ORCID Profile
0000-0002-7091-9763
Current Organisation
The University of Newcastle
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Microbial genetics | Natural products and bioactive compounds | Microbiology | Synthetic biology |
Publisher: Springer Science and Business Media LLC
Date: 24-04-2023
DOI: 10.1007/S00248-023-02222-W
Abstract: Bacteria residing in the guts of pollinating insects play a key role in nutrient acquisition, digestion, and resistance to pests and diseases. Imbalances in microbial flora in response to environmental change and stress can therefore impact insect health and resilience. This study is aimed at defining the core gut microbiome of the Australian native stingless bee, Tetragonula carbonaria , and exploring the impact of colony transplantation on gut health. The gut microbiomes of nine forager bees from natural (log) and manufactured (box) hives were examined via 16S rRNA gene licon sequencing. Some differences were observed at the ASV level between the microbiomes of log and box hive bees. However, a core microbiome, dominated by Lactobacillus spp., unclassified Acetobacteraceae spp., and Bombella spp., was maintained. Further, the inferred functional potential of the microbiomes was consistent across all in iduals. This study highlights that although hive transplantation has an impact on the overall ersity of stingless bee gut microbiomes, it is unlikely to have a significant negative impact on the overall health and resilience of the colony.
Publisher: Elsevier BV
Date: 03-2012
Publisher: Elsevier BV
Date: 06-2008
DOI: 10.1016/J.COPBIO.2008.03.002
Abstract: Toxic cyanobacteria pose a significant hazard to human health and the environment. The recent characterisation of cyanotoxin synthetase gene clusters has resulted in an explosion of molecular detection methods for these organisms and their toxins. Conventional polymerase chain reaction (PCR) tests targeting cyanotoxin biosynthesis genes provide a rapid and sensitive means for detecting potentially toxic populations of cyanobacteria in water supplies. The adaptation of these simple PCR tests into quantitative methods has additionally enabled the monitoring of dynamic bloom populations and the identification of particularly problematic species. More recently, DNA microarray technology has been applied to cyanobacterial diagnostics offering a high-throughput option for detecting and differentiating toxic genotypes in complex s les. Together, these molecular methods are proving increasingly important for monitoring water quality.
Publisher: Wiley
Date: 26-06-2023
Abstract: Antimicrobial resistance (AMR) is predicted to cause a worldwide annual toll of 10 million deaths by 2050. This looming public health threat has been linked to antibiotic overuse and pollution, which places selective pressures on AMR maintenance and transfer in and between microbial populations. We examined the distribution, ersity and potential mobility of AMR genes in cyanobacteria. While cyanobacteria are not pathogenic, we hypothesised that they could be a major environmental reservoir for AMR genes. Genes encoding AMR to seven antimicrobial drug classes were found in 10% of cyanobacterial genomes. AMR genes were found in 13% of freshwater, 19% of terrestrial, 34% of symbiotic, 2% of thermal spring, and 3% of marine genomes. AMR genes were found in five cyanobacterial orders with 23% of Nostocales and 8% of Oscillatoriales strains containing AMR genes. The most frequently observed alleles were ansamycin resistance genes, which were present in 7% of strains. AMR genes responsible for resistance to broad‐spectrum β‐lactams, chlor henicols, tetracyclines, macrolides, and aminoglycosides were associated with mobile genetic elements or plasmid replicons or both. These results suggest that cyanobacteria are an extensive reservoir, and potential vector, for AMR genes in erse terrestrial and aquatic habitats.
Publisher: Informa UK Limited
Date: 2008
DOI: 10.1080/10408440802291513
Abstract: Over the last 10 years, we have witnessed major advances in our understanding of natural product biosynthesis, including the genetic basis for toxin production by numerous groups of cyanobacteria. Cyanobacteria produce an unparalleled array of bioactive secondary metabolites, including alkaloids, polyketides and non-ribosomal peptides, some of which are potent toxins. This review addresses the molecular genetics underlying the production of hepatotoxins, microcystin and nodularin in fresh and brackish water. These toxins pose a serious threat to human health and their occurrence in water supplies is increasing, because of the prevalence of toxic algal blooms worldwide. Toxin biosynthesis gene-cluster-associated transposition and the natural transformability of certain species suggest a broader distribution of toxic cyanobacterial taxa. The information gained from the discovery of these toxin biosynthetic pathways has enabled the genetic screening of various environments for drinking-water quality management. Understanding the role of cyanotoxins in the producing microorganisms and the environmental regulation of their biosynthesis genes may also suggest the means of controlling toxic-bloom events.
