Enzymatic Synthesis of Streptidine from scyllo-Inosamine<sup>*</sup>

Walker, James B.; Walker, Margaret S.

doi:10.1021/bi00864a028

Cited by 47 publications

(37 citation statements)

References 14 publications

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“…Aminodeoxy-scyllo-inositol 4-kinase activity was detected in extracts of mature mycelia of S. glebosus (Fig. 3), just as in the case of streptomycin producers (29). Reaction (Table 1).…”

Section: Resultsmentioning

confidence: 72%

“…1, I) have provided evidence that an unusual sequence of five reactions converts an inositol hydroxyl group to the first guanidino group; the second guanidino group is then formed by a second mechanistically similar sequence of five reactions (21,28,29,31,(33)(34)(35). Studies of mutants blocked in streptidine biosynthesis have subsequently provided independent support for the proposed enzymatic pathway (6,12,15).…”

mentioning

confidence: 99%

“…Unless otherwise noted, most assays reported consisted of 5 ,ul of radioactive substrate (15, (29,34).…”

mentioning

confidence: 99%

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Possible evolutionary relationships between streptomycin and bluensomycin biosynthetic pathways: detection of novel inositol kinase and O-carbamoyltransferase activities

Walker

1990

J Bacteriol

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Bluensomycin (glebomycin) is an aminocyclitol antibiotic that differs structurally from dihydrostreptomycin in having bluensidine (lD-1-O-carbamoyl-3-guanidinodeoxy-scyllo-inositol) rather than streptidine (1,3-diguanidino-1,3-dideoxy-scyllo-inositol) as its aminocyclitol moiety. Extracts of the bluensomycin producer Streptomyces hygroscopicus form glebosus ATCC 14607 (S. glebosus) were found to have aminodeoxy-scylloinositol kinase activity but to lack 1D-1-guanidino-3-amino-1,3-dideoxy-scyllo-inositol kinase activity, showing for the first time that these two reactions in streptomycin producers must be catalyzed by different enzymes.S. glebosus extracts therefore possess the same five enzymes required for synthesis of guanidinodeoxy-scylloinositol from myo-inositol that are found in streptomycin producers but lack the next three of the four enzymes found in streptomycin producers that are required to synthesize the second guanidino group of streptidine-P.In place of a second guanidino group, S. glebosus extracts were found to catalyze a Mg2e-dependent carbamoylation of guanidinodeoxy-scyllo-inositol to form bluensidine, followed by a phosphorylation to form bluensidine-P. The novel carbamoyl-P:guanidinodeoxy-scyllo-inositol O-carbamoyltransferase and ATP:bluensidine phosphotransferase activities were not detected in streptomycin producers or in S. glebosus during its early rapid growth phase. Free bluensidine appears to be a normal intermediate in bluensomycin biosynthesis, in contrast to the case of streptomycin biosynthesis; in the latter, although exogenous streptidine can enter the pathway via streptidine-P, free streptidine is not an intermediate in the endogenous biosynthetic pathway. Comparison of the streptomycin and bluensomycin biosynthetic pathways provides a unique opportunity to evaluate those proposed mechanisms for the evolutionary acquisition of new biosynthetic capabilities that involve gene duplication and subsequent mutational changes in one member of the pair. In this model, there are at least five pairs of enzymes catalyzing analogous reactions that can be analyzed for homology at both the protein and DNA levels, including two putative pairs of inositol kinases detected in this study.Actinomycete soil bacteria have a large excess of DNA in their genomes over that required for growth and morphological differentiation. Much of this excess DNA codes for enzymes responsible for the unique biosynthetic abilities of actinomycetes, which as a group produce 70% of the known antibiotics plus numerous other idiolites (secondary metabolites). The mechanisms and selection procedures by which the diverse pathways for idiolite biosynthesis have evolved and are maintained are problems of both theoretical and practical interest. Model systems for addressing these problems must be amenable to both enzymatic and genetic analysis. The streptomycin biosynthetic pathway fulfills these two requirements for a model system (32).Enzymatic studies on biosynthesis of the streptidine moiety of dihydrostreptomycin (Fi...

