scholarly journals A non-canonical peptide synthetase adenylates 3-methyl-2-oxovaleric acid for auriculamide biosynthesis

2016 ◽  
Vol 12 ◽  
pp. 2766-2770 ◽  
Author(s):  
Daniel Braga ◽  
Dirk Hoffmeister ◽  
Markus Nett
Keyword(s):  
Structural Variation ◽  
Genetic Locus ◽  
Functional Characterization ◽  
Structural Diversity ◽  
Building Blocks ◽  
Adenylation Domain ◽  
Proteinogenic Amino Acid ◽  
Keto Acids ◽  
Peptide Synthetase

Auriculamide is the first natural product known from the predatory bacteriumHerpetosiphon aurantiacus.It is composed of three unusual building blocks, including the non-proteinogenic amino acid 3-chloro-L-tyrosine, the α-hydroxy acid L-isoleucic acid, and a methylmalonyl-CoA-derived ethane unit. A candidate genetic locus for auriculamide biosynthesis was identified and encodes four enzymes. Among them, the non-canonical 199 kDa four-domain nonribosomal peptide synthetase, AulA, is extraordinary in that it features two consecutive adenylation domains. Here, we describe the functional characterization of the recombinantly produced AulA. The observed activation of 3-methyl-2-oxovaleric acid by the enzyme supports the hypothesis that it participates in the biosynthesis of auriculamide. An artificially truncated version of AulA that lacks the first adenylation domain activated this substrate like the full-length enzyme which shows that the first adenylation domain is dispensable. Additionally, we provide evidence that the enzyme tolerates structural variation of the substrate. α-Carbon substituents significantly affected the substrate turnover. While all tested aliphatic α-keto acids were accepted by the enzyme and minor differences in chain size and branches did not interfere with the enzymatic activity, molecules with methylene α-carbons led to low turnover. Such enzymatic plasticity is an important attribute to help in the perpetual search for novel molecules and to access a greater structural diversity by mutasynthesis.

Coordinated Action of NRPS, Baeyer-Villiger Monooxygenase, and Methyltransferase Ensures the Economical Biosynthesis of Bohemamines in Streptomyces sp. CB02009

2020 ◽  
Author(s):  
Ling Liu ◽  
Sainan Li ◽  
Runze Sun ◽  
Xiangjing Qin ◽  
Jianhua Ju ◽  
...  
Keyword(s):  
Methyl Groups ◽  
Ribosome Engineering ◽  
Methyl Group ◽  
Proteinogenic Amino Acid ◽  
Streptomyces Sp ◽  
Coordinated Action ◽  
Peptide Synthetase ◽  
Identification And Characterization ◽  
Shed Light

<p> Bohemamines (BHMs) are bacterial alkaloids containing a pyrrolizidine core with two unprecedented methyl groups. Herein we report the activation of BHMs biosynthesis in <i>Streptomyces </i>sp. CB02009 using a ribosome engineering approach. Identification and characterization of the <i>bhm</i> gene cluster reveals a coordinated action of nonribosomal peptide synthetase BhmJ, Baeyer-villiger monooxygenase BhmK and methyltransferase BhmG for BHMs biosynthesis. BhmG is responsible for the C-methylation on C-7, while the C-9 methyl group is from a non-proteinogenic amino acid (2<i>S</i>,5<i>S</i>)-5-methylproline, required for BHMs production in three model <i>Streptomyces </i>hosts. Our study shed light on the intricate interaction of BhmJ/BhmK/BhmG for the economical biosynthesis of BHMs in their native producer, and also unraveled that BhmJ and BhmK are competent biocatalysts in <i>S</i><i>treptomyce </i><i>albus</i>.</p>

Functional Characterization of Compliant Building Blocks Utilizing Eigentwists and Eigenwrenches

2008 ◽  
Author(s):  
Charles J. Kim
Keyword(s):  
Building Block ◽  
Compliant Mechanisms ◽  
Compliant Mechanism ◽  
Deformable Body ◽  
Functional Characterization ◽  
Building Blocks ◽  
Future Research ◽  
Special Cases ◽  
Block Approach

