Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.7.10.1 (ERK)
 95,504 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Calcium 2-keto-L-gulonate (Ca-2-KLG, a key intermediate in vitamin C synthesis) is produced from calcium 2,5-diketo D-gluconate (Ca-2,5-DKG) by a variety of bacteria. A few bacterial species which efficiently convert glucose to Ca-2,5-DKG have been isolated in our laboratory. Our bacterial collection included species that possess the genes for production of Ca-2-KLG from Ca-2,5-DKG; however, the yield of the former is poor. A procedure for the preparation of spheroplasts in Ca-2,5-DKG- and Ca-2-KLG-producing bacteria was developed for the construction of recombinants (fusants), combining the genes for conversion of glucose to Ca-2-KLG efficiently by protoplast fusion. The standard procedure for spheroplast formation in Gram negative bacteria by the Tris-sucrose-EDTA-lysozyme system did not work in the organisms under investigation. The need for an alternative method was necessary. Our results show that, while the Tris-NaCl-EDTA-lysozyme system (pH 8.3) worked very well with bacterial strains of Gluconobacter oxydans (ATCC9937) and Acetobacter melanogenus (NCIM2259), the Tris-sucrose-EDTA-lysozyme system worked well for Erwinia herbicola (ATCC21998), Pseudomonas chlororaphis (NCIM2041) and Corynebacterium species (ATCC31090). However, none of these systems produced spheroplasts in Brevibacterium ketosoreductum (ATCC21914), for which a separate system is under development.
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PMID:A fast spheroplast formation procedure in some 2,5-diketo-D-gluconate- and 2-keto-L-gulonate- producing bacteria. 251 94

The quantitative analysis of 2-keto-L-gulonic acid (2-KLG) produced by microbial fermentation is described. 2-KLG is separated from other aldonic and ketoaldonic acids by high-performance liquid chromatography on an Aminex anion exchange column with ammonium formate or potassium phosphate as the eluant. This is a rapid and simple method for routine analysis of a large number of samples generated by fermentation studies. Gas chromatography--mass spectrometry permits the qualitative and quantitative analysis of nanogram levels of 2-keto-L-gulonate in complex media and provides confirmation of the HPLC results. The methodologies presented are useful for the analysis of a number of aldonic and ketoaldonic acids.
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PMID:Determination of 2-keto-L-gulonic and other ketoaldonic and aldonic acids produced by ketogenic bacterial fermentation. 377 40