Publisher: Elsevier BV
Date: 2021
Publisher: Wiley
Date: 20-03-2013
DOI: 10.1111/J.1462-2920.2012.02729.X
Abstract: Over the past 15 years, the genetic basis for production of many cyanobacterial bioactive compounds has been described. This knowledge has enabled investigations into the environmental factors that regulate the production of these toxins at the molecular level. Such molecular or systems level studies are also likely to reveal the physiological role of the toxin and contribute to effective water resource management. This review focuses on the environmental regulation of some of the most relevant cyanotoxins, namely the microcystins, nodularin, cylindrospermopsin, saxitoxins, anatoxins and jamaicamides.
Publisher: Oxford University Press (OUP)
Date: 19-06-2013
Abstract: Metal homoeostasis in cyanobacteria is based on uptake and export systems that are controlled by their own regulators. This study characterises the zinc uptake (Znu) system in Nostoc punctiforme. The system was found to comprise of three subunits in an ACB operon: a Zn(2+)-binding protein (ZnuA18), a transmembrane domain (ZnuB) and an ATPase (ZnuC). These proteins are encoded within the znu operon regulated by a zinc uptake transcription repressor (Zur). Interestingly, a second Zn(2+)-binding protein (ZnuA08) was also identified at a distal genomic location. Interactions between components of the ZnuACB system were investigated using knockouts of the in idual genes. The znuA08(-), znuA18(-), znuB(-) and znuC(-) mutants displayed overall reduced znuACB transcript levels, suggesting that all system components are required for normal expression of znu genes. Zinc uptake assays in the Zn(2+)-binding protein mutant strains showed that the disruption of znuA18 had a greater negative effect on zinc uptake than disruption of znuA08. Complementation studies in Escherichia coli indicated that both znuA08 and znuA18 were able to restore zinc uptake in a znuA(-) mutant, with znuA18 permitting the highest zinc uptake rate. The N. punctiforme zur was also able to complement the E. coli zur(-) mutant.
Publisher: American Society for Microbiology
Date: 15-10-2016
DOI: 10.1128/AEM.01632-16
Abstract: Mycosporine-like amino acids (MAAs) are an important class of secondary metabolites known for their protection against UV radiation and other stress factors. Cyanobacteria produce a variety of MAAs, including shinorine, the active ingredient in many sunscreen creams. Bioinformatic analysis of the genome of the soil-dwelling cyanobacterium Cylindrospermum stagnale PCC 7417 revealed a new gene cluster with homology to MAA synthase from Nostoc punctiforme . This newly identified gene cluster is unusual because it has five biosynthesis genes ( mylA to mylE ), compared to the four found in other MAA gene clusters. Heterologous expression of mylA to mylE in Escherichia coli resulted in the production of mycosporine-lysine and the novel compound mycosporine-ornithine. To our knowledge, this is the first time these compounds have been heterologously produced in E. coli and structurally characterized via direct spectral guidance. This study offers insight into the ersity, biosynthesis, and structure of cyanobacterial MAAs and highlights their amenability to heterologous production methods. IMPORTANCE Mycosporine-like amino acids (MAAs) are significant from an environmental microbiological perspective as they offer microbes protection against a variety of stress factors, including UV radiation. The heterologous expression of MAAs in E. coli is also significant from a biotechnological perspective as MAAs are the active ingredient in next-generation sunscreens.
