Pseudomonas dacunhae immobilization

D-Aspartic acid 16 Apoxycillin Antibacterial Decarboxylation Pseudomonas dacunhae Immobilized cells [10]... 

Furui, M. and Yamashita, K. (1983) Pressurized reaction method for continuous production of L-alanine by immobilized Pseudomonas dacunhae cells. Journal of Fermentation Technology, 61, 587—591. 

I. Chibata, T. Tosa, and S. Takamatsu, Industrial production of L-alanine using immobilized Escherichia coli and Pseudomonas dacunhae, Microbiol. Sci. 1984, 1, 58-62. 

A number of decarboxylase enzymes have been described as catalysts for the preparation of chiral synthons, which are difficult to access chemically (see Chapter 2).264 The amino acid decarboxylases catalyze the pyridoxal phosphate (PLP)-dependent removal of C02 from their respective substrates. This reaction has found great industrial utility with one specific enzyme in particular, L-aspartate-P-decarboxylase (E.C. 4.1.1.12) from Pseudomonas dacunhae. This biocatalyst, most often used in immobilized whole cells, has been utilized by Tanabe to synthesize L-alanine on an industrial scale (multi-tons) since the mid-1960s (Scheme 19.33).242-265 Another use for this biocatalyst has been the resolution of racemic aspartic acid to produce L-alanine and D-aspartic acid (Scheme 19.34). The cloning of the L-aspartate-P-decarboxylase from Alcaligenes faecalis into E. coli offers additional potential to produce both of these amino acids.266... 

Alanine and aspartic acid are produced commercially utilizing enzymes. In the case of alanine, the process of decarboxylation of aspartic acid by the aspartate decarboxylase from Pseudomonas dacunhae is commercialized. The annual world production of alanine is about 200 tons. Aspartic acid is produced commercially by condensing fumarate and ammonia using aspartase from Escherichia coli. This process has been made more convenient with an enzyme immobilization technique. Aspartic acid is used primarily as a raw material with phenylalanine to produce aspartame, a noncaloric sweetener. Production and sales of aspartame have increased rapidly since its introduction in 1981. Tyrosine, valine, leucine, isoleucine, serine, threonine, arginine, glutamine, proline, histidine, cit-rulline, L-dopa, homoserine, ornithine, cysteine, tryptophan, and phenylalanine also can be produced by enzymatic methods. 

Three immobilized enzyme or microbial cell systems currently used industrially in synthesis of chiral amino acids plus one presently under development are described. L-amino acids are produced by enzymatic hydrolysis of DL-acylamino acid with aminoacylase immobilized by ionic binding to DEAE-Sephadex. Escherichia coli cells immobilized by K-carrageenan crosslinked with glutaraldehyde and hexamethylenediamine are used to convert fumaric acid and cimmonia to L-aspartic acid and Brevibacterium flavum cells similarly immobilized are used to hydrate fumaric acid to L-malic acid. The decarboxylation of L-aspcirtic acid by immobilized Pseudomonas dacunhae to L-alanine is currently under investigation. 

A continuous production system using immobilized Pseudomonas dacunhae cells with high L-aspartate 3-decarboxylase activity is currently under investigation [12]. The reaction proceeds as shown below. 

Aspartic acid, alanine, phenylalanine, and lysine were manufactured by enzymatic route. Immobilized E. coli cells expressing aspartate or the immobilized enzyme has been used in the commercial production of aspartic acid from ammonia and fumaric acid. Chibata and coworkers also produced alanine by microbial Pseudomonas dacunhae) L-aspartate P-decarboxylase with aspartate as the starting material. Phenyl alanine was manufactured from fw s-cinnamic acid and ammonia by the enzymatic route by phenyl alanine ammonia lyase as catalyst or from phenyl pyruvate and aspartic acid using transaminase. 

Senuma, M., Otsuki, O., Sakata, N., Furui, M., Tosa, T, 1989. Industrial production of D-aspartic acid and L-alanine from DL-aspartic acid using a pressurized column reactor containing immobilized Pseudomonas dacunhae cells. Journal of Fermentation and Bioengineering 67,233. 

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