Cheraghi S, Akbarzade A, Farhangi A, Chiani M, Saffari Z, Ghassemi S, Rastegari H, Mehrabi M R
Department of Biochemistry, Payam-E-Noor University, Tehran, Iran.
Pak J Biol Sci. 2010 May 15;13(10):504-8. doi: 10.3923/pjbs.2010.504.508.
The aim of this study is over-expression of Meso-diaminopimelate decarboxylase enzyme (EC 4.1.1.20) and enhancement of L-lysine production rate. The C. glutamicum LysA gene which encodes a Meso-diaminopimelate decarboxylase was cloned in E. coli. The cloned gene was sequenced; it encodes a 445 amino acids protein with molecular weight of 47 kD. Expression of the LysA gene in E. coli resulted in an increase in Meso-diaminopimelate decarboxylase activity, correlated with the presence in sodium dodecyl sulfate-polyacrylamid gels of a clear protein band that corresponds to this enzyme. The induction of cloned gene by IPTG has been shown to have an inhibitory effect on cell growth due to over-expression of the cloned gene. A two fold increase in lysine production rate was observed after introduction of the cloned gene into E. coli.
本研究的目的是过量表达中-二氨基庚二酸脱羧酶（EC 4.1.1.20）并提高L-赖氨酸的生产率。编码中-二氨基庚二酸脱羧酶的谷氨酸棒杆菌LysA基因在大肠杆菌中进行了克隆。对克隆的基因进行了测序；它编码一个分子量为47 kD的445个氨基酸的蛋白质。LysA基因在大肠杆菌中的表达导致中-二氨基庚二酸脱羧酶活性增加，这与十二烷基硫酸钠-聚丙烯酰胺凝胶中对应于该酶的一条清晰蛋白带的存在相关。IPTG对克隆基因的诱导已被证明由于克隆基因的过量表达而对细胞生长具有抑制作用。将克隆基因导入大肠杆菌后，观察到赖氨酸生产率提高了两倍。
