Polypeptide Antibiotic Research Articles

Hypopulvins, novel peptaibiotics from the polyporicolous fungus Hypocrea pulvinata, are produced during infection of its natural hosts

In order to investigate the significance of antibiotics for the producing organism(s) in the natural habitat, we screened specimens of the polyporicolous fungus Hypocrea pulvinata growing on its natural hosts Piptoporus betulinus and Fomitopsis pinicola. Results showed that a particular group of nonribosomally biosynthesised antibiotic polypeptides, the peptaibiotics, which contain the nonproteinogenic marker amino acid α-aminoisobutyric acid (Aib), was produced in the natural habitat by the fungicolous producer and, consequently, released into the host. Using liquid chromatography coupled to electrospray high-resolution mass spectrometry we detected especially 19-, but also 11-, 18-, and 20-residue peptaibiotics in the five infected specimens analysed. Structures of peptaibiotics found were confirmed by analysing the peptaibiome of pure agar cultures obtained by single-ascospore isolation from the specimens. The 19-residue peptaibols were determined as deletion sequences of the trichosporins B lacking the Aib residue in position 6. Notably, 26 of the 28 peptaibiotics sequenced were novel; therefore the name ‘hypopulvins’ was introduced. Considering not only the ubiquity of both the two host species but also the highly specific association between H. pulvinata and P. betulinus/F. pinicola, and the abundance of this fungicolous species in north temperate regions of the world, a decisive role for the peptaibiotics detected in this study is predicted, which may act as mediators of the complex interactions between the basidiomycetous host and its fungicolous ascomycete ‘partner’. Structural analogies of the hypopulvins, particularly with other 18-, 19-, and 20-residue peptaibiotics, suggest that the hypopulvins are forming transmembrane ion channels and could thus support the hypothesis of a parasitic lifestyle of the fungicolous producer.

Role of Heme Oxygenase-1 in Polymyxin B-Induced Nephrotoxicity in Rats

Polymyxin B (PMB) is a cationic polypeptide antibiotic with activity against multidrug-resistant Gram-negative bacteria. PMB-induced nephrotoxicity consists of direct toxicity to the renal tubules and the release of reactive oxygen species (ROS) with oxidative damage. This study evaluated the nephroprotective effect of heme oxygenase-1 (HO-1) against PMB-induced nephrotoxicity in rats. Adult male Wistar rats, weighing 286 ± 12 g, were treated intraperitoneally once a day for 5 days with saline, hemin (HO-1 inducer; 10 mg/kg), zinc protoporphyrin (ZnPP) (HO-1 inhibitor; 50 μmol/kg, administered before PMB on day 5), PMB (4 mg/kg), PMB plus hemin, and PMB plus ZnPP. Renal function (creatinine clearance, Jaffe method), urinary peroxides (ferrous oxidation of xylenol orange version 2 [FOX-2]), urinary thiobarbituric acid-reactive substances (TBARS), renal tissue thiols, catalase activity, and renal tissue histology were analyzed. The results showed that PMB reduced creatinine clearance (P < 0.05), with an increase in urinary peroxides and TBARS. The PMB toxicity caused a reduction in catalase activity and thiols (P < 0.05). Hemin attenuated PMB nephrotoxicity by increasing the catalase antioxidant activity (P < 0.05). The combination of PMB and ZnPP incremented the fractional interstitial area of renal tissue (P < 0.05), and acute tubular necrosis in the cortex area was also observed. This is the first study demonstrating the protective effect of HO-1 against PMB-induced nephrotoxicity.

Mechanism-independent method for predicting response to multidrug combinations in bacteria

Drugs are commonly used in combinations larger than two for treating bacterial infection. However, it is generally impossible to infer directly from the effects of individual drugs the net effect of a multidrug combination. Here we develop a mechanism-independent method for predicting the microbial growth response to combinations of more than two drugs. Performing experiments in both Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus) bacteria, we demonstrate that for a wide range of drugs, the bacterial responses to drug pairs are sufficient to infer the effects of larger drug combinations. To experimentally establish the broad applicability of the method, we use drug combinations comprising protein synthesis inhibitors (macrolides, aminoglycosides, tetracyclines, lincosamides, and chloramphenicol), DNA synthesis inhibitors (fluoroquinolones and quinolones), folic acid synthesis inhibitors (sulfonamides and diaminopyrimidines), cell wall synthesis inhibitors, polypeptide antibiotics, preservatives, and analgesics. Moreover, we show that the microbial responses to these drug combinations can be predicted using a simple formula that should be widely applicable in pharmacology. These findings offer a powerful, readily accessible method for the rational design of candidate therapies using combinations of more than two drugs. In addition, the accurate predictions of this framework raise the question of whether the multidrug response in bacteria obeys statistical, rather than chemical, laws for combinations larger than two.

