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Rapid Communications
in Mass Spectrometry
Mar 2006; 20(8): 1176 - 1180
http://dx.doi.org/10.1002/rcm.2430
© 2006 John Wiley & Sons, Ltd.
Archimer, archive institutionnelle de l’Ifremer
http://www.ifremer.fr/docelec/
New short peptaibols from a marine Trichoderma strain
Mustapha Mohamed-Benkada
1
, Monique Montagu
1
*, Jean-François Biard
1
,
Florence Mondeguer
2
, Philippe Verite
3
, Michèle Dalgalarrondo
4
,
John Bissett
5
, Yves François Pouchus
1
1
Université de Nantes, Groupe SMAB-EA 2160, Faculté de pharmacie, BP 53508, 44035 Nantes Cedex 1, France
2
IFREMER- DEL/PN, 1 Rue de l'Ile d'Yeu Nantes BP 21105, 44 311 Nantes Cedex 3, France
3
UFR mixte Médecine-Pharmacie, Laboratoire de Chimie Analytique, 22 boulevard Gambetta, 76183 Rouen
France
4
LEIMA-INRA, rue de la Géraudière, BP 71627, 44316 Nantes Cedex 3, France
5
Agriculture and Agri-Food Canada, ECORC, Neatby Bldg. 21, 960 Carling Av., Ottawa, Ontario, Canada K1A
OC6
*: Corresponding author : Monique.Montagu@univ-nantes.fr
Abstract: The production of peptaibols by a marine-related Trichoderma longibrachiatum strain was
studied using electrospray ionisation multiple-stage ion trap mass spectrometry (ESI-MSn-IT) and gas
chromatography/electron impact mass spectrometry (GC/EI-MS). Two major groups of peptaibols
were identified, those with long sequences (20 amino acids) and others with short sequences (11
amino acids). This paper describes the methodology used to establish the sequences of short
peptaibols in a mixture without previous individual separation. Nine peptaibols were identified. Among
them, eight are new, namely as trichobrachin A I-IV (Aib9-Pro10 sequence) and as trichobrachin B I-IV
(Val9-Pro10 sequence). Original Pro6-Val7 and Val9-Pro10 sequences have to be noted. Copyright ©
2006 John Wiley & Sons, Ltd.
Keywords: Short peptaibols, electrospray ionisation-multiple-stage mass spectrometry-ion trap (ESI-
MSn-IT),marine products, marine fungi, Trichoderma longibrachiatum
For Peer Review
New short peptaibols from a marine Trichoderma strain
Journal: Rapid Communications in Mass Spectrometry
Manuscript ID: RCM-05-1071.R1
Wiley - Manuscript type: Research Article
Date Submitted by the
Author:
n/a
Complete List of Authors: MONTAGU, Monique; Université de Nantes, Groupe SMAB EA2160
Faculté de Pharmacie
Keywords:
Short peptaibols, Trichoderma longibrachiatum, marine fungi,
marine products, electrospray ionisation-multiple-stage mass
spectrometry-ion trap
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Rapid Communications in Mass Spectometry
For Peer Review
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New short peptaibols from a marine Trichoderma strain1
2
3
Mustapha MOHAMED-BENKADA
1
, Monique MONTAGU
1*
, Jean-François BIARD
1
, Florence 4
MONDEGUER
2
, Philippe VERITE
3
, Michèle DALGALARRONDO
4
, John BISSETT
5
, Yves François 5
POUCHUS
1
6
7
1– Université de Nantes - Groupe SMAB-EA 2160 - Faculté de pharmacie - BP 53508 - 44035 Nantes 8
Cedex 1- France *tel. : +33 2-40-41-28-65, fax : +33 2-40-41-28-58, 9
2- IFREMER- DEL/ PN, 1, Rue de l’Ile d’Yeu Nantes B.P. 21105, 44 311 Nantes Cedex 3, France.10
3- UFR mixte Médecine-Pharmacie, Laboratoire de Chimie Analytique, 22 boulevard Gambetta 76183 11
Rouen France. 12
4- LEIMA-INRA- rue de la Géraudière BP 71627 44316 Nantes Cedex 3 France. 13
5- Agriculture and Agri-Food Canada, ECORC, Neatby Bldg. 21, 960 Carling Av., Ottawa, Ontario, 14
Canada K1A OC6.15
16
17
*Corresponding author: M. Montagu, Université de Nantes - Groupe SMAB-EA 2160 - Faculté de 18
pharmacie - BP 53508 - 44035 Nantes Cedex 1, France19
email: Monique.Montagu@univ-nantes.fr20
Tel: +33 2-40-41-28-65 21
Fax: +33 2-40-41-28-5822
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ABSTRACT23
24
The production of peptaibols by a marine-related Trichoderma longibrachiatum strain was studied 25
