Discovery and characterization of a second extremely thermostable (+)-γ-lactamase from Sulfolobus solfataricus P2

作者：Zhu, Shaozhou; Huang, Rong; Gao, Shuaihua; Li, Xinxin; Zheng, Guojun^*

来源：Journal of Bioscience and Bioengineering, 2016, 121(5): 484-490.

DOI：10.1016/j.jbiosc.2015.09.012

• Sulfolobus solfataricus
• (+)-gamma-Lactamase
• Stereoselective hydrolysis
• Enzyme promiscuity
• Nucleotide analogues

A thermostable formamidase from the hyperthermophilic archaeon Sulfolobus solfataricus P2 was revealed to be a novel, thermostable ( +)-gamma-lactamase. This ( +)-gamma-lactamase (Sso2810) is composed of only 318 amino acid residues, in contrast to a previously reported ( +)-gamma-lactamase (Sso2122) with 504 amino acid residues from the same strain. Herein, we demonstrate that a single strain may contain diverse ( +)-gamma-lactamases. The gene of this thermostable ( +)-gamma-lactamase was cloned, functionally expressed in Escherichia coil BL21 and purified by a simple yet effective heat treatment method. Sso2810 was biochemically characterized and compared to Sso2122, with phylogenetic analysis indicating different evolutionary histories for the two encoding genes. This newly found thermostable enzyme shows promising properties for industrial applications; specifically, it could be used for the production of chirally pure (-)-gamma-lactam for the synthesis of well-known carbocyclic nucleoside antiretroviratakents like Abacavir and Peramivir. The chiral product of the enzyme was purified to >99% enantiomeric excess.

• 出版日期2016-5
• 单位北京化工大学