Polymyxin Use Associated With Respiratory Arrest

The polymyxins (polymyxin B and E) are bactericidal polypeptide antibiotics first discovered in 1947 and used for the treatment of gram-negative bacterial infections. Renal and neurologic toxicities coupled with the increasing availability of effective alternatives led to declining use in the 1960s. The emergence of multidrug-resistant organisms in the past decade has resulted in a resurgence in the use of polymyxins in critically ill patients, yet the side effects are not well known. We report two cases of respiratory arrest likely due to polymyxin B infusions in the context of a 10-fold increase in the use of polymyxin B in our institution over the past 10 years.

Neonatal Ventriculitis with Multi Drug Resistant Acinetobactor baumanii: A Case Report and Review of Literature

Meningitis due to Multi Drug Resistant Acinetobacter baumannii (MRAB) is a serious emerging problem in pediatric intensive care. Treatment of these patients is challenging and polypeptide antibiotic is an important option but have poor cerebrospinal fluid (CSF) penetration. We describe our experience of a successfully treated neonatal ventriculitis and meningitis due to MRAB with intraventricular polymyxin B.

Targeted Killing of Escherichia Coli by an Unfolded Protein

The appearance of antibiotic resistant and novel pathogenic strains of E. coli emphasises the continuing need for new antibacterial treatments. Many small proteins exist in nature that kill bacteria by disrupting their membranes but, since human and bacterial lipid bilayers are similar, it is difficult to avoid toxicity to the patient. We study Colicin N (ColN), a ∼ 400 residue globular protein which contains a short unfolded N-terminal domain. The folded region binds to bacteria and kills by forming pores in the inner bacterial membrane. The unfolded N-terminal 90 amino acids (ColN-T) were thought simply to help translocate the toxic domain to the inner-membrane. Here we report the unexpected finding that, in isolation, ColN-T is bactericidal and, although less efficient than ColN, is strictly dependent upon the same receptor proteins, OmpF and TolA. The binding sites for these occupy either end of the linear polypeptide. Also like ColN, ColN-T induces K+ leakage across the inner-membrane. However, unlike most antimicrobial peptides, ColN-T does not disrupt, nor bind to, model lipid bilayers. These data imply that, uniquely, it can induce E. coli killing by OmpF/TolA interactions alone. Since these receptors are only found in E. coli-like bacteria, ColN-T displays the unusual combination of a generic killing mechanism coupled with extreme specificity. As a result, we believe that this unstructured polypeptide antibiotic could point the way to wholly new targets in antibacterial therapy.

Renal and neurological side effects of colistin in critically ill patients

Colistin is a complex polypeptide antibiotic composed mainly of colistin A and B. It was abandoned from clinical use in the 1970s because of significant renal and, to a lesser extent, neurological toxicity. Actually, colistin is increasingly put forward as salvage or even first-line treatment for severe multidrug-resistant, Gram-negative bacterial infections, particularly in the intensive care setting. We reviewed the most recent literature on colistin treatment, focusing on efficacy and toxicity issues. The method used for literature search was based on a PubMed retrieval using very precise criteria.Despite large variations in dose and duration, colistin treatment produces relatively high clinical cure rates. Colistin is potentially nephrotoxic but currently used criteria tend to overestimate the incidence of kidney injury. Nephrotoxicity independently predicts fewer cures of infection and increased mortality. Total cumulative colistin dose is associated with kidney damage, suggesting that shortening of treatment duration could decrease the incidence of nephrotoxicity. Factors that may enhance colistin nephrotoxicity (i.e., shock, hypoalbuminemia, concomitant use of potentially nephrotoxic drugs) must be combated or controlled. Neurotoxicity does not seem to be a major issue during colistin treatment. A better knowledge of colistin pharmacokinetics in critically ill patients is imperative for obtaining colistin dosing regimens that ensure maximal antibacterial activity at minimal toxicity.

