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| Cloning of an Mn-catalase Gene from Thermus thermophilus and Its Expression in Escherichia coli |
| HE Xiao-juan1,2, XUE Zheng-lian1,2, ZHAO Zhi-jun1,2, SUN Jun-song1,2, SHI Ji-ping1,2 |
1. Biological and Chemical Engineering Department, Anhui Polytechnic University, Wuhu, 241000, China;
2. Shanghai Advanced Research Institute of Chinese Academy of Sciences, Shanghai 201210, China |
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Abstract Thermophilic alkaline catalase is an important textile enzyme. According to the codon bias of Escherichia coli, the gene encoding manganese catalase of Thermus thermophilus HB27 were optimized. The modified gene was inserted into the expression vector pET28a(+) and transformed into Escherichia coli BL21 (DE3). After cultured with 14 mmol/L Mn2+ and induced with 0.2 mmol/L IPTG for 2 hours at 42℃, the activity of catalase reached 25 U/ml in supernatant of cell disruption. The enzyme was purified by Ni affinity chromatography and its enzyme characterization was analyzed. The optimal temperature and pH of the catalase was 70℃ and pH10.0, respectively. when the enzyme was incubated at 80℃ for 2 hours, the catalase activity was not lost. After incubated at pH9.0~11.0 for 2 hours, its activity retained 90%. The results showed that the manganese catalase has a good potential for industrial development.
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Received: 25 September 2013
Published: 25 February 2014
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