Actinoplanes utahensis

ECB can be cleaved by a deacylating enzyme ( ECB deacylase ) from a Pseudomonas sp. [23] or from Actinoplanes utahensis [24], generating ECB nu-... 

ECB deacylase is produced naturally by Actinoplanes utahensis. However, very little (—0.2%) deacylase activity was detected in the culture filtrate of Actinoplanes utahensis. Less than 5% of the cell-associated deacylase activity was released by incubation of the cells at 0.01 M KH2P04, pH 6, for 1 day. A simple salt treatment with 0.8 M KC1 resulted in 60-80% recovery of soluble deacylase at a high specific activity [25], This salt-induced solubilization suggests that the deacylase is loosely bound to the membrane of A. utahensis and can be released by disruption of ionic interactions [26]. The solubilized enzyme was stable and purified to apparent homogeneity by a four-step conventional procedure [25]. 

LD Boeck, DS Fukuda, BJ Abbott, M Debono. Deacylation of A21978C, an acidic lipopeptide antiobiotic complex, by Actinoplanes utahensis. J Antibiot 41 1085-1092, 1988. 

AJ Kreuzman, RL Hodges, JR Swartling, TE Pohl, SK Ghag, PJ Baker, D McGil-vray, WK Yeh. Membrane-associated echinocandin B deacylase of Actinoplanes utahensis. purification, characterization, heterologous cloning and enzymatic deacylation reaction. J Ind Microbiol Biotechnol 24 173-180, 2000. 

J Inokoshi, H Takeshima, H Ikeda, S Omura. Cloning and sequencing of the aculea-cin A acylase-encoding gene from Actinoplanes utahensis and expression in Streptomyces lividans. Gene 119 29-35, 1992. 

H Takeshima, J Inokoshi, Y Takada, H Tanaka, S Omura. A deacylation enzyme for aculeacin A, a neutral lipopeptide antibiotic, from Actinoplanes utahensis purification and characterization. J Biochem 105 606-610, 1989. 

A21978C core peptide. Lilly scientists reasoned that, if they could remove the lipid side chain, it would open up the molecule to facile SAR studies with other lipid side chains. They discovered that Actinoplanes utahensis produced a deacylase enzyme that cleaved the lipid side chains from all A21978C factors.10 This provided a robust route to prepare the core peptide for chemical modification. 

Initial modification of the acyl side chain yielded FR131535.11 The synthesis of this novel echinocandin-like lipopeptide is outlined in Figure 15.5. The palmitoyl group was removed from FR901379 by treating it with acylase from Actinoplanes utahensis, which yielded FR179642. A new acyl side chain was prepared starting with 1-bromooctane and 4-hydroxybenzoic acid. 

S. (S.A.) CjgHjoO.j, Mr 574.54, mp. 257-265 C. [a] +85 (CH3OH), orange-yellow crystals. An an-thracycline antibiotic from the actinomycetes Strepto-myces steffisburgensis, S. elgreteus, and Actinoplanes utahensis exhibiting antibacterial and carcinostatic activities. 

C28H30O13 574.537 Anthracycline antibiotic, Isol. from Streptomyces steffisburgensis, Strepto-myces elgreteus and Actinoplanes utahensis. Antibacterial and antineo-plastic agent. Orange-yellow cryst. 

Analogous lipopeptides were prepared from A21978 C by enzymatic hydrolysis using Actinoplanes utahensis and reacylation after suitable protection of the peptide, for evaluation of their antibiotic properties (727). The n-decanoyl analogue had the lowest level of toxicity and was called daptomycin. 

Peptidolipid A21978 C displayed antibiotic activity against Gram positive bacteria. Enzymatic hydrolysis by incubation with Actinoplanes utahensis resulting in removal of the fatty acid was followed by chemical attachment of another fatty acid. Of the various analogues prepared in this manner, the n-decanoyl analogue (daptomycin) had the least acute toxicity in the mouse and exhibited antimicrobial properties at about 0,5 ig/ml (127). 

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