Brevibacterium ammoniagenes

Processes employing Toru/opsis yjlinus (55), Hansenu/apoljmorpha (56), Brevibacterium ammoniagenes (57), A.chromobactor butrii (58), Micrococcus lactis (59), Streptomjces testaceus (60), and others have also been patented. These procedures yield, at most, several hundred milligrams of riboflavin per Hter. 

This process has been operated successfully by the Tanabe Seiyaku Co. in Japan since 1973. Similar processes have since been commercialised by other companies, such as the Kyowa Hakko Co., often using different immobilisation methods such as polyurethane. The same iimnobilized cell approach has also been used by Tanabe since 1974 in their coimnercial process for the production of L-malic acid from fumarate using the hydratase activity of Brevibacterium ammoniagenes cells. 

Process development with Brevibacterium ammoniagenes (introduced in 1974), similarly to the development in the i-aspartate process led to 25-fold improvements, mainly through use of a Brevibacteriumflavum strain immobilized on ic-carrageenan gel with polyethyleneimine (PEI) crosslinker, so that a half life of 310 days at 37°C has been achieved. 

The first bona fide MnRNR, isolated from Corynebacterium (formerly Brevibacterium) ammoniagenes, was reported about 10 years ago [122], The MnRNR R1 protein is monomeric (dimeric in E. coli) [123]. A dinuclear Mnm—0—Mnm unit was believed to be present at its active site (cf. oxidized E. coli RNR), and its sensitivity to hydroxyurea (a radical scavenger) suggested a mechanism similar to that of its iron counterpart. However, no tyrosyl radical EPR signal was observed from the MnRNR, casting doubts about the redox role of the manganese center [118], Very recently, however, a stable EPR signal, which is inhibited by hydroxyurea and correlates directly with enzymatic activity, has been detected [124],... 

V. L. Kozel tsev, S. S. Debov and N. I. Votrin (1969). The role of inorganic polyphosphates in the formation of adenyl nucleotides in a culture of Brevibacterium ammoniagenes (in Russian). Vopr. Med. Khim., 5, 602-614. 

K. Murata, J. Kato and I. Chibata (1979). Continuous production of NADP by immobilized Brevibacterium ammoniagenes cells. Biotechnol. Bioeng., 21, 887-895. 

A similar process is also used for the production of L-malic acid from fumarate, in this case using a hydratase enzyme derived from Brevibacterium ammoniagenes. Another variation of the Tanabe technology involves the synthesis of L-alanine from L-aspartic acid through the use of immobilized whole cells (P dacunae) containing aspartate-decarboxylase. 

Fumarase. The development and use of this immobilized enzyme by Tanabe Seiyaku for production of L-malic acid is very similar to that of aspartase ( 3). Lysed Brevibacterium ammoniagenes or B. flavin cells are treated with bile acid to destroy enzymatic activity which converts fumarate to succinate. As with aspartase, the cells can be immobilized in polyacrylamide or k-carrageenan gels. Using a substrate stream of 1 M sodium fumarate at pH 7.0 and 37°C, L-malic acid of high purity has been produced since 1974 by a continuous, automated process (3,39) for example, using a 1000-L fixed-bed bioreactor, 42.2 kg L-malic acid per hour was produced continuously for 6 months. 

The third class of reductases apparently requires manganese for activity, although few other characterizations have been performed. This enzymic activity is found solely in bacteria, specifically in Brevibacterium ammoniagenes, Ar-throbacter, Micrococcus luteus, and Nocardia opaca (37, 38). 

COMPARISON OF PRODUCTIVITIES OF Brevibacterium ammoniagenes AND Brevibacterium flavum IMMOBILIZED WITH POLYACRYLAMIDE AND WITH CARRAGEENAN FOR PRODUCTION OF L-MALIC ACID... 

Other type I PanK enzymes that have been characterized are the M. tuberculosis enzyme, of which the crystal structure has been determined, and the enzyme from Corynebacterium (previously Brevibacterium) ammoniagenes ° an industrially relevant organism. 

Production of L-malic acid from fumaric acid by stereoselective hydration under the action of the intracellular fumarase in Brevibacterium ammoniagenes (Tanabe Seiyaku Co., Ltd). A 1000-liter column bioreactor can typically yield 30 tons of L-malic acid per month. 

A number of FAD analogues containing modifications in the isoalloxazine moiety have been prepared by incubation of the riboflavin analogues with the flavokinase/FAD synthetase system of Brevibacterium ammoniagenes. All... 

Brevibacterium ammoniagenes Corynebactarium equi Escherichia coli Flavobacterium capsulatum Micrococcus luteus Pseudomonas aeruginosa... 

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