Total chemical synthesis of a thermostable enzyme capable of polymerase chain reaction | Zendy

Weiliang Xu | Zendy; Wenjun Jiang | Zendy; Jiaxing Wang | Zendy; Linping Yu | Zendy; Chen Ji | Zendy; XianYu Liu | Zendy; Lei Liu | Zendy; Ting Zhu | Zendy

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Total chemical synthesis of a thermostable enzyme capable of polymerase chain reaction

Author(s) -

Weiliang Xu,

Wenjun Jiang,

Jiaxing Wang,

Linping Yu,

Chen Ji,

XianYu Liu,

Lei Liu,

Ting Zhu

Publication year - 2017

Publication title -

cell discovery

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 3.412

H-Index - 29

ISSN - 2056-5968

DOI - 10.1038/celldisc.2017.8

Subject(s) - sulfolobus solfataricus , polymerase , polymerase chain reaction , enzyme , synthetic biology , polymerase chain reaction optimization , biochemistry , dna , chemistry , dna polymerase , microbiology and biotechnology , combinatorial chemistry , biology , computational biology , gene , nested polymerase chain reaction , archaea

Polymerase chain reaction (PCR) has been a defining tool in modern biology. Towards realizing mirror-image PCR, we have designed and chemically synthesized a mutant version of the 352-residue thermostable Sulfolobus solfataricus P2 DNA polymerase IV with l -amino acids and tested its PCR activity biochemically. To the best of our knowledge, this enzyme is the largest chemically synthesized protein reported to date. We show that with optimization of PCR conditions, the fully synthetic polymerase is capable of amplifying template sequences of up to 1.5 kb. The establishment of this synthetic route for chemically synthesizing DNA polymerase IV is a stepping stone towards building a d -enzyme system for mirror-image PCR, which may open up an avenue for the creation of many mirror-image molecular tools such as mirror-image systematic evolution of ligands by exponential enrichment.

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