Publisher: American Society for Microbiology
Date: 08-2016
DOI: 10.1128/AEM.70.11.6370-6378.2004
Abstract: The cyanobacterium Microcystis aeruginosa is widely known for its production of the potent hepatotoxin microcystin. Microcystin is synthesized nonribosomally by the thiotemplate function of a large, modular enzyme complex encoded within the 55-kb microcystin synthetase ( mcy ) gene cluster. Also encoded within the mcy gene cluster is a putative ATP binding cassette (ABC) transporter, McyH. This study details the bioinformatic and mutational analyses of McyH and offers functional predictions for the hypothetical protein. The transporter is putatively comprised of two homodimers, each with an N-terminal hydrophobic domain and a C-terminal ATPase. Phylogenetically, McyH was found to cluster with members of the ABC-A 1 subgroup of ABC ATPases, suggesting an export function for the protein. Two mcyH null mutant (Δ mcyH ) strains were constructed by partial deletion of the mcyH gene. Microcystin production was completely absent in these strains. While the mcyH deletion had no apparent effect on the transcription of other mcy genes, the complete microcystin biosynthesis enzyme complex could not be detected in Δ mcyH mutant strains. Finally, expression levels of McyH in the wild type and in Δ mcyA , Δ mcyB , and Δ mcyH mutants were investigated by using immunoblotting with an anti-McyH antibody. Expression of McyH was found to be reduced in Δ mcyA and Δ mcyB mutants and completely absent in the Δ mcyH mutant. By virtue of its association with the mcy gene cluster and the bioinformatic and experimental data presented in this study, we predict that McyH functions as a microcystin exporter and is, in addition, intimately associated with the microcystin biosynthesis pathway.
Publisher: Wiley
Date: 28-05-2021
Publisher: Oxford University Press (OUP)
Date: 24-09-2015
DOI: 10.1111/JAM.12942
Abstract: To characterize genes involved in maintaining homeostatic levels of zinc in the cyanobacterium Nostoc punctiforme. Metal efflux transporters play a central role in maintaining homeostatic levels of trace elements such as zinc. Sequence analyses of the N. punctiforme genome identified two potential cation diffusion facilitator (CDF) metal efflux transporters, Npun_F0707 (Cdf31) and Npun_F1794 (Cdf33). Deletion of either Cdf31or Cdf33 resulted in increased zinc retention over 3 h. Interestingly, Cdf31(-) and Cdf33(-) mutants showed no change in sensitivity to zinc exposure in comparison with the wild type, suggesting some compensatory capacity for the loss of each other. Using qRT-PCR, a possible interaction was observed between the two cdf's, where the Cdf31(-) mutant had a more profound effect on cdf33 expression than Cdf33(-) did on cdf31. Over-expression of Cdf31 and Cdf33 in ZntA(-) - and ZitB(-) -deficient Escherichia coli revealed function similarities between the ZntA and ZitB of E. coli and the cyanobacterial transporters. The data presented shed light on the function of two important transporters that regulate zinc homeostasis in N. punctiforme. This study shows for the first time the functional characterization of two cyanobacterial zinc efflux proteins belonging to the CDF family.
Publisher: Springer Science and Business Media LLC
Date: 30-06-2013
DOI: 10.1007/S00253-013-5047-Y
Abstract: The ZIP family of metal transporters is involved in the transport of Zn(2+) and other metal cations from the extracellular environment and/or organelles into the cytoplasm of prokaryotes, eukaryotes and archaeotes. In the present study, we identified twin ZIP transporters, Zip11 (Npun_F3111) and Zip63 (Npun_F2202) encoded within the genome of the filamentous cyanobacterium, Nostoc punctiforme PCC73120. Sequence-based analyses and structural predictions confirmed that these cyanobacterial transporters belong to the SLC39 subfamily of metal transporters. Quantitative real-time (QRT)-PCR analyses suggested that the enzymes encoded by zip11 and zip63 have a broad allocrite range that includes zinc as well as cadmium, cobalt, copper, manganese and nickel. Inactivation of either zip11 or zip63 via insertional mutagenesis in N. punctiforme resulted in reduced expression of both genes, highlighting a possible co-regulation mechanism. Uptake experiments using (65)Zn demonstrated that both zip mutants had diminished zinc uptake capacity, with the deletion of zip11 resulting in the greatest overall reduction in (65)Zn uptake. Over-expression of Zip11 and Zip63 in an E. coli mutant strain (ZupT736::kan) restored alent metal cation uptake, providing further evidence that these transporters are involved in Zn uptake in N. punctiforme. Our findings show the functional role of these twin metal uptake transporters in N. punctiforme, which are independently expressed in the presence of an array of metals. Both Zip11 and Zip63 are required for the maintenance of homeostatic levels of intracellular zinc N. punctiforme, although Zip11 appears to be the primary zinc transporter in this cyanobacterium, both ZIP's may be part of a larger metal uptake system with shared regulatory elements.
