Journal
BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY
Volume 63, Issue 5, Pages 625-632Publisher
WILEY-BLACKWELL
DOI: 10.1002/bab.1416
Keywords
Bacillus subtilis NX-2; poly (gamma-glutamic acid); reactive oxygen species; regulation; signal molecule; transcriptional level
Funding
- Key Projects in the National Science & Technology Pillar Program during the Twelfth Five-year Plan Period [2015BAD15B04]
- National Basic Research Program of China (973) [2013CB733603]
- National High Technology Research and Development Program of China (863) [2015AA020951]
- National Nature Science Foundation of China [21176123]
- National Key Technology RD Program [2011BAD23B04]
- Jiangsu Technology Innovation Fund for science and technology company [BC2012020]
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Effects of reactive oxygen species (ROS) on cell growth and poly(-glutamic acid) (-PGA) synthesis were studied by adding hydrogen peroxide to a medium of Bacillus subtilis NX-2. After optimizing the addition concentration and time of H2O2, a maximum concentration of 33.9g/L -PGA was obtained by adding 100 mu M H2O2 to the medium after 24H. This concentration was 20.6% higher than that of the control. The addition of diphenyleneiodonium chloride (ROS inhibitor) can interdict the effect of H2O2-induced ROS. Transcriptional levels of the cofactors and relevant genes were also determined under ROS stress to illustrate the possible metabolic mechanism contributing to the improve -PGA production. The transcriptional levels of genes belonging to the tricarboxylic acid cycle and electron transfer chain system were significantly increased by ROS, which decreased the NADH/NAD(+) ratio and increased the ATP levels, thereby providing more reducing power and energy for -PGA biosynthesis. The enhanced -PGA synthetic genes also directly promoted the formation of -PGA. This study was the first to use the ROS control strategy for -PGA fermentation and provided valuable information on the possible mechanism by which ROS regulated -PGA biosynthesis in B. subtilis NX-2. (C) 2015 International Union of Biochemistry and Molecular Biology, Inc.
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