel life forms running on a mirror-image genome.
NOXXON Pharma AG, Weinbergweg 23, 06120 Halle (Saale), Germany.
Andreas
A
Pech
NOXXON Pharma AG, Max-Dohrn-Str. 8-10, 10589 Berlin, Germany.
John
J
Achenbach
NOXXON Pharma AG, Max-Dohrn-Str. 8-10, 10589 Berlin, Germany.
Michael
M
Jahnz
NOXXON Pharma AG, Weinbergweg 23, 06120 Halle (Saale), Germany.
Simone
S
Schülzchen
NOXXON Pharma AG, Max-Dohrn-Str. 8-10, 10589 Berlin, Germany.
Florian
F
Jarosch
Institute of Biochemistry and Biotechnology, Martin-Luther-University Halle-Wittenberg, Kurt-Mothes-Strasse 3, 06120 Halle (Saale), Germany.
Frank
F
Bordusa
NOXXON Pharma AG, Max-Dohrn-Str. 8-10, 10589 Berlin, Germany.
Sven
S
Klussmann
D019843:Archaeal Proteins / Q000235:genetics; D004247:DNA / Q000096:biosynthesis* / Q000737:chemistry; D019951:DNA Polymerase beta / Q000138:chemical synthesis / Q000235:genetics* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics; D004795:Enzyme Stability; D009154:Mutation; D013237:Stereoisomerism; D048229:Sulfolobus solfataricus / Q000201:enzymology; D013696:Temperature
D019843:Archaeal Proteins; D004268:DNA-Binding Proteins; C093228:Sso7d protein, Sulfolobus; D004247:DNA; D019951:DNA Polymerase beta