Publisher: Elsevier BV
Date: 04-2012
DOI: 10.1016/J.WATRES.2011.12.019
Abstract: The bloom-forming, toxic cyanobacterium, Cylindrospermopsis raciborskii exhibits global distribution. In recent years both the occurrence and dominance of this species, particularly in temperate regions, has increased. Whilst this may be due to increased sensitivity of analytical detection methods or more rigorous s ling routines, it is possible that this expansion has been assisted by a number of changing conditions in these environments. The geographical expansion of both the organism and toxin production can be attributed to phenomena such as eutrophication and climate change. In this review, we discuss the occurrence of C. raciborskii with respect to current literature against the backdrop of increasing global temperatures. Critically, we identify a concerning trend between the geographical spread of this organism and global climate change.
Publisher: Royal Society of Chemistry (RSC)
Date: 2019
DOI: 10.1039/C8NP00063H
Abstract: This review discusses cyanotoxin biosynthetic pathways and highlights the heterologous expression and biochemical studies used to characterise them.
Publisher: CRC Press
Date: 25-03-2014
DOI: 10.1201/B16752-49
Publisher: Wiley
Date: 29-06-2022
Abstract: The soil surface of drylands can typically be colonized by cyanobacteria and other microbes, forming biological soil crusts or ‘biocrusts’. Biocrusts provide critical benefits to ecosystems and are a common component of the largely arid and semi‐arid Australian continent. Yet, their distribution and the parameters that shape their microbial composition have not been investigated. We present here the first detailed description of Australia's biocrust microbiome assessed from 15 sites across the continent using 16S rRNA sequencing. The most abundant bacterial phyla from all sites were Cyanobacteria, Proteobacteria, Actinobacteria, Chloroflexi and Bacteroidetes. Cyanobacterial communities from northern regions were more erse and unclassified cyanobacteria were a noticeable feature of northern biocrusts. Segregation between northern and southern regions was largely due to the differential abundance of Microcoleus spp., with M . paludosus dominating in the north and M . vaginatus dominating in the south. The geographical shifts in bacterial composition and ersity were correlated to seasonal temperatures and summer rainfall. Our findings provide an initial reference for s ling strategies to maximize access to bacterial genetic ersity. As hubs for essential ecosystem services, further investigation into biocrusts in arid and semi‐arid regions may yield discoveries of genetic mechanisms that combat increases in warming due to climate change.
Publisher: Wiley
Date: 15-02-2021
Publisher: Springer Science and Business Media LLC
Date: 29-01-2014
Publisher: Springer New York
Date: 2008
Publisher: CRC Press
Date: 03-2013
DOI: 10.1201/B13853
Publisher: Oxford University Press (OUP)
Date: 19-03-2021
DOI: 10.1093/GBE/EVAB056
Abstract: Cyanobacteria are prolific producers of natural products, including polyketides and hybrid compounds thereof. Type III polyketide synthases (PKSs) are of particular interest, due to their wide substrate specificity and simple reaction mechanism, compared with both type I and type II PKSs. Surprisingly, only two type III PKS products, hierridins, and (7.7)paracyclophanes, have been isolated from cyanobacteria. Here, we report the mining of 517 cyanobacterial genomes for type III PKS biosynthesis gene clusters. Approximately 17% of the genomes analyzed encoded one or more type III PKSs. Together with already characterized type III PKSs, the phylogeny of this group of enzymes was investigated. Our analysis showed that type III PKSs in cyanobacteria evolved into three major lineages, including enzymes associated with 1) (7.7)paracyclophane-like biosynthesis gene clusters, 2) hierridin-like biosynthesis gene clusters, and 3) cytochrome b5 genes. The evolutionary history of these enzymes is complex, with some sequences partitioning primarily according to speciation and others putatively according to their reaction type. Protein modeling showed that cyanobacterial type III PKSs generally have a smaller active site cavity (mean = 109.035 Å3) compared with enzymes from other organisms. The size of the active site did not correlate well with substrate size, however, the “Gatekeeper” amino acid residues within the active site were strongly correlated to enzyme phylogeny. Our study provides unprecedented insight into the distribution, ersity, and molecular evolution of cyanobacterial type III PKSs, which could facilitate the discovery, characterization, and exploitation of novel enzymes, biochemical pathways, and specialized metabolites from this biosynthetically talented clade of microorganisms.
