AJMB  Vol.3 No.2 , April 2013
Breeding of NHase hyper-producing Rhodococcus ruber strain LUV30-06 and verification of mutants by RAPD
Abstract: A nitrile hydratase (NHase) hyper-producing Rhodococcus ruber strain LUV30-06 was bred by mutagenization on the starting strain CGMCC3090 with ultraviolet irradiation and lithium chloride. The NHase activity of the strain LUV30-06 was increased by 21.99% (3881.3 U/ml), as compared with that of R. ruber CGMCC3090 (3181.4 U/ml). The mutant strain UV30-06 has been proved genetically stable with higher NHase activity in seven successive subcultures as well as random amplified polymorphic DNA analysis (RAPD).
Cite this paper: Wang, S. , Wang, M. and Wang, Q. (2013) Breeding of NHase hyper-producing Rhodococcus ruber strain LUV30-06 and verification of mutants by RAPD. American Journal of Molecular Biology, 3, 108-114. doi: 10.4236/ajmb.2013.32015.

[1]   Prasad, S. and Bhalla, T.C. (2010) Nitrile hydratases (NHases): At the interface of academia and industry. Biotechnology Advances, 28, 725-741. doi:10.1016/j.biotechadv.2010.05.020

[2]   Wang, S.W. and Wang, M. (2011) Research process on microbial diversity of producing NHase and study on NHase. China Biotechnology, 31, 117-132.

[3]   Zhang, J.L., Wang, M., Sun, H., et al. (2009) Isolation and characterization of Rhodococcus ruber CGMCC3090 that hydrolyzes aliphatic, aromatic and heterocyclic nitriles. African Journal of Biotechnology, 8, 5467-5475.

[4]   Shimizu S, Ogawa, J. and Kataoka, M. (1997) Screening of novel microbial enzymes for the production of biologically and chemically useful compounds. Advances in Biochemical Engineering/Biotechnology, 58, 46-87. doi:10.1007/BFb0103302

[5]   Nagasawa, T. and Yamada, H. (1990) Application of nitrile converting enzymes for the production of useful compounds. Pure & Applied Chemistry, 62, 1441-1444. doi:10.1351/pac199062071441

[6]   Guiot, P., Ryan, M.A. and Scriven, E.F.V. (1996) Preparations and applications of nicotinic acid and nicotinamide. Chimica Oggi (Chemistry Today), 14, 55-57.

[7]   Soma, Y., Kashima, M., Imaizumi, A., et al. (2005) Moisturizing effects of topical nicotinamide on atopic dry skin. International Journal of Dermatology, 44, 197-202. doi:10.1111/j.1365-4632.2004.02375.x

[8]   Xin, J. and Hu, B. (1992) Nicotinamide radiosensitizing effect and its clinical application. International Journal of Radiation Medicine and Nuclear Medicine, 16, 200-203.

[9]   Guo, J.L., Shen, H. and Li, Q. (2008) In vitro effect of nicotinamide of GLC-82 human lung adenocarcinoma cells. Guangdong Medical Journal, 29, 209-211.

[10]   Zhang, L. (2004) Niacin and niacinamide production growth gratifying and demand foreground being expected period. Medicine Economic News, 12, 25

[11]   Song, Y., Zhong, L.P. and Wang, M. (2009) Effect of metal ions on Rhodococcus sp. nitrile hydratase activity. Journal of Tianjing University of Science & Technology, 24, 11-14.

[12]   Prasad, S., Raj, J. and Bhalla, T.C. (2004) Optimization of culture conditions for hyper production of nitrile hydratase of Rhodococcus rhodochrous PA-34. Indian Journal of Microbiology, 44, 251-256.

[13]   Li, X.D., Wang, M., Zheng, Y., et al. (2010) Optimization of conditions for the biotransformation of 5-cyanovaleramide from adiponitrile by Rhodococcus ruber CGMCC-3090. 4th International Conference on Bioinformatics and Biomedical Engineering (iCBBE), Chengdu, 18-20 June 2010, 1-4.

[14]   Wang, W.S., Dong, Y., Li, T., et al. (2006) The method of DNA effective and quick means for DNA extraction of Taxol producing fungi. Journal of Science of Teachers’ College and University, 1, 60-64.

[15]   Hua, J.G. and Guo, J.H. (2003) The axonomy and application Rhodococcus. Microbiology, 30, 107-111.

[16]   Kobayashi, M. and Shimizu, S. (1998) Metalloenzyme NHase: Structure, regulation and application to biotechnology. Nature Biotechnology, 16, 733-736. doi:10.1038/nbt0898-733

[17]   Zhu, T.T., Qin, Z., Sun, X.J., et al. (2009) Breeding of high-yield strain of nitrile hydratase by compound mutation. Journal of Anhui Agricultural Sciences, 37, 1397-399.

[18]   Zhou, Y.X., Wu, J.H., Li, H.M., et al. (2008) Study on mutation and breeding of nitrile hydratase producing strain and optimization of its fermentation condition. Chemistry & Bioengineering, 25, 46-49.