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

Applied and Environmental Microbiology
Aurelio HidalgoJosé Berenguer

Abstract

Thermostable Mn-dependent catalases are promising enzymes in biotechnological applications as H(2)O(2)-detoxifying systems. We cloned the genes encoding Mn-dependent catalases from Thermus thermophilus HB27 and HB8 and a less thermostable mutant carrying two amino acid replacements (M129V and E293G). When the wild-type and mutant genes were overexpressed in Escherichia coli, unmodified or six-His-tagged proteins of the expected size were overproduced as inactive proteins. Several attempts to obtain active forms or to activate the overproduced proteins were unsuccessful, even when soluble and thermostable proteins were used. Therefore, a requirement for a Thermus-specific activation factor was suggested. To overcome this problem, the Mn-dependent catalase genes were overexpressed directly in T. thermophilus under the control of the Pnar promoter. This promoter belongs to a respiratory nitrate reductase from of T. thermophilus HB8, whose transcription is activated by the combined action of nitrate and anoxia. Upon induction in T. thermophilus HB8, a 20- to 30-fold increase in catalase specific activity was observed, whereas a 90- to 110-fold increase was detected when the laboratory strain T. thermophilus HB27::nar was used as th...Continue Reading

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Citations

Mar 19, 2009·Archives of Environmental Contamination and Toxicology·Tiejuan ShaoHang Min
May 8, 2008·Applied Microbiology and Biotechnology·Margarita DíazRamón Santamaría
Nov 25, 2006·Extremophiles : Life Under Extreme Conditions·Snaedis H BjornsdottirGudmundur Eggertsson
Jan 22, 2009·Extremophiles : Life Under Extreme Conditions·Felipe CavaJosé Berenguer
Jun 25, 2013·Applied and Environmental Microbiology·Hirokazu SuzukiToshihisa Ohshima
Jan 11, 2005·Applied and Environmental Microbiology·Renata MorenoJosé Berenguer
Sep 28, 2014·Biotechnology Advances·Balwinder Singh SoochMunish Puri
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Jun 19, 2012·Journal of Hazardous Materials·Ying ZhangXiaonan Dong
Jun 26, 2012·FEMS Microbiology Letters·Ying ZhangYang Wang
Aug 21, 2019·Nature Communications·Torbjørn Ølshøj JensenAlex Toftgaard Nielsen
Aug 9, 2019·Extremophiles : Life Under Extreme Conditions·Abeera ShaeerNaeem Rashid
Feb 17, 2017·Wiley Interdisciplinary Reviews. Systems Biology and Medicine·James A CountsRobert M Kelly

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