Nucleotide Sequence and Possible Functional Mechanism of the Transcriptional Activator Gene (preL) for Neutral Protease from Lactobacillus sp.
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概要
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A positive regulatory gene (preL) from Lactobacillus sp. no. 1,which enhanced production of neutral protease (NprL), was cloned in Bacillus subtilis DB104 together with the nprL structural gene. When nprL was expressed with the regulatory gene (preL) on the same plasmid, extracellular protease production in a 12h culture was 28-fold higher (14.2U/ml) than protease production without preL expression (0.5U/ml). The nucleotide sequence of the preL was determinded and the deduced amino acid sequence revealed that PreL was relatively homologous (33%) to that of a positive regulatory protein (NprA) for protease production from Bacillus stearothermophilus, whose function is similar to that of preL. Northern hybridization analysis revealed that preL, in multicopy, enhanced transcription of the nprL gene. A consensus DNA binding motif (hylix-turn-helix) was found within the amino acid sequence of the PreL. These results suggest that preL is a transcriptional activator of pprL. Indeed, a possible target sequence of PreL with a stable palindromic structure containing putative -10 region was found in the promoter region of nprL. It is speculated that PreL melts this palindromic structure and assists RNA polymerase in binding effectively to the nprL promoter. Further-more, a similar palindromic structure was also found in the promoter region of preL itself. Enhanced production of NprL in the stationary phase may possibly be due to this auto-activation by preL.
- 社団法人日本生物工学会の論文
- 1994-04-25
著者
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TAKAGI Mutsumi
International Center for Biotechnology, Osaka University
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IMANAKA Tadayuki
Department of Synthetic and Biological Chemistry, Graduate School of Engineering, Kyoto University
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TAKAGI MASAHIRO
Department of Biotechnology, Graduate School of Engineering, Osaka University
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MAEDA Takuya
Department of Tropical Medicine and Parasitology, School of Medicine, Keio University
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Takagi M
Asahi Chemical Industry Co. Ltd. Shizuoka Jpn
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Maeda Takuya
Department Of Biotechnology Faculty Of Eigineering Osaka University
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Maeda Takuya
Department Of Biological Science And Technology Faculty Of Engineering The University Of Tokushima
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Imanaka Tadayuki
Department Of Applied Biotechnology Faculty Of Engineering Osaka University
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Takagi M
Japan Advanced Inst. Sci. And Technol. Ishikawa Jpn
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Kawano S
Kaneka Corp. Hyogo Jpn
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Kawano Shigeru
Department Of Biotechnology Faculty Of Eigineering Osaka University
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Kawano Shigeru
Fine Chemicals Research Laboratories Kaneka Corporation
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Takagi Masahiro
Department Of Biotechnology Faculty Of Engineering Osaka University
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KAWANO Shigeru
Frontier Biochemical and Medical Research Laboratories, Kaneka Corporation
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Maeda Takuya
Department of Agricultural Chemistry, University of Osaka Prefecture
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