Publisher: American Society for Microbiology
Date: 26-02-2021
DOI: 10.1128/AEM.02604-20
Abstract: This study demonstrates that the Pseudoalteromonas strain HM-SA03, isolated from the venomous blue-ringed octopus, Hapalochalaena sp., is a biosynthetically talented organism, capable of producing alterochromides and potentially six other specialized metabolites. We identified a pseudoalterobactin biosynthesis gene cluster and proposed a pathway for the production of the associated siderophore.
Publisher: Elsevier BV
Date: 08-2021
Publisher: Elsevier BV
Date: 02-2007
Publisher: Oxford University Press (OUP)
Date: 11-06-2021
DOI: 10.1111/JAM.15122
Publisher: American Society for Microbiology
Date: 07-2010
DOI: 10.1128/AEM.01862-09
Abstract: NtcA is a transcription factor that has been found in a erse range of cyanobacteria. This nitrogen-controlled factor was focused on as a key component in the yet-to-be-deciphered regulatory network controlling microcystin production. Adaptor-mediated PCR was utilized to isolate the ntcA gene from Microcystis aeruginosa PCC 7806. This gene was cloned, and the recombinant (His-tagged) protein was overexpressed and purified for use in mobility shift assays to analyze NtcA binding to putative sites identified in the microcystin mcyA / D promoter region. Autoregulation of NtcA in M. aeruginosa was shown via NtcA binding in the upstream ntcA promoter region. The observation of binding of NtcA to the mcyA / D promoter region has direct relevance for the regulation of microcystin biosynthesis, as transcription of the mcyABCDEFGHIJ gene cluster appears to be under direct control of nitrogen.
Publisher: American Society for Microbiology
Date: 12-04-2022
DOI: 10.1128/AEM.02373-21
Abstract: Iron availability limits the growth of many microorganisms, particularly those residing in high nutrient–low chlorophyll aquatic environments. Therefore, characterizing iron acquisition pathways in phytoplankton is essential for understanding nutrient cycling in our oceans.
Publisher: Wiley
Date: 16-11-2020
Abstract: Paralytic shellfish toxins (PSTs) are neurotoxic alkaloids produced by freshwater cyanobacteria and marine dinoflagellates. Due to their antagonism of voltage‐gated sodium channels in excitable cells, certain analogues are of significant pharmacological interest. The biosynthesis of the parent compound, saxitoxin, is initiated with the formation of 4‐amino‐3‐oxo‐guanidinoheptane (ethyl ketone) by an unusual polyketide synthase‐like enzyme, SxtA. We have heterologously expressed SxtA from Raphidiopsis raciborskii T3 in Escherichia coli and analysed its activity in vivo . Ethyl ketone and a truncated analogue, methyl ketone, were detected by HPLC‐ESI‐HRMS analysis, thus suggesting that SxtA has relaxed substrate specificity in vivo . The chemical structures of these products were further verified by tandem mass spectrometry and labelled‐precursor feeding with [guanidino‐ 15 N 2 ] arginine and [1,2‐ 13 C 2 ] acetate. These results indicate that the reactions catalysed by SxtA could give rise to multiple PST variants, including analogues of ecological and pharmacological significance.
Publisher: CSIRO Publishing
Date: 2020
DOI: 10.1071/MF18406
Abstract: The cyanobacterium Microcystis aeruginosa has been linked to toxic blooms worldwide. In addition to producing hepatotoxic microcystins, many strains are capable of synthesising a variety of biologically active compounds, including protease and phosphatase inhibitors, which may affect aquatic ecosystems and pose a risk to their use. This study explored the distribution, composition and conservation of known secondary metabolite (SM) biosynthesis gene clusters in the genomes of 27 M. aeruginosa strains isolated from six different Köppen–Geiger climates. Our analysis identified gene clusters with significant homology to nine SM biosynthesis gene clusters spanning four different compound classes: non-ribosomal peptides, hybrid polyketide–non-ribosomal peptides, cyanobactins and microviridins. The aeruginosin, microviridin, cyanopeptolin and microcystin biosynthesis gene clusters were the most frequently observed, but hybrid polyketide–non-ribosomal peptide biosynthesis clusters were the most common class overall. Although some biogeographic relationships were observed, taxonomic markers and geography were not reliable indicators of SM biosynthesis cluster distribution, possibly due to previous genetic deletions or horizontal gene transfer events. The only cyanotoxin biosynthesis gene cluster identified in our screening study was the microcystin synthetase (mcy) gene cluster, suggesting that the production of non-microcystin cyanotoxins by this taxon, such as anatoxin-a or paralytic shellfish poison analogues, is either absent or rare.