The Preparation and Resolution of Acridyl(5)—N. Alanine Ethyl Ester

Abstract It has been reported that D amino-acids are present in the molecules of various antibiotic polypeptides such as gramicidin1,2, tyrociain3, gramicidin S4, aerosporin5 and bacitracin6. The penicillins on degradation give D-penicillamine (β:β-dimethylcysteine) and du Vigneaud and his collaborators7 have shown that an antibiotic penicillin may be synthesised from D-penicillamine, whereas the isomer from L-penicillamine is biologically inactive.

Cutting Edge: Cyclic Polypeptide and Aminoglycoside Antibiotics Trigger IL-1β Secretion by Activating the NLRP3 Inflammasome

Clinical use of antibiotics is based on their capacity to inhibit bacterial growth via bacteriostatic or bacteriocidal effects. In this article, we show that the aminoglycoside antibiotic neomycin, the cyclic lipopeptide antibiotic polymyxin B, and the cyclic peptide antibiotics gramicidin and tyrothricin can induce IL-1β secretion in bone marrow dendritic cells and macrophages. LPS priming was required to trigger the transcription and translation of pro-IL-1β but was independent of TNFR or IL-1R signaling. All four antibiotics required the NLRP3 inflammasome, the adaptor ASC, and caspase-1 activation to secrete IL-1β, a process that depended on potassium efflux but was independent of P2X7 receptor. All four antibiotics induced neutrophil influx into the peritoneal cavity of mice, which required NLRP3 only in the case of polymyxin B. Together, certain antibiotics have the potential to directly activate innate immunity of the host.

The BceABRS Four-Component System Regulates the Bacitracin-Induced Cell Envelope Stress Response in Streptococcus mutans

Streptococcus mutans is known to be resistant to bacitracin, a cyclic polypeptide antibiotic produced by certain species of the genus Bacillus. This property is often exploited in the isolation of S. mutans strains from highly heterogeneous oral microflora. A genetic locus consisting of a four-gene operon, bceABRS (formerly mbrABCD), the component genes of which are homologous to Bacillus subtilis bceRS-bceAB (encoding a two-component system and an ABC transporter), is required for bacitracin resistance in S. mutans. Here we describe the identification of a DNA binding site for the BceR response regulator and its transcriptional control of the bceABRS operon in response to the presence of bacitracin. We provide evidence indicating that phosphorylated BceR binds directly to a conserved invert repeat located between bp -120 and -78 of the bceABRS promoter region and positively regulates expression of the bceABRS operon. We also demonstrate that sensing of bacitracin by the BceS histidine kinase requires the presence of an intact BceAB transporter, since deletion of either bceA or bceB abolishes BceRS-mediated bacitracin sensing. The results suggest that the BceAB transporter acts as a cosensor, together with the BceRS two-component system, for bacitracin perception in S. mutans. By searching the S. mutans genome databases, we have identified three additional genes that share the consensus BceR binding motif at their promoter regions. Our initial work has confirmed that expression of these genes is directly controlled by BceRS, indicating that the bceABRS operon, along with the three additional genes, forms the BceRS regulon in S. mutans. Taking these findings together, we conclude that BceABRS comprises a four-component system that plays an important role in stimulus sensing, signal transduction, the gene regulatory network, and substrate transport for the cell envelope stress response in S. mutans.

2D IR Line Shapes Probe Ovispirin Peptide Conformation and Depth in Lipid Bilayers

We report a structural study on the membrane binding of ovispirin using 2D IR line shape analysis, isotope labeling, and molecular dynamics simulations. Ovispirin is an antibiotic polypeptide that binds to the surfaces of membranes as an alpha-helix. By resolving individual backbone vibrational modes (amide I) using 1-(13)C=(18)O labeling, we measured the 2D IR line shapes for 15 of the 18 residues in this peptide. A comparison of the line shapes reveals an oscillation in the inhomogeneous line width that has a period equal to that of an alpha-helix (3.6 amino acids). The periodic trend is caused by the asymmetric environment of the membrane bilayer that exposes one face of the alpha-helix to much stronger environmental electrostatic forces than the other. We compare our experimental results to 2D IR line shapes calculated using the lowest free energy structure identified from molecular dynamics simulations. These simulations predict a periodic trend similar to the experiment and lead us to conclude that ovispirin lies in the membrane just below the headgroups, is tilted, and may be kinked. Besides providing insight into the antibiotic mechanism of ovispirin, our procedure provides an infrared method for studying peptide and protein structures that relies on the natural vibrational modes of the backbone. It is a complementary method to other techniques that utilize line shapes, such as fluorescence, NMR, and ESR spectroscopies, because it does not require mutations, the spectra can be quantitatively simulated using molecular dynamics, and the technique can be applied to difficult-to-study systems like ion channels, aggregated proteins, and kinetically evolving systems.

