Microcystinase – a review of the natural occurrence, heterologous expression, and biotechnological application of MlrA

Dexter, Jason; McCormick, Alistair J.; Fu, Pengcheng; Dziga, Dariusz

doi:10.1016/j.watres.2020.116646

Cited by 23 publications

(16 citation statements)

References 112 publications

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“…(19). However, the presence of biodegradable hepatoxins MCs in Sphingomonadaceae has been reported more frequently (15). Through cloning and gene library screening, the gene clusters for biodegrading MC-LR (mlr A, mlr B, mlr C and mlr D) were identified preliminarily by Bourne et al (20).…”

Section: Discussionmentioning

confidence: 99%

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Genome sequencing and functional genes comparison betweenSphingopyxisUSTB-05 andSphingomonas morindaeNBD5

Liu¹,

Xu²,

Zhao³

et al. 2021

Preprint

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Sphingomonadaceae has a large number of strains that can biodegrade hepatotoxins or environmental pollutants. The latest research reported that certain strains can also produce lutein. Based on the third-generation sequencing technology, we analyzed the whole genome sequence and compared related functional genes of two strains of Sphingomonadaceae isolated from different habitats. The genome of Sphingopyxis USTB-05 was 4,679,489 bp and contained 4312 protein coding genes. The 4,239,716 bp nuclear genome of Sphingomonas morindae NBD5, harboring 3882 protein coding genes, has two sets of chromosomes. Both strains had lutein synthesis metabolism pathway sharing identical synthetic genes of crtB, crtE, crtI, crtQ, crtL, crtR, atoB, dxs, dxr, ispD, ispE, ispDF, gcpE, ispG, ispH, ispA, ispB and ispU. Sphingopyxis USTB-05 had hepatotoxins microcystins and nodularin metabolic pathways related to 16 genes (ald, ansA, gdhA, crnA, phy, ocd, hypdh, spuC, nspC, speE, murI, murD, murC, hmgL, bioAand glsA), while these genes were not found in Sphingomonas morindae NBD5. The unique protein sequences of strain NBD5 and strain USTB-05 were 155 and 199, respectively. The analysis of whole genome of the two Sphingomonadaceae strains provides insights into prokaryote evolution, the new pathway for lutein production and the new genes for environmental pollutant biodegradation.

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Section: Discussionmentioning

confidence: 99%

“…Nowadays, the mlr gene cluster encoding functional hydrolase have been identified in many hepatotoxin biodegrading bacteria (15). The mlr gene cluster contained mlrC, mlrA, mlrB and mlrD, corresponding to the enzymes MlrC, MlrA, MlrB and MlrD that biodegraded hepatotoxin.…”

Section: Comparison Of the Pathways And Genes Of Hepatotoxin Biodegradationmentioning

confidence: 99%

Genome sequencing and functional genes comparison betweenSphingopyxisUSTB-05 andSphingomonas morindaeNBD5

Liu¹,

Xu²,

Zhao³

et al. 2021

Preprint

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“…have been testified to contain MC-biodegrading gene cluster mlrABCD, which encoded MlrABCD enzymes (Bourne et al, 1996;Park et al, 2001;Ishii et al, 2004;Yan et al, 2012;Yang et al, 2020;Dexter et al, 2021). These enzymes play an important role in biodegrading MCs (Bourne et al, 1996(Bourne et al, , 2001(Bourne et al, , 2006Saito et al, 2003;Dexter et al, 2021). It has been demonstrated that quorum sensing systems positively regulated the degradation of MC by the transcriptional induction of these MC-degrading genes, especially mlrA (Zeng et al, 2020).…”

Section: Introductionmentioning

confidence: 99%

Characterization and Mechanism of Linearized-Microcystinase Involved in Bacterial Degradation of Microcystins