Compliant mechanisms are devices which utilize the flexibility of their constituent members to transmit motion and forces. Unlike their rigid body counterparts, compliant mechanisms typically contain no traditional joints. The focus of this research is the development of a building block approach for the synthesis of compliant mechanisms. Building block methods better facilitate the augmentation of designer intuition while offering a systematic approach to open-ended problems. In this paper, we investigate the use of the eigentwists and eigenwrenches of a deformable body to characterize basic kinematic function. The eigentwists and eigenwrenches are shown to demonstrate parametric behavior when applied to the compliant dyad building block, and in special cases may be compared to compliance ellipsoids. The paper concludes by articulating future research in a building block approach to compliant mechanism synthesis.

scholarly journals Plasticity and Evolution of Aeruginosin Biosynthesis in Cyanobacteria

2009 ◽  
Vol 75 (7) ◽  
pp. 2017-2026 ◽  
Author(s):  
Keishi Ishida ◽  
Martin Welker ◽  
Guntram Christiansen ◽  
Sabrina Cadel-Six ◽  
Christiane Bouchier ◽  
...  
Keyword(s):  
Molecular Basis ◽  
Common Ancestor ◽  
Structural Diversity ◽  
Gene Clusters ◽  
Adenylation Domain ◽  
Peptide Synthetase ◽  
Structural Peculiarities ◽  
Tailoring Enzymes ◽  
The Individual ◽  
Insight Into

ABSTRACT Aeruginosins are bioactive oligopeptides that are produced in high structural diversity by strains of the bloom-forming cyanobacterial genera Microcystis and Planktothrix. A hallmark of aeruginosins is the unusual Choi moiety central to the tetrapeptides, while other positions are occupied by variable moieties in individual congeners. Here we report on three aeruginosin synthetase gene clusters (aer) of Microcystis aeruginosa (strains PCC 7806, NIES-98, and NIES-843). The analysis and comparison the aer gene clusters provide the first insight into the molecular basis of biosynthetic and structural plasticity in aeruginosin pathways. Major parts of the aer gene clusters are highly similar in all strains, particularly the genes coding for the first three nonribosomal peptide synthetase (NRPS) modules except for the region coding for the second adenylation domain. However, the gene clusters differ largely in genes coding for tailoring enzymes such as halogenases and sulfotransferases, reflecting structural peculiarities in aeruginosin congeners produced by the individual strains. Significant deviations were further observed in the C-terminal NRPS modules, suggesting two distinct release mechanisms. The architecture of the gene clusters is in agreement with the particular aeruginosin variants that are produced by individual strains, the structures of two of which (aeruginosins 686 A and 686 B) were elucidated. The aer gene clusters of Microcystis and Planktothrix are proposed to originate from a common ancestor and to have evolved to their present-day diversity largely through horizontal gene transfer and recombination events.

scholarly journals Physical and functional characterization of the genetic locus of IBtk, an inhibitor of Bruton's tyrosine kinase: evidence for three protein isoforms of IBtk

2008 ◽  
Vol 36 (13) ◽  
pp. 4402-4416 ◽  
Author(s):  
Carmen Spatuzza ◽  
Marco Schiavone ◽  
Emanuela Di Salle ◽  
Elzbieta Janda ◽  
Marco Sardiello ◽  
...  
Keyword(s):  
Tyrosine Kinase ◽  
Genetic Locus ◽  
Functional Characterization ◽  
Protein Isoforms ◽  
Bruton’S Tyrosine Kinase ◽  
Bruton's Tyrosine Kinase

Coordinated Action of NRPS, Baeyer-Villiger Monooxygenase, and Methyltransferase Ensures the Economical Biosynthesis of Bohemamines in Streptomyces sp. CB02009

2020 ◽  
Author(s):  
Ling Liu ◽  
Sainan Li ◽  
Runze Sun ◽  
Xiangjing Qin ◽  
Jianhua Ju ◽  
...  
Keyword(s):  
Methyl Groups ◽  
Ribosome Engineering ◽  
Methyl Group ◽  
Proteinogenic Amino Acid ◽  
Streptomyces Sp ◽  
Coordinated Action ◽  
Peptide Synthetase ◽  
Identification And Characterization ◽  
Shed Light