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“…Aminodeoxy-scyllo-inositol 4-kinase activity was detected in extracts of mature mycelia of S. glebosus (Fig. 3), just as in the case of streptomycin producers (29). Reaction (Table 1).…”

Section: Resultsmentioning

confidence: 72%

mentioning

confidence: 99%

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Possible evolutionary relationships between streptomycin and bluensomycin biosynthetic pathways: detection of novel inositol kinase and O-carbamoyltransferase activities

Walker

1990

J Bacteriol

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“…of the streptidine moiety of streptomycin and hydroxystreptomycin and the bluensidine moiety of bluensomycin (glebomycin), previously discovered by our laboratory (16,(18)(19)(20)24 -amino groups of certain diaminocyclitols by the reactions of equations 9 to 12. The reactions are generalized here for streptamine synthesis as might occur in the synthesis of the SS-56C antibiotic (7) or 2-hydroxygentamicin by Daum's idiotroph (5,6).…”

Section: Fig 2 Enzymatic Reactions Involved In Biosynthesis From Mymentioning

confidence: 98%

“…1). The positions were established by our earlier work with streptamine and 2-deoxystreptamine (20,23,24). The results are shown in Table 3.…”

Section: Myo-[mentioning

confidence: 99%

Enzymatic synthesis of aminocyclitol moieties of aminoglycoside antibiotics from inositol by Streptomyces spp.: detection of glutamine-aminocyclitol aminotransferase and diaminocyclitol aminotransferase activities in a spectinomycin producer

Walker

1995

J Bacteriol

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Structural, Functional and Calorimetric Investigation of MosA, a Dihydrodipicolinate Synthase from Sinorhizobium meliloti L5–30, does not Support Involvement in Rhizopine Biosynthesis

et al. 2008

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MosA is an enzyme from Sinorhizobium meliloti L5-30, a beneficial soil bacterium that forms a symbiotic relationship with leguminous plants. MosA was proposed to catalyze the conversion of scyllo-inosamine to 3-O-methyl-scyllo-inosamine (compounds known as rhizopines), despite the MosA sequence showing a strong resemblance to dihydrodipicolinate synthase (DHDPS) sequences rather than to methyltransferases. Our laboratory has already shown that MosA is an efficient catalyst of the DHDPS reaction. Here we report the structure of MosA, solved to 1.95 A resolution, which resembles previously reported DHDPS structures. In this structure Lys161 forms a Schiff base adduct with pyruvate, consistent with the DHDPS mechanism. We have synthesized both known rhizopines and investigated their ability to interact with MosA in the presence and absence of methyl donors. No MosA-catalyzed methyltransferase activity is observed in the presence of scyllo-inosamine and S-adenosylmethionine (SAM). 2-Oxobutyrate can form a Schiff base with MosA, acting as a competitive inhibitor of MosA-catalyzed dihydrodipicolinate synthesis. It can be trapped on the enzyme by reaction with sodium borohydride, but does not act as a methyl donor. The presence of rhizopines does not affect the kinetics of dihydrodipicolinate synthesis. Isothermal titration calorimetry (ITC) shows no apparent interaction of MosA with rhizopines and SAM. Similar experiments with pyruvate as titrant demonstrate that the reversible Schiff base formation is largely entropically driven. This is the first use of ITC to study Schiff base formation between an enzyme and its substrate.

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Enzymatic Synthesis of Streptidine from scyllo-Inosamine^*

Cited by 47 publications

References 14 publications

Possible evolutionary relationships between streptomycin and bluensomycin biosynthetic pathways: detection of novel inositol kinase and O-carbamoyltransferase activities

Possible evolutionary relationships between streptomycin and bluensomycin biosynthetic pathways: detection of novel inositol kinase and O-carbamoyltransferase activities

Enzymatic synthesis of aminocyclitol moieties of aminoglycoside antibiotics from inositol by Streptomyces spp.: detection of glutamine-aminocyclitol aminotransferase and diaminocyclitol aminotransferase activities in a spectinomycin producer

Structural, Functional and Calorimetric Investigation of MosA, a Dihydrodipicolinate Synthase from Sinorhizobium meliloti L5–30, does not Support Involvement in Rhizopine Biosynthesis

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Enzymatic Synthesis of Streptidine from scyllo-Inosamine*

Cited by 47 publications

References 14 publications

Possible evolutionary relationships between streptomycin and bluensomycin biosynthetic pathways: detection of novel inositol kinase and O-carbamoyltransferase activities

Possible evolutionary relationships between streptomycin and bluensomycin biosynthetic pathways: detection of novel inositol kinase and O-carbamoyltransferase activities

Enzymatic synthesis of aminocyclitol moieties of aminoglycoside antibiotics from inositol by Streptomyces spp.: detection of glutamine-aminocyclitol aminotransferase and diaminocyclitol aminotransferase activities in a spectinomycin producer

Structural, Functional and Calorimetric Investigation of MosA, a Dihydrodipicolinate Synthase from Sinorhizobium meliloti L5–30, does not Support Involvement in Rhizopine Biosynthesis

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Enzymatic Synthesis of Streptidine from scyllo-Inosamine^*