et al. 2021

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Microcystins (MCs) are extremely hazardous to the ecological environment and public health. How to control and remove MCs is an unsolved problem all over the world. Some microbes and their enzymes are thought to be effective in degrading MCs. Microcystinase can linearize microcystin-leucine-arginine (MC-LR) via a specific locus. However, linearized MC-LR is also very toxic and needs to be removed. How linearized MC-LR was metabolized by linearized-microcystinase, especially how linearized-microcystinase binds to linearized MC-LR, has not been defined. A combination of in vitro experiments and computer simulation was applied to explore the characterization and molecular mechanisms for linearized MC-LR degraded by linearized-microcystinase. The purified linearized-microcystinase was obtained by recombinant Escherichia coli overexpressing. The concentration of linearized MC-LR was detected by high-performance liquid chromatography, and linearized MC-LR degradation products were analyzed by the mass spectrometer. Homology modeling was used to predict the structure of the linearized-microcystinase. Molecular docking techniques on the computer were used to simulate the binding sites of linearized-microcystinase and linearized MC-LR. The purified linearized-microcystinase was obtained successfully. The linearized-microcystinase degraded linearized MC-LR to tetrapeptide efficiently. The second structure of linearized-microcystinase consisted of many alpha-helices, beta-strands, and colis. Linearized-microcystinase interacted the linearized MC-LR with hydrogen bond, hydrophobic interaction, electrostatic forces, and the Van der Waals force. This study firstly reveals the characterization and specific enzymatic mechanism of linearized-microcystinase for catalyzing linearized MC-LR. These findings encourage the application of MC-degrading engineering bacteria and build a great technique for MC-LR biodegradation in environmental engineering.

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“…Regarding their occurrence in the environment, when MCs are released into the water, they can remain relatively stable, regardless of environmental conditions, with half-life values ranging from less than 24 h to 15 days [16][17][18][19]. This fact might be influenced by their initial concentration and microbial community involved in biodegradation [16,[19][20][21]. Conversely, there is no clear information about the fate of other cyanopeptides in water systems, since monitoring is difficult due to the lack of standard materials [6,9,10,13].…”

Section: Introductionmentioning

confidence: 99%

“…The exploitation of biodegrading bacteria has been increasing, since they are seen as a safe and low-cost alternative compared to conventional or physical-chemical approaches for the remediation of contaminated water with cyanobacterial compounds [16,21]. Regarding MC biodegradation, many researchers have been contributing to the knowledge of isolated bacteria or microbial communities on the dynamics of this process in different conditions that can be applied as a water treatment process [16][17][18][19][21][22][23]. Although many techniques have been proposed to remove cyanotoxins from water, there are few reports about the application of a permanent and biotechnological system using viable microorganisms as a treatment in situ.…”

Section: Introductionmentioning

confidence: 99%

Degradation of Multiple Peptides by Microcystin-Degrader Paucibacter toxinivorans (2C20)

Santos

Soldatou

Magalhães

et al. 2021

Toxins

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Since conventional drinking water treatments applied in different countries are inefficient at eliminating potentially toxic cyanobacterial peptides, a number of bacteria have been studied as an alternative to biological filters for the removal of microcystins (MCs). Here, we evaluated the degradation of not only MCs variants (-LR/DM-LR/-RR/-LF/-YR), but also non-MCs peptides (anabaenopeptins A/B, aerucyclamides A/D) by Paucibactertoxinivorans over 7 days. We also evaluated the degradation rate of MC-LR in a peptide mix, with all peptides tested, and in the presence of M. aeruginosa crude extract. Furthermore, biodegradation was assessed for non-cyanobacterial peptides with different chemical structures, such as cyclosporin A, (Glu1)-fibrinopeptide-B, leucine-enkephalin, and oxytocin. When cyanopeptides were individually added, P. toxinivorans degraded them (99%) over 7 days, except for MC-LR and -RR, which decreased by about 85 and 90%, respectively. The degradation rate of MC-LR decreased in the peptide mix compared to an individual compound, however, in the presence of the Microcystis extract, it was degraded considerably faster (3 days). It was noted that biodegradation rates decreased in the mix for all MCs while non-MCs peptides were immediately degraded. UPLC–QTOF–MS/MS allowed us to identify two linear biodegradation products for MC-LR and MC-YR, and one for MC-LF. Furthermore, P. toxinivorans demonstrated complete degradation of non-cyanobacterial peptides, with the exception of oxytocin, where around 50% remained after 7 days. Thus, although P. toxinivorans was previously identified as a MC-degrader, it also degrades a wide range of peptides under a range of conditions, which could be optimized as a potential biological tool for water treatment.

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Microcystinase – a review of the natural occurrence, heterologous expression, and biotechnological application of MlrA

Cited by 23 publications

References 112 publications

Genome sequencing and functional genes comparison betweenSphingopyxisUSTB-05 andSphingomonas morindaeNBD5

Genome sequencing and functional genes comparison betweenSphingopyxisUSTB-05 andSphingomonas morindaeNBD5

Characterization and Mechanism of Linearized-Microcystinase Involved in Bacterial Degradation of Microcystins

Degradation of Multiple Peptides by Microcystin-Degrader Paucibacter toxinivorans (2C20)

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