Nisin synthesis

Ra SR, Qiao M, Immonen T, Pujana I, Saris PEJ (1996) Genes responsible for nisin synthesis, regulation and immunity form a regulon of two operons and are induced by nisin in Lactococcus lactis N8. Microbiology 142 1281-1288... 

Nisin [1414-45-5] M 3354.2. Polypeptide from 5. lactis. Crystd from EtOH. J 52 529 1952 synthesis by Fukase et al. Tetrahedron Lett 29 795 1988.]... 

Figure 4 The biosynthesis of nisin A as a representative example of the posttranslational maturation process of lantibiotics. Following ribosomal synthesis, NisB dehydrates serine and threonine residues in the structural region of the prepeptide NisA. NisC subsequently catalyzes intramolecular addition of cysteine residues onto the dehydro amino acids in a stereo- and regioselective manner. Subsequent transport of the final product across the cell membrane by NisT and proteolytic cleavage of the leader sequence by NisP produces the mature lantibiotic. For the sequence of the leader peptide, see Figure 6. Adapted with permission from J. M. Willey W. A. van der Donk, Annu. Rev. Microbiol. 2007, 61, 477-501.

A. G. Beck-Sickinger G. Jung, Synthesis and Conformational Analysis of Lantibiotic Leader-, Pro-, and Pre-Peptides. In Nisin and Novel Lantibiotics G. Jung, H.-G. Sahl, Eds. ESCOM Leiden, 1991 pp 218-230. 

However, it was not until 1971, when Gross and Morrell reported the structure elucidation of the purified nisin peptide, 12 that it became clear that nisin contained Lan and MeLan residues. Since nisin is an important peptide antibiotic, regularly used in the food production industry as a preservative (preservative EC 234), it has attracted a great deal of attention. Nisin is produced by fermentation, however, partly because of its intriguing structure, it also became the target for the only complete synthesis of a lantibiotic reported to date in the literature. This work has been reviewed. 13 14 ... 

The synthesis of lanthionine peptides by the desulfurization of cystine peptides is exemplified by the synthesis of the nisin ring A (Scheme 2)J24 Due to the presence of two dehydroamino acids, the synthesis of ring A, together with the synthesis of rings D and E, was a considerable achievement. These syntheses were carried out in solution. A solid-phase based synthesis of methyllanthionine peptides using the desulfurization reaction has also been achieved/54 ... 

Scheme 12 Synthesis of Ring A Segment of Nisin which Contains Dehydroamino Acids and meso-Lanthio-nine1240 1...

The sulfur extrusion method 21 has been extensively investigated for the synthesis of threo-3-methyllanthionine 25 however, in this case desulfurization was not used to generate the amino acid (either in free or protected form), but rather to form different sulfide-bridged cyclic peptides as precursors for the total synthesis of nisin (3). Development of synthetic methods in this area was particularly important in respect to the synthesis of nisin, since this lantibiotic contains four // eo-3-methyllanthionine residues. The synthesis proceeded from a protected f/zreo-3-methyl-D-cysteine in the N-terminal position and L-cysteine in the C-terminal position. 

Nisin, because of its continued use as a preservative for more than 30 years 661 became an obvious target for a total synthesis 13 To date, it remains the only naturally occurring lanthionine peptide for which this feat has been achieved. In contrast to the lanthionine peptide syntheses of most other approaches already discussed, the total synthesis of nisin is based on a solution-synthesis strategy. Whilst, the total synthesis of the pentacyclic, 34-residue peptide nisin also demonstrated that such syntheses are not a commercially viable option for large amounts of lantibiotics, they have opened the way to useful synthetic methods for the synthesis of novel conformationally constrained peptides and peptidomimetic compounds. 

Scheme 19 Total Synthesis of Nisin via Condensation of Five Preformed Segments To Give Biologically Active Nisin ... 

The synthetic pentacyclic nisin was compared by analytical techniques including amino acid analysis, mass spectrometry, and NMR and was shown to be identical to natural nisin in all respects. Furthermore, the biological activity of the synthetic nisin exhibited the same activities as native nisin confirming the success of the synthesis.113,14,67-691... 

Scheme 20 Desulfurization Strategies for the Synthesis of Rings D and E (Segment IV) of Nisin...