An approach to on-line electrospray mass spectrometric detection of polypeptide antibiotics of enramycin for high-speed counter-current chromatographic separation

In the field of pharmaceutical and biomedical analysis of peptides, a rapid on-line detection and identification for a methodology have been required for the discovery of new biological active products. In this study, a high-speed counter-current chromatography with electrospray mass spectrometry (HSCCC/ESI-MS) was developed for the on-line detection and purification of polypeptide antibiotics of enramycin-A and -B. The analytes were purified on HSCCC model CCC-1000 (multi-layer coil planet centrifuge) with a volatile solvent of two-phase system composed of n-butanol/hexane/0.05% aqueous trifluoroacetic acid solution (43/7/50, V/V/V), and detected on an LCMS-2010EV quadrupole mass spectrometer fitted with an ESI source system in positive ionization following scan mode ( m/ z 100–2000). The HSCCC/ESI-MS peaks indicated that enramycin-A (major m/ z 786 [M+3H] 3+ and minor m/ z 1179 [M+2H] 2+) and enramycin-B (major m/ z 791 [M+3H] 3+ and minor m/ z 1185 [M+2H] 2+) have the peak resolution value of 2.9 from 15 mg of loaded enramycin powder. The HSCCC collected amounts of the peak fractions were additionally 4.3 mg (enramycin-A), and 5.9 mg (enramycin-B), respectively. These purified substances were analyzed by LC/ESI-MS with scan positive mode. Based on the LC/ESI-MS chromatograms and spectra of the fractions, enramycin-A and -B were estimated to be over 95% purity. The overall results indicate that this approach of HSCCC/ESI-MS is a powerful technique for the purification and identification of bioactive peptides.

Occurrence and characterization of peptaibols from Trichoderma citrinoviride, an endophytic fungus of cork oak, using electrospray ionization quadrupole time-of-flight mass spectrometry

A cork oak endophytic strain of Trichoderma citrinoviride, previously selected for its antagonistic potential against various fungal pathogens involved in oak decline, was screened for the production of bioactive secondary metabolites. From liquid culture a mixture of polypeptide antibiotics (peptaibols) belonging to the paracelsin family was isolated and characterized. This peptide mixture was purified by column chromatography and preparative TLC on silica gel, and separated by analytical HPLC. It was analysed by MALDI-TOF MS and nano-ESI-QTOF MS. Tandem mass experiments were performed to determine the amino acid sequences based on the fragmentation pattern of selected parent ions. The mixture comprised 20-residue peptides with C-terminal phenylalaninol and N-terminal acetylation. Twenty-eight amino acid sequences were identified, and amino acid exchanges were located in positions 6, 9, 12 and 17. Among them, seven sequences are new as compared to those reported in the database specifically for peptaibols and in the literature. In addition, we obtained experimental evidence suggesting the existence of non-covalent dimeric forms (homo- and hetero-) of the various peptaibol species. The peptide mixture showed strong antifungal activity toward seven important forest tree pathogens, and it was highly toxic in an Artemia salina (brine shrimp) bioassay. These results emphasize the cryptic role of endophytic fungi as a source of novel bioactive natural products and biocontrol agents.

Use of Colistin in Children

Colistin, or Polymixin E, is a cationic polypeptide antibiotic obtained from Bacillus spp. It was initially used therapeutically in the United States in 1959 but intravenous formulations were gradually abandoned in the early 1980s due to nephrotoxicity concerns. The emergence of bacteria resistant to most classes of commercially available antibiotics and the shortage of new antimicrobial agents with activity against gram-negative microorganisms have led to the reconsideration of polymixins as valuable therapeutic options.

Colistin and polymyxin B: A re-emergence

One of the greatest achievements of modern medicine is the development of antibiotics against life-threatening infections, but the emergence of multidrug-resistant (MDR) gram negative bacteria has drastically narrowed down the therapeutic options against them. This limitation has led clinicians to reappraise the clinical application of polymyxins, an old class of cationic, cyclic polypeptide antibiotics. Polymyxins are active against selected gram-negative bacteria, including the Acinetobacter species, Pseudomonas aeruginosa, Klebsiella species, and Enterobacter species. In this article, we summarise the chemistry, pharmacokinetics, and pharmacodynamics of polymyxins and the latest understanding of their action against MDR pathogens.

