Expression and Characterization of ArgR, An Arginine Regulatory Protein in Corynebacterium crenatum

CHEN Xue Lan ZHANG Bin TANG Li JIAO Hai Tao XU Heng Yi XU Feng XU Hong WEI Hua XIONG Yong Hua

CHEN Xue Lan, ZHANG Bin, TANG Li, JIAO Hai Tao, XU Heng Yi, XU Feng, XU Hong, WEI Hua, XIONG Yong Hua. Expression and Characterization of ArgR, An Arginine Regulatory Protein in Corynebacterium crenatum[J]. Biomedical and Environmental Sciences, 2014, 27(6): 436-443. doi: 10.3967/bes2014.072
Citation: CHEN Xue Lan, ZHANG Bin, TANG Li, JIAO Hai Tao, XU Heng Yi, XU Feng, XU Hong, WEI Hua, XIONG Yong Hua. Expression and Characterization of ArgR, An Arginine Regulatory Protein in Corynebacterium crenatum[J]. Biomedical and Environmental Sciences, 2014, 27(6): 436-443. doi: 10.3967/bes2014.072

doi: 10.3967/bes2014.072
基金项目: Natural Science Foundation of China, No.1360219 and No.30960012

Expression and Characterization of ArgR, An Arginine Regulatory Protein in Corynebacterium crenatum

Funds: Natural Science Foundation of China, No.1360219 and No.30960012
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  • 刊出日期:  2014-06-20

Expression and Characterization of ArgR, An Arginine Regulatory Protein in Corynebacterium crenatum

doi: 10.3967/bes2014.072
    基金项目:  Natural Science Foundation of China, No.1360219 and No.30960012

摘要: Objective Corynebacterium crenatum MT, a mutant from C. crenatum AS 1.542 with a lethal argR gene, exhibits high arginine production. To confirm the effect of ArgR on arginine biosynthesis in C. crenatum, an intact argR gene from wild-type AS 1.542 was introduced into C. crenatum MT, resulting in C. crenatum MT. sp, and the changes of transcriptional levels of the arginine biosynthetic genes and arginine production were compared between the mutant strain and the recombinant strain.
Methods Quantitative real-time polymerase chain reaction was employed to analyze the changes of the related genes at the transcriptional level, electrophoretic mobility shift assays were used to determine ArgR binding with the argCJBDF, argGH, and carAB promoter regions, and arginine production was determined with an automated amino acid analyzer.
Results Arginine production assays showed a 69.9%reduction in arginine from 9.01±0.22 mg/mL in C. crenatum MT to 2.71±0.13 mg/mL (P<0.05) in C. crenatum MT. sp. The argC, argB, argD, argF, argJ, argG, and carA genes were down-regulated significantly in C. crenatum MT. sp compared with those in its parental C. crenatum MT strain. The electrophoretic mobility shift assays showed that the promoter regions were directly bound to the ArgR protein.
Conclusion The arginine biosynthetic genes in C. crenatum are clearly controlled by the negative regulator ArgR, and intact ArgR in C. crenatum MT results in a significant descrease in arginine production.

English Abstract

CHEN Xue Lan, ZHANG Bin, TANG Li, JIAO Hai Tao, XU Heng Yi, XU Feng, XU Hong, WEI Hua, XIONG Yong Hua. Expression and Characterization of ArgR, An Arginine Regulatory Protein in Corynebacterium crenatum[J]. Biomedical and Environmental Sciences, 2014, 27(6): 436-443. doi: 10.3967/bes2014.072
Citation: CHEN Xue Lan, ZHANG Bin, TANG Li, JIAO Hai Tao, XU Heng Yi, XU Feng, XU Hong, WEI Hua, XIONG Yong Hua. Expression and Characterization of ArgR, An Arginine Regulatory Protein in Corynebacterium crenatum[J]. Biomedical and Environmental Sciences, 2014, 27(6): 436-443. doi: 10.3967/bes2014.072

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