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  o-Nitrophenyl-β-d-galactopyranoside 6-phosphate

Hengstenberg, W., & Morse, M. L. (1975). o-Nitrophenyl-β-d-galactopyranoside 6-phosphate. Methods in Enzymology, 41, 119-120. doi:10.1016/S0076-6879(75)41030-8.

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MethEnzymol_41_1975_119.pdf (Any fulltext), 931KB
 
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Hengstenberg, Wolfgang1, Author           
Morse, M. L.1, Author           
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1Max Planck Institute for Medical Research, Max Planck Society, ou_1125545              

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 Abstract: This chapter discusses o-nitrophenyl-β-D-galactopyranoside 6-phosphate. The procedure for phosphorylating primary hydroxyl groups in unprotected nucleosides is applied to o-nitrophenyl- β -D-galactoside (ONPG) to synthesize the 6-phosphate. Phosphorus oxychloride in trimethyl phosphate containing a small amount of water yielded ONPG-6-P in 50% yield. The method should be applicable to the synthesis of the 6-phosphates of other ONP glycosides. The procedure given is much simpler than that employed initially to synthesize ONPG 6-P: 30 mmoles of o-nitrophenyl β-D-galactoside is dissolved in 75 ml of trimethyl phosphate containing 30 mmoles of water and 90 mmoles of phosphorus oxychloride. The mixture is stirred and kept for 3 hr in an ice bath. Crushed ice is then added, Phosphoric and hydrochloric acids are neutralized with concentrated ammonia, and the yellow solution is evaporated under diminished pressure, o-Nitrophenol is removed by evaporating water several times from the solution. The colorless solution contained 14.65 mmoles of ONPG 6-P as determined enzymically. Enzymic determination of the ONPG 6-phosphate is performed with 10 μg of a highly purified, electrophoretically homogeneous, enzyme preparation in 2.5 ml of the buffer. For high voltage paper electrophoresis, 3 mg of ONPG-6P as the dicyclohexylammonium salt, is hydrolyzed in the standard buffer with 10 μg of 6-phosphogalactosidase.

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Language(s): eng - English
 Dates: 2003-12-161975
 Publication Status: Published in print
 Pages: 2
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 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1016/S0076-6879(75)41030-8
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Title: Methods in Enzymology
  Other : Methods Enzymol.
Source Genre: Journal
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Publ. Info: New York, NY : Academic Press
Pages: - Volume / Issue: 41 Sequence Number: - Start / End Page: 119 - 120 Identifier: ISSN: 0076-6879
CoNE: https://pure.mpg.de/cone/journals/resource/110975506069301