title: Discovery and characterization of a second extremely thermostable (+)-γ-lactamase from Sulfolobus solfataricus P2.
abstract: A thermostable formamidase from the hyperthermophilic archaeon Sulfolobus solfataricus P2 was revealed to be a novel, thermostable (+)-γ-lactamase. This (+)-γ-lactamase (Sso2810) is composed of only 318 amino acid residues, in contrast to a previously reported (+)-γ-lactamase (Sso2122) with 504 amino acid residues from the same strain. Herein, we demonstrate that a single strain may contain diverse (+)-γ-lactamases. The gene of this thermostable (+)-γ-lactamase was cloned, functionally expressed in Escherichia coli 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 (-)-γ-lactam for the synthesis of well-known carbocyclic nucleoside antiretroviral agents like Abacavir and Peramivir. The chiral product of the enzyme was purified to >99% enantiomeric excess.
authors.affiliation: State Key Laboratory of Chemical Resources Engineering, Beijing University of Chemical Technology, Beijing 100029, People's Republic of China.
authors.forename: Shaozhou
authors.initials: S
authors.lastname: Zhu
authors.affiliation: State Key Laboratory of Chemical Resources Engineering, Beijing University of Chemical Technology, Beijing 100029, People's Republic of China.
authors.forename: Rong
authors.initials: R
authors.lastname: Huang
authors.affiliation: State Key Laboratory of Chemical Resources Engineering, Beijing University of Chemical Technology, Beijing 100029, People's Republic of China.
authors.forename: Shuaihua
authors.initials: S
authors.lastname: Gao
authors.affiliation: State Key Laboratory of Chemical Resources Engineering, Beijing University of Chemical Technology, Beijing 100029, People's Republic of China.
authors.forename: Xinxin
authors.initials: X
authors.lastname: Li
authors.affiliation: State Key Laboratory of Chemical Resources Engineering, Beijing University of Chemical Technology, Beijing 100029, People's Republic of China. Electronic address: zhenggj@mail.buct.edu.cn.
authors.forename: Guojun
authors.initials: G
authors.lastname: Zheng
mesh_terms: D000146:Acids, Carbocyclic; D000581:Amidohydrolases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D003001:Cloning, Molecular; D003517:Cyclopentanes / Q000378:metabolism; D015224:Dideoxynucleosides / Q000378:metabolism; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D006146:Guanidines / Q000378:metabolism; D006358:Hot Temperature; D010802:Phylogeny; D048229:Sulfolobus solfataricus / Q000201:enzymology* / Q000235:genetics; D000099227:Abacavir
chemical_list: D000146:Acids, Carbocyclic; D000581:Amidohydrolases; D003517:Cyclopentanes; D015224:Dideoxynucleosides; D006146:Guanidines; D000099227:Abacavir; C101846:formamidase; C401943:gamma-lactamase; C414210:peramivir
keywords: (+)-γ-Lactamase; Enzyme promiscuity; Nucleotide analogues; Stereoselective hydrolysis; Sulfolobus solfataricus