Laboratory of Natural Products Chemistry and Biomolecules (LNPC-BioM), Faculty of Sciences, University of Blida 1, Road of Soumaâ, PO Box 270, 09000 Blida, Algeria; Laboratoire de Biologie des Systèmes Microbiens (LBSM), École Normale Supérieure (ENS) de Kouba, PO Box 92 Vieux-Kouba, 16308, Alger, Algeria; Laboratory of Microbial Biotechnology and Engineering Enzymes (LMBEE), Centre of Biotechnology of Sfax (CBS), University of Sfax, Road of Sidi Mansour Km 6, PO Box 1177, Sfax 3018, Tunisia.
Laboratory of Microbial Biotechnology and Engineering Enzymes (LMBEE), Centre of Biotechnology of Sfax (CBS), University of Sfax, Road of Sidi Mansour Km 6, PO Box 1177, Sfax 3018, Tunisia.
Int J Biol Macromol. 2018 Feb;107(Pt B):2351-2363. doi: 10.1016/j.ijbiomac.2017.10.116. Epub 2017 Oct 18.
A novel extracellular protease called MPDZ was purified and characterized from Pseudomonas fluorescens strain TBS09. The enzymatic properties of MPDZ were investigated using biochemical and biophysical methods. Matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS) analysis revealed that it was a monomer with a molecular mass of 50013.17Da. The NH-terminal 27 amino acid sequence of MPDZ showed high homology with those of Pseudomonas-proteases of the serralysin family. MPDZ showed optimal activity at pH 7 and 60°C. It was totally inhibited by EGTA, EDTA, and 1,10-phenanthroline, suggesting its belonging to the metalloprotease family. Because of the interesting properties, the mpDZ gene encoding MPDZ was cloned, sequenced, and expressed in E. coli. The deduced amino acid sequence showed a strong homology with other Pseudomonas-metalloproteases. The highest sequence identity value (97%) was obtained with AprX from P. fluorescens strain CY091, with only 12 different amino acid residues. The physico-chemical properties of the extracellular purified recombinant enzyme (rMPDZ) were similar to those of MPDZ. Overall, MPDZ is bestowed with a number of promising biochemical properties that might give new opportunities for its biocatalytic applications. These data constitute an essential first step towards an understanding of the properties of MPDZ enzyme.
一种新型细胞外蛋白酶 MPDZ 从荧光假单胞菌 TBS09 中被分离和鉴定。使用生化和生物物理方法研究了 MPDZ 的酶学特性。基质辅助激光解吸电离飞行时间质谱（MALDI-TOF/MS）分析表明它是一种分子量为 50013.17Da 的单体。MPDZ 的 NH-末端 27 个氨基酸序列与丝氨酸蛋白酶家族的假单胞菌蛋白酶具有高度同源性。MPDZ 在 pH7 和 60°C 时表现出最佳活性。它被 EGTA、EDTA 和 1,10-菲咯啉完全抑制，表明它属于金属蛋白酶家族。由于其有趣的特性，mpDZ 基因被克隆、测序并在大肠杆菌中表达。推导的氨基酸序列与其他假单胞菌金属蛋白酶具有很强的同源性。与 P. fluorescens 菌株 CY091 的 AprX 相比，具有最高的序列同一性值（97%），只有 12 个不同的氨基酸残基。细胞外纯化的重组酶（rMPDZ）的理化性质与 MPDZ 相似。总的来说，MPDZ 具有许多有前途的生化特性，这可能为其生物催化应用提供新的机会。这些数据构成了理解 MPDZ 酶特性的重要的第一步。