Publisher: Georg Thieme Verlag KG
Date: 07-2008
Publisher: CRC Press
Date: 29-10-2018
DOI: 10.1201/B22030
Publisher: CRC Press
Date: 23-04-2009
Publisher: Elsevier BV
Date: 04-2016
DOI: 10.1016/J.HAL.2015.11.002
Abstract: The production of toxic metabolites by cyanobacterial blooms represents a significant threat to the health of humans and ecosystems worldwide. Here we summarize the current state of the knowledge regarding the genetics, biosynthesis and regulation of well-characterized cyanotoxins, including the microcystins, nodularin, cylindrospermopsin, saxitoxins and anatoxins, as well as the lesser-known marine toxins (e.g. lyngbyatoxin, aplysiatoxin, jamaicamides, barbamide, curacin, hectochlorin and apratoxins).
Publisher: MDPI AG
Date: 10-05-2010
DOI: 10.3390/MD8051650
Publisher: American Chemical Society (ACS)
Date: 05-2019
DOI: 10.1021/ACSSYNBIO.9B00068
Abstract: The microcystins are a large group of cyclic peptide hepatotoxins produced by several genera of freshwater cyanobacteria. The genes responsible for microcystin biosynthesis are encoded within a large (∼55 kbp) gene cluster, mcyA-J. The recent establishment of a cyanotoxin heterologous expression system in Escherichia coli has provided the means to study microcystin biosynthesis in a genetically tractable, rapidly growing host. Using this system, we demonstrate that deletion of the ABC-transporter, mcyH, and dehydrogenase, mcyI, abolishes microcystin production, while deletion of the O-methyltransferase, mcyJ, results in the production of the demethylated (DM) toxin [d-Asp
Publisher: Wiley
Date: 21-03-2022
DOI: 10.1111/GBI.12489
Abstract: Microbialites are sedimentary rocks created in association with benthic microorganisms. While they harbour complex microbial communities, Cyanobacteria perform critical roles in sediment stabilisation and accretion. Microbialites have been described from permanent and ephemeral saline lakes in South Australia however, the microbial communities that generate and inhabit these biogeological structures have not been studied in detail. To address this knowledge gap, we investigated the composition, ersity and metabolic potential of bacterial communities from different microbialite‐forming mats and surrounding sediments in five South Australian saline coastal lakes using 16S rRNA gene sequencing and predictive metagenome analyses. While Proteobacteria and Bacteroidetes were the dominant phyla recovered from the mats and sediments, Cyanobacteria were significantly more abundant in the mat s les. Interestingly, at lower taxonomic levels, the mat communities were vastly different across the five lakes. Comparative analysis of putative mat and sediment metagenomes via PICRUSt2 revealed important metabolic pathways driving the process of carbonate precipitation, including cyanobacterial oxygenic photosynthesis, ureolysis and nitrogen fixation. These pathways were highly conserved across the five examined lakes, although they appeared to be performed by distinct groups of bacterial taxa found in each lake. Stress response, quorum sensing and circadian clock were other important pathways predicted by the in silico metagenome analysis. The enrichment of CRISPR/Cas and phage shock associated genes in these cyanobacteria‐rich communities suggests that they may be under selective pressure from viral infection. Together, these results highlight that a very stable ecosystem function is maintained by distinctly different communities in microbialite‐forming mats in the five South Australian lakes and reinforce the concept that ‘who’ is in the community is not as critical as their net metabolic capacity.
Start Date: 2023
End Date: 12-2025
Amount: $630,000.00
Funder: Australian Research Council
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