Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound having a 1-thia-5-aza-bicyclo
Patent
1996-05-13
1998-08-18
Hendricks, Keith D.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing compound having a 1-thia-5-aza-bicyclo
435 46, 435 47, 4352545, 4352523, 536 232, 536 241, C12P 3502, C12P 3702, C12N 115
Patent
active
057957339
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION AND BRIEF DESCRIPTION OF THE PRIOR ART
The present invention concerns a biosynthetic process for preparation and recovery of 7-aminodesacetoxycephalosporanic acid (7-ADCA).
.beta.-Lactam antibiotics constitute the most important group of antibiotic compounds, with a long history of clinical use. Among this group, the prominent ones are the penicillins and cephalosporins. These compounds are naturally produced by the filamentous fungi Penicillium chrysogenum and Acremonium chrysogenum, respectively.
As a result of classical strain improvement techniques, the production levels of the antibiotics in Penicillium chrysogenum and Acremonium chrysogenum have increased dramatically over the past decades. With the increasing knowledge of the biosynthetic pathways leading to penicillins and cephalosporins, and the advent of recombinant DNA technology, new tools for the improvement of production strains and for the in vivo derivatization of the compounds have become available.
Most enzymes involved in .beta.-lactam biosynthesis have been identified and their corresponding genes been cloned, as can be found in Ingolia and Queener, Med. Res. Rev. 9 (1989), 245-264 (biosynthesis route and enzymes), and Aharonowitz, Cohen, and Martin, Ann. Rev. Microbiol. 46 (1992), 461-495 (gene cloning).
The first two steps in the biosynthesis of penicillin in P. chrysogenum are the condensation of the three amino acids L-5-amino-5-carboxypentanoic acid (L-.alpha.-aminoadipic acid) (A), L-cysteine (C) and L-valine (V) into the tripeptide LLD-ACV, followed by cyclization of this tripeptide to form isopenicillin N. This compound contains the typical .beta.-lactam structure.
The third step involves the exchange of the hydrophilic side chain of L-5-amino-5-carboxypentanoic acid by a hydrophobic side chain by the action of the enzyme acyltransferase (AT). In the industrial process for penicillin G production the side chain of choice is phenylacetic acid (PA). In EP-A-0532341 the application of an adipate (5-carboxypentanoate) feedstock has been disclosed. The incorporation of this substrate leads to a penicillin derivative with a 5-carboxypentanoyl side chain, viz. 5-carboxypentanoyl-6-aminopenicillanic acid. This incorporation is due to the fact that the acyltransferase has a proven wide substrate specificity (Behrens et al., J. Biol. Chem. 175 (1948), 751-809; Cole, Process. Biochem. 1 (1966), 334-338; Ballio et al., Nature 185 (1960), 97-99). The enzymatic exchange reaction mediated by AT takes place inside a cellular organelle, the microbody, as has been described in EP-A-0448180.
Cephalosporins are much more expensive than penicillins. One reason is that some cephalosporins (e.g. cephalexin) are made from penicillins by a number of chemical conversions. Another reason is that, so far, only cephalosporins with a D-5-amino-5-carboxypentanoyl side chain can be fermented. Cephalosporin C, by far the most important starting material in this respect, is very soluble in water at any pH, thus implying lengthy and costly isolation processes using cumbersome and expensive column technology. Cephalosporin C obtained in this way has to be converted into therapeutically used cephalosporins by a number of chemical and enzymatic conversions.
The intermediate 7-ADCA is currently produced by chemical derivatization of penicillin G. The necessary chemical steps to produce 7-ADCA involve the expansion of the 5-membered penicillin ring structure to a 6-membered cephalosporin ring structure. However, the expandase enzyme from the filamentous bacterium Streptomyces clavuligerus can carry out such ring expansions. When introduced into P. chrysogenum, it can convert the penicillin ring structure into the cephalosporin ring structure, as described in Cantwell et al. , Proc. R. Soc. Lond. B. 248 (1992), 283-289; and in EP-A-0532341 and EP-A-0540210. The expandase enzyme has been well characterized (EP-A-0366354) both biochemically and functionally, as has its corresponding gene. Both physical maps of the cefE gene (EP-A-0233715), DNA seq
REFERENCES:
patent: 5070020 (1991-12-01), Ingolla et al.
Baldwin, J. et al., Tetrahedron, vol. 43, No. 13, pp. 3009-3014 (1987).
Ballio, A. et al., Nature, vol. 185, pp. 97-99 (1960).
Coque, J.J. R. et al., Molecular and General Genetics, vol. 236, pp. 453-458 (1993).
Muller et al (1992) Bioc Biop Acta 1116:210-213. "Involvement of Microbodies in Penicillin Biosynthesis".
Maeda et al (1995) Enz. Microb. Tech. 17:231-234 The Substrate Specificity of Deacetoxycephalosporin C Synthase ("Expanse").
Bowers et al (1984) Bioc. Biophys. Res. Comm. 120:607-613 "Enzymatic Synthesis of the Penicilln and Cephalosporin . . . ".
Bovenberg Roelof Ary Lans
De Vroom Erik
Hoekema Andreas
Koekman Bertus Pieter
Van Der Laan Jan Metske
Gist-Brocades B.V.
Hendricks Keith D.
LandOfFree
Process for the efficient production of 7-ADCA via 3-(carboxyeth does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Process for the efficient production of 7-ADCA via 3-(carboxyeth, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Process for the efficient production of 7-ADCA via 3-(carboxyeth will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-1113744