Iaklichkin S Iu, Zimina M S, Iurchenko Iu V, Khromov I S, Neumyvakin L V
Center of Thermobiotechnology and Molecular Diagnosis, Russian Academy of Sciences, Moscow, Russia.
Genetika. 1999 Jan;35(1):37-45.
The proA proline biosynthesis gene of thermophilic bacterium Thermus ruber was cloned and sequenced, and several properties of the encoded enzyme, gamma-glutamylphosphate reductase (GPR) were studied. The proA open reading frame (ORF) was of 1286 bp. Nucleotide sequence analysis revealed the ATG initiation codon in position 36 and the TTA termination codon in position 1304. A deduced protein product of the gene was shown to be of 44,919 Da in molecular weight. The GC content was 66%, as is characteristic of various bacteria of the genus Thermus. An amino acid sequence encoded by the cloned gene showed the highest homology (up to 64%) with GPR of T. thermophilus. The maximum activity of GPR (8.2 x 10(-2) units/ml) was observed at 55 degrees C. A weak enzymatic activity was also detected at 70 degrees C. The enzyme can be used in biotechnological studies.
克隆并测序了嗜热细菌红栖热袍菌(Thermus ruber)的脯氨酸生物合成基因proA,并对编码的γ-谷氨酰磷酸还原酶(GPR)的几种特性进行了研究。proA开放阅读框(ORF)为1286 bp。核苷酸序列分析显示,起始密码子ATG位于第36位,终止密码子TTA位于第1304位。该基因推导的蛋白质产物分子量为44,919 Da。GC含量为66%,这是嗜热栖热菌属各种细菌的特征。克隆基因编码的氨基酸序列与嗜热栖热菌的GPR具有最高同源性(高达64%)。GPR的最大活性(8.2×10⁻²单位/ml)在55℃时观察到。在70℃时也检测到较弱的酶活性。该酶可用于生物技术研究。