Studies on the Selective Trifluoroacetolytic Scission of Native Peptaibols and Model Peptides Using HPLC and ESI‐CID‐MS

Representative members of a group of linear, N-acylated polypeptide antibiotics (peptaibols) containing alpha-aminoisobutyric acid (Aib) and, in part, isovaline (Iva), as well as proteinogenic amino acids and a C-terminal-bonded 2-amino alcohol, were treated with anhydrous trifluoroacetic acid (TFA) at 37 degrees for 0.5-26 h. The resulting fragments were separated by HPLC and characterized by electrospray ionization collision-induced dissociation mass spectrometry (ESI-CID-MS). The following 16-20-residue peptaibols were investigated: natural, microheterogeneous mixtures of antiamoebins and alamethicin F50, uniform paracelsin A, and synthetic trichotoxin A50/E. In the natural peptides, bonds formed between Aib (Iva) and Pro (Hyp) were rapidly and selectively cleaved within 0.5 h. Furthermore, TFA esters of the C-terminal amino alcohols were formed. Depending on time, release of C-terminal tri- and tetrapeptides as well as amino acids from the major fragments was observed. Synthetic homooligopeptides, namely Z- and Ac-(Aib)(10)-O(t)Bu and Z-(Aib)(7)-O(t)Bu, were analyzed for comparison. On treatment with TFA, a regular series of Z-(Aib)(10-5)-OH from Z-(Aib)(10)-O(t)Bu were detected within 0.5 h, and, after 3 h, release of a regular series of Z-(Aib)(7-3)-OH from Z-(Aib)(7)-O(t)Bu were observed. Moreover, concomitant release of the series of H-(Aib)(10-3)-OH from the decapeptide occurred. From these data, a repetitive cleavage mechanism via intermediate formation of C-terminal oxazolones on trifluoroacetolysis is proposed. Furthermore, their formation and stability in native peptaibols are correlated with subtle structural differences in protein amino acids linked to Aib. From the conspicuous concordance of the formation and abundance of regular series of trifluoroacetolytic fragments and of positive ions of the b-series in CID-MS, the generation of intermediate oxazolonium ions in both gas and liquid phase is concluded.

Biological Variation of the Platelet Proteome in the Elderly Population and Its Implication for Biomarker Research

Knowledge about the extent of total variation experienced between samples from different individuals is of great importance for the design of not only proteomics but every clinical study. This variation defines the smallest statistically significant detectable signal difference when comparing two groups of individuals. We isolated platelets from 20 healthy human volunteers aged 56-100 years because this age group is most commonly encountered in the clinics. We determined the technical and total variation experienced in a proteome analysis using two-dimensional DIGE with IPGs in the pI ranges 4-7 and 6-9. Only spots that were reproducibly detectable in at least 90% of all gels (n = 908) were included in the study. All spots had a similar technical variation with a median coefficient of variation (cv) of about 7%. In contrast, spots showed a more diverse total variation between individuals with a surprisingly low median cv of only 18%. Because most known biomarkers show an effect size in a 1-2-fold range of their cv, any future clinical proteomics study with platelets will require an analytical method that is able to detect such small quantitative differences. In addition, we calculated the minimal number of samples (sample size) needed to detect given protein expression differences with statistical significance.

Synergy testing of multidrug resistant Acinetobacter baumanii against tigecycline and polymyxin using an E-test methodology

