Isolation and Identification of Thermophilic Protease Bacterium 0701 and its Protease Characterization

Abstract:

Soil was collected from Huguangyan of Zhanjiang city and regarded as separate material, It was screened thermophilic protease-producing bacteria though dilution flat, transparent circle and Folin method. Strain 0701 isolated was identificated by its morphological features, physiological and biochemical characteristics and 16s rRNA. This experiment also researched on the genetic stability of enzyme producing, and different temperature, pH influence on proteases activity. 25 high temperature-tolerance protease producing strains were isolated, which accounted for 28.1 % of all the isolated strains. Among them, strain 0701 proteases activity was 247 U/mL and it was far higher than that of proteases for other strains and the most optimum reaction temperature and pH of the proteases were 70oC and pH 7.0, respectively, and retained high enzyme activity within 50-80 oC, pH6.5-9.0. The strain 0703 still had the ability to produce protease after consecutive 4 generation, and was identified as Bacillus subtilis by morphological, physiological-biochemical characteristics and 16s rRNA.

• Periodical: Advanced Materials Research (Volumes 781-784)
• Pages: 1032-1036
• DOI: https://doi.org/10.4028/www.scientific.net/AMR.781-784.1032
• Online since: September 2013
• Authors: Ying Liu, Jun Ying An, Xu Qiong Hu, Chun Hou Xu
• Keywords: Identification, Isolation, Protease Characterization, Thermophilic Proteases Bacteria

References

[1] Carolina W M, Hamilton C, Eleni G, et al. Food Chemistry. 104-1(2007), pp.127-131.

[2] G.R. Huang, P. Huo, D.H. Dai, et a1. Journal of China Jiliang University. 18-2(2007), pp.95-98. In Chinese.

[3] C.C. Zheng, L. Zhou. Journal of Environmental Science. 32-3(2012), pp.577-583. In Chinese.

[4] B. Tang, L.F. Zhou, X.D. Chen, et a1. Acta Microbiologica Sinica. 40-2(2000), pp.155-190. In Chinese.

[5] X.C. Jia, H.G. Chen. Enzyme preparation technology. Beijing: Chemical industry press(2008). In Chinese.

[6] Yukio T, Yoshimi U. Biosei. Biotechnol. Bioehem. 62(1998), pp.1028-1030.

[7] Pavlina D, Georgieva D N. Bioehim. Biophys. Acta. 1382-2(1998), pp.207-216.

[8] Odibo F J, Obi S K. Discov. Innov. 7(1995), pp.11-14.

[9] Lu S F, chang P P. Lett. Appl. Microbiol. 22(1996), pp.5-9.

[10] Z.F. Guo, J.Q. Liu, G.Q. Chu, et a1. Quaternary scienes. 22-3(2002),P. 266-272. In Chinese.

[11] Q. Liu , Z.Y. Gu, J.Q. Liu, et a1. Marine geology & quaternary geology. 25-2(2005), P. 115-125. In Chinese.

[12] Y. Ding, J.C. Jian, Z.H. Wu, et al. Oceanologia et limnologia sinica. 40-3(2009),P. 380-384. In Chinese.

[13] QB/T 1803-1993, General methods of test for industrial enzymes. Beijing, China standard press(1994). In Chinese.

[14] X.Z. Dong, M.Y. Chai. Manual of Common Indentification for Bacteria. Beijing, Science Press(2001). In Chinese.

[15] Meera V, Saramma A V. Process Biochemistry. 41-6(2006), pp.1239-1243.

[16] D. Su. Isolation, identification and products characterization of strain SD-142 producing thermostable alkaline proteasea (2004). In Chinese.

[17] G.R. Huang. Screening and characterization of a thermostable neutral proteasea from thermophilic Bacillus sp. HS08 (2008). In Chinese.

[18] H.Y. Ma. Study on Screening and Fermentation Conditions of High-yield Neutral Protease Strains(2008). In Chinese.

[19] Y.X. Zhang, Y. Zhao, J.L. Xu, et a1. Chin J Appl Environ Biol. 13-4(2007),P. 561-564. In Chinese.