Module Extension of a Non‐Ribosomal Peptide Synthetase of the Glycopeptide Antibiotic Balhimycin Produced by <i>Amycolatopsis balhimycina</i>

Butz, Diane; Schmiederer, Timo; Hadatsch, Bianka; Wohlleben, Wolfgang; Weber, Tilmann; Süssmuth, Roderich D.

doi:10.1002/cbic.200800068

Cited by 48 publications

(37 citation statements)

References 23 publications

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“…Manipulation of their modular organization provides enormous potential for generating novel bioactive compounds (Miao et al, 2006; Butz et al, 2008; Doekel et al, 2008). Matching pairs of COM domains form protein–protein interactions (Linne et al, 2003; Hahn and Stachelhaus, 2006), and the present study demonstrated the decisive role of COM domain pairs in NRP biosynthetic complexes.…”

Section: Discussionmentioning

confidence: 99%

Biocombinatorial Synthesis of Novel Lipopeptides by COM Domain-Mediated Reprogramming of the Plipastatin NRPS Complex

et al. 2016

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Both donors and acceptors of communication-mediating (COM) domains are essential for coordinating intermolecular communication within nonribosomal peptides synthetases (NRPSs) complexes. Different sets of COM domains provide selectivity, allowing NRPSs to utilize different natural biosynthetic templates. In this study, novel lipopeptides were synthesized by reprogramming the plipastatin biosynthetic machinery. A Thr-to-Asp point mutation was sufficient to shift the selectivity of the donor COM domain of ppsB toward that of ppsD. Deletion and/or interchangeability established donor and acceptor function. Variations in acceptor COM domain did not result in novel product formation in the presence of its partner donor, whereas plipastatin formation was completely abrogated by altering donor modules. Five novel lipopeptides (cyclic pentapeptide, linear hexapeptide, nonapeptide, heptapeptide, and cyclic octapeptide) were identified and verified by high-resolution LC-ESI-MS/MS. In addition, we demonstrated the potential to generate novel strains with the antimicrobial activity by selecting compatible COM domains, and the novel lipopeptides exhibited antimicrobial activity against five of the fungal species at a contention of 31.25–125 μg/ml.

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Section: Discussionmentioning

confidence: 99%

Biocombinatorial Synthesis of Novel Lipopeptides by COM Domain-Mediated Reprogramming of the Plipastatin NRPS Complex

et al. 2016

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“…Amino acid insertion by module extension has been performed within the vancomycin-type glycopeptide antibiotic, balhimycin. Insertion of an entire d -hydroxyphenylglycine module into the balhimycin assembly line between modules 4 and 5 results in an elongated octapeptide product [163]. C -terminally truncated hexa- to di-peptide metabolites are also detected.…”

Section: Genetic Engineering Of Nrps To Create Novel Productsmentioning

confidence: 99%

Diversity of Nonribosomal Peptide Synthetases Involved in the Biosynthesis of Lipopeptide Biosurfactants

Roongsawang

Washio

Morikawa

2010

IJMS

210

164

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Lipopeptide biosurfactants (LPBSs) consist of a hydrophobic fatty acid portion linked to a hydrophilic peptide chain in the molecule. With their complex and diverse structures, LPBSs exhibit various biological activities including surface activity as well as anti-cellular and anti-enzymatic activities. LPBSs are also involved in multi-cellular behaviors such as swarming motility and biofilm formation. Among the bacterial genera, Bacillus (Gram-positive) and Pseudomonas (Gram-negative) have received the most attention because they produce a wide range of effective LPBSs that are potentially useful for agricultural, chemical, food, and pharmaceutical industries. The biosynthetic mechanisms and gene regulation systems of LPBSs have been extensively analyzed over the last decade. LPBSs are generally synthesized in a ribosome-independent manner with megaenzymes called nonribosomal peptide synthetases (NRPSs). Production of active-form NRPSs requires not only transcriptional induction and translation but also post-translational modification and assemblage. The accumulated knowledge reveals the versatility and evolutionary lineage of the NRPSs system. This review provides an overview of the structural and functional diversity of LPBSs and their different biosynthetic mechanisms in Bacillus and Pseudomonas, including both typical and unique systems. Finally, successful genetic engineering of NRPSs for creating novel lipopeptides is also discussed.

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“…Module deletions (Mootz et al, 2002), insertions (Butz et al, 2008), and fusions of unrelated modules (Duerfahrt et al, 2003;Mootz et al, 2000) identified additional degrees of freedom for combinatorial biosynthesis. Mechanistic investigations of individual domains helped to circumvent unproductive module combinations (Belshaw et al, 1999).…”

Section: Introductionmentioning

confidence: 99%

A Subdomain Swap Strategy for Reengineering Nonribosomal Peptides

Kries¹,

Niquille²,

Hilvert³

2015

Chemistry & Biology

133

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Nonribosomal peptide synthetases (NRPSs) protect microorganisms from environmental threats by producing diverse siderophores, antibiotics, and other peptide natural products. Their modular molecular structure is also attractive from the standpoint of biosynthetic engineering. Here we evaluate a methodology for swapping module specificities of these mega-enzymes that takes advantage of flavodoxin-like subdomains involved in substrate recognition. Nine subdomains encoding diverse specificities were transplanted into the Phe-specific GrsA initiation module of gramicidin S synthetase. All chimeras could be purified as soluble protein. One construct based on a Val-specific subdomain showed sizable adenylation activity and functioned as a Val-Pro diketopiperazine synthetase upon addition of the proline-specific GrsB1 module. These results suggest that subdomain swapping could be a viable alternative to previous NRPS design approaches targeting binding pockets, domains, or entire modules. The short length of the swapped sequence stretch may facilitate straightforward exploitation of the wealth of existing NRPS modules for combinatorial biosynthesis.

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Module Extension of a Non‐Ribosomal Peptide Synthetase of the Glycopeptide Antibiotic Balhimycin Produced by Amycolatopsis balhimycina

Cited by 48 publications

References 23 publications

Biocombinatorial Synthesis of Novel Lipopeptides by COM Domain-Mediated Reprogramming of the Plipastatin NRPS Complex

Biocombinatorial Synthesis of Novel Lipopeptides by COM Domain-Mediated Reprogramming of the Plipastatin NRPS Complex

Diversity of Nonribosomal Peptide Synthetases Involved in the Biosynthesis of Lipopeptide Biosurfactants

A Subdomain Swap Strategy for Reengineering Nonribosomal Peptides

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