using electrospray ionisation-multiple-stage mass spectrometry-ion trap (ESI-MS
n
-IT) and gas 26
chromatography electron impact mass spectrometry (GC/EI-MS). Two major groups of peptaibols were 27
identified, those with long sequences (20 amino-acids) and others with short sequences (11 amino-28
acids). This paper describes the methodology used to establish the sequences of short peptaibols in a 29
mixture without previous individual separation. Nine peptaibols were identified. Among them, eight are 30
new, namely as trichobrachin A I-IV (Aib
9
-Pro
10
sequence) and as trichobrachin B I-IV (Val
9
-Pro
10
31
sequence). Original Pro
6
-Val
7
and Val
9
-Pro
10
sequences have to be noted. 32
33
KEY WORDS34
35
Short peptaibols, electrospray ionisation-multiple-stage mass spectrometry-ion trap (ESI-MS
n
-IT), 36
marine products, marine fungi, Trichoderma longibrachiatum37
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INTRODUCTION 38
39
Peptaibols are an important class of linear peptides specific to fungi. The genus Trichoderma 40
(teleomorph Hypocrea: Ascomycota) is the most prolific known producer of peptaibols.
1,2
41
Approximately 200 of more than 300 known peptaibols have been identified from this genus alone.
3
42
Species of the genus Trichoderma are widespread in the marine environment and have been shown to 43
be able to produce peptaibols in marine culture conditions.
4,5
Peptaibols are characterized by a high 44
content of an uncommon amino acid:
-aminoisobutyric acid (Aib), an N-terminal acyl (most often 45
acetyl) group, and a C-terminal
-amino alcohol. According to the number (7 to 20) and the nature of 46
the amino acid (AA) sequence, they have been classified in 9 subfamilies (SF’s).
6
47
Short peptaibols produced by Trichoderma species belong to subfamily SF4. They are generally 48
constituted of 11 residues with 2 Pro at positions 6 and 10. Their sequences are microheterogeneous 49
analogues with different amino acids (AAs) in positions 2, 3, 4 and 11 (Figure 1a). There are about 50
forty short peptaibol sequences listed in the peptaibol database
7
including pseudokonins KL III and KL 51
VI produced by T. pseudokoningii, trichorovins by T. harzianum
1,8,9
and trikoningins KA and KB by T. 52
koningii.
10
Trichogin A IV is the unique short peptaibol described from T. longibrachiatum characterized 53
by a different elementary composition in AAs and especially by the absence of Pro residues in its 54
sequence (Figure 1b).
8
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Short peptaibols have been little studied. They are known to interact with double-layered phospholipid 56
membranes in the same manner as longer peptaibols. They form ion channels probably by using 57
unique insertion and conductance mechanisms. It has been supposed that, due to their short 58
sequences, they can span only the half of the lipid bilayer and two molecules have to be associated by 59
the N-terminal ends at the centre of the membrane.
6
Short peptaibols exhibit antibacterial (Gram
+
), 60
antifungal and antimycoplasmic activities.
1,8
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This paper describes the production, the purification and the sequence identification of new short 62
peptaibols produced by a marine strain of Trichoderma longibrachiatum Rifai. Microheterogeneous 63
sequence determination employed an original method with ESI-MS
n
-IT based on the establishment of a 64
filiation graph. Among the nine short peptaibols identified in this study, eight sequences are new. The 65
strain studied also produced several different long chain peptaibols with 20 AA residues which will be 66
analysed using the same protocol and reported in a separate communication.67
68
MATERIAL AND METHODS69
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