Overexpression and characterization of a bacterial α-N-acetylgalactosaminidase

LIN Yi-gang; WANGYu-meng; XIA Gang

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Dec. 2014

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LIN Yi-gang, WANGYu-meng, XIA Gang. Overexpression and characterization of a bacterial α-N-acetylgalactosaminidase[J]. Journal of East China Normal University (Natural Sciences), 2012, (4): 67-74.

Citation:

LIN Yi-gang, WANGYu-meng, XIA Gang. Overexpression and characterization of a bacterial α-N-acetylgalactosaminidase[J]. Journal of East China Normal University (Natural Sciences), 2012, (4): 67-74.

LIN Yi-gang, WANGYu-meng, XIA Gang. Overexpression and characterization of a bacterial α-N-acetylgalactosaminidase[J]. Journal of East China Normal University (Natural Sciences), 2012, (4): 67-74.

Citation:

LIN Yi-gang, WANGYu-meng, XIA Gang. Overexpression and characterization of a bacterial α-N-acetylgalactosaminidase[J]. Journal of East China Normal University (Natural Sciences), 2012, (4): 67-74.

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Overexpression and characterization of a bacterial α-N-acetylgalactosaminidase

1.
Key Laboratory of Brain Functional Genomics, Ministry of Education, Shanghai Key Laboratory of Brain Functional Genomics, East China Normal University, Shanghai 200062, China

Received Date: 2011-05-01
Rev Recd Date: 2011-08-01
Publish Date: 2012-07-25

Abstract

Abstract

The coding sequence of alpha-N-acetylgalactosaminidase (A4) was amplified from genomic DNA of Chryseobacterium meningosepticum and subcloned into pET24a, which was then transformed into BL21(DE3) for overexpression of His-A4. The overexpressed His-A4 enzyme was purified by using affinity chromatography and its activity was comparable to that previously reported by using a conventional method with an artificial substrate. To better measure the activity of -N-acetylgalactosaminidase in real application, we established a novel method in which we directly used the surface antigen of red blood cell as substrate and applied ELISA to the detection of un-cleaved antigen. The activity of His-A4 was evaluated in the new ELISA method and was demonstrated to be able to decrease the blood cell surface antigen-antibody reaction in concentration- and time-dependent manner.
- α-N-acetylgalactosaminidase,
- recombination protein,
- protein purification,
- ELISA method

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References(1)

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