The emergence of multi drug resistant nosocomial Acinetobacter baumanii infection has prompted the evaluation of newer antimicrobial agents and older warehoused agents as possible therapeutic candidates. In addition to the carbapenems, two other agents have emerged in clinical use and the in-vitro literature as potential therapeutic agents. In a number of studies tigecycline, a new agent in the glycylcycline class, related to minocycline, and polymyxin B, a polypeptide antibiotic used parenterally in the past as a gram negative rod antimicrobial, have show promising activity against multidrug resistant A. baumanii isolates, including those isolates resistant to the carbapenems [1–3]. Tigecycline appears to be bacteriostatic in vitro and in one published clinical study there has been breakthrough bacteremia. Very little in vitro data exists on the activity of the combination of polymyxin plus tigecycline against multidrug resistant A. baumanii [4, 5]. The E-test methodology has proven reliable for in vitro antimicrobial sensitivity testing [6]. A modified E-test methodology for performing antimicrobial synergy tested has been preliminarily evaluated in the literature and although not extensively validated, has been found to correlate well with traditional checkerboard and time-kill methodologies in several studies [7, 8]. E-test synergy testing is performed by crossing the E-test strips for the 2 test antimicrobials at a 90° angle at their predetermined MIC value on an inoculated Mueller Hinton agar plate. The plates are then incubated for 24 h at 35°C. The MIC of each antimicrobial in combination is interpreted as the value at which the zone of inhibition intersects the E test strip. The fractional inhibitory concentration index (FICI) is calculated and interpreted for synergy, indifference or antagonism. The FICI=MICtigecycline in combination/MICtigecyclinealone+ MICpolymixin in combination/MICpolymixin alone. The FICI is interpreted as follows: 4.0 indicates antagonism. We tested 19 recent clinical isolates of multidrug resistant A. baumanii from a large university teaching hospital for susceptibility to tigecycline and polymyxin B and performed synergy testing on the combination using E-test methodology. Clinical sources of the isolates were as follows: respiratory 9/19 (47.4%), urine 6/19 (31.6%), blood 2/19 (10.5%), wound 2/19 (10.5%). E-test synergy testing, revealed FICI values for tigecycline plus polymyxin between 0.88 and 2.0 for all 19 isolates, with 11/19 isolates having an FICI of 2.0. The FICI distribution was as follows:

New Sequences, Constituents, and Producers of Peptaibiotics: An Updated Review

To date, 18 genera of imperfect and ascomycetous fungi have been recognized to produce ca. 700 individual sequences of peptaibiotics. These are linear polypeptide antibiotics which i) have a molecular weight between 500 and 2,200 Dalton, thus containing 5-21 residues; ii) show a high content of alpha-aminoisobutyric acid; iii) are characterized by the presence of other nonproteinogenic amino acids and/or lipoamino acids; iv) possess an acylated N-terminus, and v) have a C-terminal residue that, in most of them, consists of a free or acetylated amide-bonded 1,2-amino alcohol, but might also be an amine, amide, free amino acid, 2,5-dioxopiperazine, or sugar alcohol. From April 2003 until present, ca. 300 new individual sequences of peptaibiotics have been published in the literature, but most of them have not yet been included in databases. To summarize these new sequences and novel constituents, as well as to introduce fungal species hitherto unknown as producers of peptaibiotics, the relevant literature is reviewed. Furthermore, ecophysiological and taxonomic aspects of the producing fungi are discussed.

Peptaibiomics: Microheterogeneity, Dynamics, and Sequences of Trichobrachins, Peptaibiotics from Trichoderma parceramosumBissett (T. longibrachiatumRifai)

From the culture broth of the filamentous fungus Trichoderma parceramosum, strain CBS 936.69, a mixture of polypeptide antibiotics (pepaibiotics), named trichobrachin (TB), was isolated. Three major groups designated TB I, TB II, and TB III could be separated and isolated by preparative TLC on silica gel. Individual peptides of these three groups were sequenced by on-line LC/ESI-MS(n). The mixture of N-acetylated peptides comprises ten 19-residue peptides with a free C-terminal Gln residue (TB I peptides), two 18-residue peptides with a free C-terminal Gln residue (TB II 1 and 2), seven 20-residue peptides with a C-terminal amide-bound phenylalaninol (TB II 3-10), and 34 eleven-residue peptides with either a C-terminal leucinol or isoleucinol or valinol (TB III 1-34). Monitoring production and degradation of peptaibiotics in a pilot experiment revealed that the biosynthesis of TB II and TB III peptides starts two days after the beginning of fermentation. After five days of fermentation, the concentration of TB II decreased, whereas the amount of TB I increased. This observation unequivocally demonstrates that those two 18-residue TB I and TB II peptides with the free carboxy terminus result from enzymatic C-terminal degradation of the 20-residue TB II peptides. In analogy to the technical terms proteome and proteomics, the terms peptaibiome and peptaibiomics have recently been proposed for the entirety and dynamics of the Aib-containing peptides (comprehensively named peptaibiotics). Consequently, the entire peptaibiome of T. parceramosum grown under submerse conditions in shake-flasks for five days comprises at least 54 peptides differing in main-chain length and microheterogeneity, i.e., exchange of amino acids and the C-terminal 1,2-amino alcohol.