<p> Bohemamines (BHMs) are bacterial alkaloids containing a pyrrolizidine core with two unprecedented methyl groups. Herein we report the activation of BHMs biosynthesis in <i>Streptomyces </i>sp. CB02009 using a ribosome engineering approach. Identification and characterization of the <i>bhm</i> gene cluster reveals a coordinated action of nonribosomal peptide synthetase BhmJ, Baeyer-villiger monooxygenase BhmK and methyltransferase BhmG for BHMs biosynthesis. BhmG is responsible for the C-methylation on C-7, while the C-9 methyl group is from a non-proteinogenic amino acid (2<i>S</i>,5<i>S</i>)-5-methylproline, required for BHMs production in three model <i>Streptomyces </i>hosts. Our study shed light on the intricate interaction of BhmJ/BhmK/BhmG for the economical biosynthesis of BHMs in their native producer, and also unraveled that BhmJ and BhmK are competent biocatalysts in <i>S</i><i>treptomyce </i><i>albus</i>.</p>

scholarly journals Platyhelminth Venom Allergen-Like (VAL) proteins: revealing structural diversity, class-specific features and biological associations across the phylum

Parasitology ◽  
2012 ◽  
Vol 139 (10) ◽  
pp. 1231-1245 ◽  
Author(s):  
IAIN W. CHALMERS ◽  
KARL F. HOFFMANN
Keyword(s):  
Immune Response ◽  
Functional Characterization ◽  
Structural Diversity ◽  
Host Immune Response ◽  
Comprehensive Analysis ◽  
Protein Family ◽  
Host Interactions ◽  
Venom Allergen

SUMMARYDuring platyhelminth infection, a cocktail of proteins is released by the parasite to aid invasion, initiate feeding, facilitate adaptation and mediate modulation of the host immune response. Included amongst these proteins is the Venom Allergen-Like (VAL) family, part of the larger sperm coating protein/Tpx-1/Ag5/PR-1/Sc7 (SCP/TAPS) superfamily. To explore the significance of this protein family during Platyhelminthes development and host interactions, we systematically summarize all published proteomic, genomic and immunological investigations of the VAL protein family to date. By conducting new genomic and transcriptomic interrogations to identify over 200 VAL proteins (228) from species in all 4 traditional taxonomic classes (Trematoda, Cestoda, Monogenea and Turbellaria), we further expand our knowledge related to platyhelminth VAL diversity across the phylum. Subsequent phylogenetic and tertiary structural analyses reveal several class-specific VAL features, which likely indicate a range of roles mediated by this protein family. Our comprehensive analysis of platyhelminth VALs represents a unifying synopsis for understanding diversity within this protein family and a firm context in which to initiate future functional characterization of these enigmatic members.

Functional characterization of human brown adipose tissue metabolism

2020 ◽  
Vol 477 (7) ◽  
pp. 1261-1286 ◽  
Author(s):  
Marie Anne Richard ◽  
Hannah Pallubinsky ◽  
Denis P. Blondin
Keyword(s):  
Adipose Tissue ◽  
Brown Adipose Tissue ◽  
Functional Characterization ◽  
Human Infants ◽  
Positron Emission ◽  
Brown Adipose ◽  
Treatment Of Obesity ◽  
And Function

Brown adipose tissue (BAT) has long been described according to its histological features as a multilocular, lipid-containing tissue, light brown in color, that is also responsive to the cold and found especially in hibernating mammals and human infants. Its presence in both hibernators and human infants, combined with its function as a heat-generating organ, raised many questions about its role in humans. Early characterizations of the tissue in humans focused on its progressive atrophy with age and its apparent importance for cold-exposed workers. However, the use of positron emission tomography (PET) with the glucose tracer [18F]fluorodeoxyglucose ([18F]FDG) made it possible to begin characterizing the possible function of BAT in adult humans, and whether it could play a role in the prevention or treatment of obesity and type 2 diabetes (T2D). This review focuses on the in vivo functional characterization of human BAT, the methodological approaches applied to examine these features and addresses critical gaps that remain in moving the field forward. Specifically, we describe the anatomical and biomolecular features of human BAT, the modalities and applications of non-invasive tools such as PET and magnetic resonance imaging coupled with spectroscopy (MRI/MRS) to study BAT morphology and function in vivo, and finally describe the functional characteristics of human BAT that have only been possible through the development and application of such tools.

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