Construction of Co-expressed MntH and Mn-catalase Gene in Escherichia coli and Optimization of Fermentation Conditions

doi:10.13560/j.cnki.biotech.bull.1985.2015.09.026

Abstract

Abstract: Mn-catalase from Thermus thermophilus HB27 and Mn²⁺ transport protein MntH from Escherichia coli were co-expressed in E. coli BL21(DE3). The optimization of fermentation medium and environment for the production of Mn-catalase was carried out at the shake flask level. The optimal carbon and nitrogen source were 7.0 g/L glycerine, 3.75 g/L yeast extract and 11.25 g/L peptone respectively. The optimum induced concentration of IPTG was 0.05 mmol/L while the Mn²⁺ in media was 1 mmol/L. In addition, the optimal initial pH of the medium and culture temperature were pH 8.0 and 37℃ respectively. Under the optimal conditions, the maximal activity of catalase reached 476 U/mL, which was 3-fold of the control. Finally, in a 5 L fermentor the activity of catalase increased to 1 094 U/mL.

Key words: Mn-catalase, transport protein, co-expression, optimization of fermentation

Wang Hui, Cui Yunfeng, Liu Yan, Shi Jiping, Zhao Zhijun, Wang Shaoming. Construction of Co-expressed MntH and Mn-catalase Gene in Escherichia coli and Optimization of Fermentation Conditions[J]. Biotechnology Bulletin, 2015, 31(9): 183-189.

References

[1] 张东旭, 堵国成, 陈坚. 微生物过氧化氢酶的发酵生产及其在纺织工业的应用[J]. 生物工程学报, 2010, 26(11)：1473-1481.
[2] 冷晒祥, 钱国坻, 华兆哲, 等. 漂白残液酶解处理对活性染料染色的影响[J]. 纺织学报, 2007, 28(7)：77-81.
[3] Chelikani P, Fita I, Loewen P. Diversity of structures and properties among catalases[J]. Cellular and Molecular Life Sciences, 2004, 61(2)：192-208.
[4] Hidalgo A, Betancor L, Mateo C, et al. Purification of a catalase from Thermus thermophilus via IMAC chromatography：effect of the support[J]. Biotechnology Progress, 2004, 20(5)：1578-1582.
[5] Thompson VS, Schaller KD, Apel WA. Purification and characterization of a novel thermo-alkali-stable catalase from Thermus brockianus[J]. Biotechnology Progress, 2003, 19(4)：1292-1299.
[6] Whittaker JW. Non-heme manganese catalase-the ‘other’ catalase[J]. Archives of Biochemistry and Biophysics, 2012, (525)：111-120.
[7] Ebara S, Shigemori Y. Alkali-tolerant high-activity catalase from a Thermophilic bacterium and its overexpression in Escherichia coli[J]. Protein Expression and Purification, 2008, 57(2)：255-260.
[8] Hidalgo A, Betancor L, Moreno R, et al. Thermus thermophilus as a cell factory for the production of a thermophilic Mn-dependent catalase which fails to be synthesized in an active form in Escherichia coli[J]. Applied and Environmental Microbiology, 2004, 70(7)：3839-3844.
[9] 何晓娟, 薛正莲, 赵志军, 等. Thermus thermophilus HB27锰过氧化氢酶在大肠杆菌中的表达及酶学性质分析[J]. 中国生物工程杂志, 2014, 34(2)：52-58.
[10] 赵志军, 华兆哲, 刘登如, 等. 碱性过氧化氢酶高产菌的筛选, 鉴定及发酵条件优化[J]. 微生物学通报, 2007, 34(4)：667-671.
[11] Zeng HW, Cai YJ, Liao XR, et al. Production, characterization, cloning and sequence analysis of a monofunctional catalase from Serratia marcescens SYBC08[J]. Journal of Basic Microbiology, 2011, 51(2)：205-214.
[12] Zeng HW, Cai YJ, Liao XR, et al. Serratia marcescens SYBC08 catalase isolated from sludge containing hydrogen peroxide shows increased catalase production by regulation of carbon metabolism[J]. Engineering in Life Sciences, 2011, 11(1)：37-43.
[13] 周丽萍, 过氧化氢酶基因工程菌的构建与发酵优化[D]. 无锡：江南大学, 2011.

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