Biotechnology1682-296x1682-2978Asian Network for Scientific Information10.3923/biotech.2017.10.18Rhodococcus ruber OA1]]>WangZhenglong SunYing ShiYaru SongWenteng ZhangChunyang 12017161Background and Objective: Salicylic Acid (SA) is widely used in medicine and food. Rhodococcus ruber OA1 utilizes salicylate as the sole carbon and energy source for growth and catechol 1,2-dioxygenase (C12O) is detected in R. ruber OA1 grown on salicylate. However, C12O has not previously been physiologically or biochemically characterized in R. ruber. The aim of this study was to characterize C12O from R. ruber OA1 (OA1-C12O). Methodology: The catechol 1,2-dioxygenase gene (catA) from R. ruber OA1 (OA1-catA) was cloned into the pEASY-E1 vector to obtain the recombinant plasmid pEASY-E1-catA and C12O was expressed in Escherichia coli BL21 (DE3). The heterologously expressed OA1-C12O was purified and its physiological and biochemical characteristics were further studied. Results: Based on the phylogenetic analysis of catA gene, it was found that OA1-catA clustered with other catA genes from Gram-positive bacteria including Nocardia sp. C-14-1, Streptomyces ghanaensis ATCC 14672 and Amycolatopsis orientalis HCCB10007. After the expression and enzymatic characterization of C12O, it was revealed that the expressed OA1-C12O had the ability to degrade catechol to cis,cis-muconic acid with a specific enzyme of 231.4 U mg1 protein. The optimal reaction conditions of OA1-C12O were 25°C and pH 7.0. Besides, Mn2+ could increase the activity of OA1-C12O, while Mg2+ and NH4+ inhibited its activity. Conclusion: In this study the C12O from R. ruber OA1 was successfully expressed in E. coli BL21 (DE3) for the first time and its catalytic characteristics were explored in detail.]]>Vlot, A.C., D.M.A. Dempsey and D.F. Klessig,200947177206Stamatis, N.K. and I.K. Konstantinou,201348800813Evgenidou, E.N., I.K. Konstantinou and D.A. Lambropoulou,2015505905926Verenitch, S.S., C.J. Lowe and A. Mazumder,20061116193203Seo, J.S., Y.S. Keum and Q.X. Li,20096278309Uemura, T., A. Kita, Y. Watanabe, M. Adachi, R. Kuroki and Y. Morimoto,2016469158163Fang, T. and N.Y. Zhou,2014Ralstonia sp. strain U2.]]>98671679Park, J.Y., J.W. Hong and G.M. Gadd,2009Fusarium oxysporum GJ4 is affected by toxic catalytic polymerization mediated by copper oxide.]]>75765771Kandeel, M., A. Altaher, Y. Kitade, M. Abdelaziz, M. Alnazawi and K. Elshazli,201664271280Schubert, J., F.D. Muller, R. Schmidt, A. Luch and T.G. Schulz,20158921292139Mansoor S., N. Gupta, G. Luczy-Bachman, G.A. Limb, B.D. Kuppermann and M.C. Kenney,2010in vitro.]]>271107114Matera, I., M. Ferraroni, M. Kolomytseva, L. Golovleva, A. Scozzafava and F. Briganti,2010Rhodococcus opacus 1CP: quantitative structure/activity relationship and the crystal structures of native enzyme and catechols adducts.]]>170548564Guzik, U., I. Gren, K. Hupert-Kocurek and D. Wojcieszynska,2011Pseudomonas putida strain N6.]]>65504512Giedraityte, G. and L. Kalediene,2009Geobacillus sp. strain: Purification and properties.]]>46873Gou, M., Y.Y. Qu, J.T. Zhou, A. Li and M.S. Uddin,2009Sphingomonas xenophaga QYY.]]>52615620Pandeeti, E.V.P. and D. Siddavattam,2011Acinetobacter sp. DS002 and cloning, sequencing of partial catA gene.]]>51312318Lin, J. and R.N. Milase,2015Acinetobacter sp. Y64 strain and Escherichia coli transformants.]]>34421433Larkin, M.A., G. Blackshields, N.P. Brown, R. Chenna and P.A. McGettigan et al.,20072329472948Tamura, K., G. Stecher, D. Peterson, A. Filipski and S. Kumar,20133027252729Packeiser, H., C. Lim, B. Balagurunathan, J. Wu and H. Zhao,2013169695700Zhao, Y., Z. Dong, T. Li and G. Huang,2014174649Gao, X., J. Feng, Q. Hua, D. Wei and X. Wang,201432343347Ku, H.K., H.M. Lim, K.H. Oh, H.J. Yang, J.S. Jeong and S.K. Kim,2013434178180Yao, Z., C. Zhang, F. Lu, X. Bie and Z.X. 2012Issatchenkia terricola strain XJ-2.]]>9319992009Wang, L., G. Fang and D. Cao,20153+ with colorimetric and fluorometric dual-mode.]]>207849857Huo, J., K. Liu, X. Zhao, X. Zhang and Y. Wang,20142+ in aqueous buffer.]]>17789792Nadaf, N.H. and J.S. Ghosh,2011Rhodococcus sp. NCIM 2891.]]>3608613Wang, C.L., S.L. You and S.L. Wang,2006Pseudomonas aeruginosa with benzoic acid as a carbon source.]]>4115941601Caglio, R., E. Pessione, F. Valetti, C. Giunta and E. Ghibaudi,2013Acinetobacter radioresistens S13.]]>267584Long, Y., S. Yang, Z. Xie and L. Cheng,2016Candida tropicalis JH8 strain.]]>46673678Guzik, U., K. Hupert-Kocurek, M. Sitnik and D. Wojcieszynska,2013Stenotrophomonas maltophilia strain KB2 as a useful tool in cis,cis-muconic acid production.]]>10312971307