In 1960, Ramseier [22] discovered that nisin causes leakage of intracellular molecules from cells. Later, it was shown that it disturbs the membrane potential and interferes with energy transdudion [23]. In addition, it causes inhibition of biosynthesis of the cell wall processes by blocking the synthesis of peptidoglycans [24] and by binding to the precursor lipid II [25]. However, micromolar amounts of nisin are needed to permeate artificial membranes [26,27] or to inhibit cell wall synthesis in vitro [28], while the in vivo activity of nisin is in the nanomolar range. As vancomycin, which binds to the peptide motif in lipid II, inhibited the antibacterial activity and... 

Scheme 1.1 Synthesis and structures of alkane- and alkyne-bridged nisin mimics [36].

Synthesis of Cyclic Peptide 14 an Alkyne-bridged Mimic of the Nisin A-Ring Fragment... 

DCC is also used to prereact carboxyl groups with 0-benzylhydroxylamine prior to /8-lactam ring closure. A water soluble carbodiimide in combination with HOBt is used in several steps of the total synthesis of nisin, a pentacyclic polypeptide. Carbodiimides are also used for amide bond formation in the synthesis of the antiviral antibiotic distamycin Similarly, carbodiimides are used in the synthesis of a model depsipeptide lactone related to quinoxaline antibiotics. In the last synthesis, DCC in the presence of pyridine is used in the depsipeptide bond formation. 

Insulin-dependent diabetes mellitus is an autoimmune disease and high titers of auto-antibodies against both insulin and sulfatide were found in patients with insulin-dependent diabetes (Andersson et ah, 2002 Buschard et al., 2005). Sulfatide and insulin are present in the same cellular compartments and share the same intracellular trafficking pathways (Buschard et ah, 2005 Fredman et ah, 2000). The inhibition of sulfatide synthesis with chloroquine and fumo-nisine B1 leads to inhibition of insulin granule formation in vivo (Fredman et al.,... 

Aplin Barrett). Structure contains 34 amino acid residues, eight of which are rarely found in nature, including lanthi-onine (two alanines bonded to sulfur at the 6-carbons) and P -methyllanthionine. Structure E. Gross, J. L. Morrell, J. Am. Chem. Soc. 93, 4634 (1971). Confirmation of structure of nisin and its major degradation product M. Barber et ah, Experientia 44, 266 (1988). Partial synthesis K. Fukase er ah. Bull. Chem. Soc. Japan 59, 2505 (1986). Total synthesis K, Pukase et al, Tetrahedron Letters 29, 795 (1988), Biosynthetic study G. W. Buchman et ah, J. Biol. Chem, 263, 16260 (1988). Review A. Hurst, Adv. Appl. Microbiol. 27, 85-123 (1981). 

Ingram LG (1970) A ribosomal mechanism for synthesis of peptides related to nisin. Biochim Biophys Acta224 263-265... 

Dehydroalanine (Dha) and dehydro-a-aminobutyric acid, a putative dehydration product of threonine occur accumulated in a particular class of antibiotic fungal peptides recently named lantibiotics of which Nisin (Fig. 26) will be mentioned first. The structure of the 34-peptide as revealed by Erhard Gross and coworkers in the beginning of the seventies, contains three dehydro side chains, RCH=C(NH—)CO—, and five thioether amino acids, lanthionine, Ala-S-Ala and its homolog Abu-S-Ala. Total synthesis by the group of Tetsuo Shiba (Plate 43) [61]. 

K. Fukase, M. Kitazawa, A. Sano, K. Shimbo, H. Fujita, S. Horimoto, T. Wakamiya, and T. Shiba, Total synthesis of peptide antibiotic Nisin, Tetrahedron Lett. 795-798 (1988). 

Also in Osaka, another former student of Akabori, professor Tetsuo Shiba (Plate 43) and his associates developed a procedure for the preparation of lanthionine-containing peptides and applied it, in 1988, to the synthesis of the antibiotic nisin (p. 223). In Tokyo, at Rikkyo University, professor Ichiro Muramatsu, also from the Akabori school, proposed a rapid method of synthesis and discovered novel side reactions, such as the formation of guanidine derivatives during coupling with carbodiimides. 

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