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Complete genome sequence of Streptomyces lividans TK24

Streptomyces lividans TK24 is the standard host for the heterologous expression of a number of different proteins and antibiotic-synthesizing enzymes. As such, it is often used as an experimental microbial cell factory for the production of secreted heterologous proteins including human cytokines and industrial enzymes, and of several antibiotics. It accepts methylated DNA and is an ideal Streptomyces cloning system. Here, we report the complete genome sequence of S. lividans TK24 that includes a plasmid-less genome of 8.345 Mbp (72.24% G + C content).

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Biochemical characterization and validation of a catalytic site of a highly thermostable Ts2631 endolysin from the Thermus scotoductus phage vB_Tsc2631

Phage vB_Tsc2631 infects the extremophilic bacterium Thermus scotoductus MAT2631 and uses the Ts2631 endolysin for the release of its progeny. The Ts2631 endolysin is the first endolysin from thermophilic bacteriophage with an experimentally validated catalytic site. In silico analysis and computational modelling of the Ts2631 endolysin structure revealed a conserved Zn2+ binding site (His30, Tyr58, His131 and Cys139) similar to Zn2+ binding site of eukaryotic peptidoglycan recognition proteins (PGRPs). We have shown that the Ts2631 endolysin lytic activity is dependent on divalent metal ions (Zn2+ and Ca2+). The Ts2631 endolysin substitution variants H30N, Y58F, H131N and C139S dramatically lost their antimicrobial activity, providing evidence for the role of the aforementioned residues in the lytic activity of the enzyme. The enzyme has proven to be not only thermoresistant, retaining 64.8% of its initial activity after 2 h at 95°C, but also highly thermodynamically stable (Tm = 99.82°C, ΔHcal = 4.58 × 104 cal mol-1). Substitutions of histidine residues (H30N and H131N) and a cysteine residue (C139S) resulted in variants aggregating at temperatures ≥75°C, indicating a significant role of these residues in enzyme thermostability. The substrate spectrum of the Ts2631 endolysin included extremophiles of the genus Thermus but also Gram-negative mesophiles, such as Escherichia coliSalmonella panamaPseudomonas fluorescens and Serratia marcescens. The broad substrate spectrum and high thermostability of this endolysin makes it a good candidate for use as an antimicrobial agent to combat Gram-negative pathogens.

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Skýrslur

Pigmentation Trial with Whiteleg Shrimp

Útgefið:

03/06/2021

Höfundar:

Wolfgang Koppe, Georges Lamborelle

Tengiliður

Georges Lamborelle

Stöðvarstjóri tilraunaeldisstöðvar

georges@matis.is

This report presents the results of a feeding experiment performed by Matis ohf. for Adisseo. The trial objective was to compare the effect of Asta-S on the intensity of pigmentation in whiteleg shrimp (Penaeus vannamei), compared with two other pigmenting feed additives (Carophyll Pink and Panaferd). The diets were formulated by Dr. Wolfgang Koppe (Matis) and produced in the feed production facilities of Matis as pellets suitable for adult P. vannamei. The trial was carried out at the Matís Aquaculture Research Station (MARS). A 24-day feeding period was done in triplicate tanks with 10 shrimp each. The average temperature was at 26.35 °C and salinity at 22.3 ppt.

Skýrslan er lokuð / This report is closed

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Stock structure of Atlantic herring Clupea harengus in the Norwegian Sea and adjacent waters

The genetic structure of Atlantic herring Clupea harengus L. was investigated in its north-easterly distribution in the Norwegian Sea and adjacent waters, using 23 neutral and one non-neutral (Cpa111) microsatellite loci. Fish from the suspected 2 main populations—the Norwegian spring-spawning herring (NSSH) and the Icelandic summer-spawning herring (ISSH)—were collected at spawning locations in their respective spawning seasons from 2009 to 2012. Samples were also collected from Norwegian autumn spawning locations, from different local Norwegian fjords such as the inner part of Trondheimsfjorden, Lindås pollene, Landvikvannet and Lusterfjorden, as well as from suspected Faroese spawning components. The observed level of genetic differentiation was significant but low (FST = 0.007) and mostly attributable to the differentiation of the local Norwegian fjord populations. The locus Cpa111, which was detected to putatively be under positive selection, exhibited the highest FST value (0.044). The observed genetic patterns were robust to exclusion of this locus. Landvikvannet herring was also genetically distinguishable from the 3 other fjord populations. In addition, the present study does not support genetic structuring among the ISSH and the NSSH.

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Rhodopsin Gene Polymorphism Associated with Divergent Light Environments in Atlantic Cod

The spectral sensitivity of visual pigments in vertebrate eyes is optimized for specific light conditions. One of such pigments, rhodopsin (RH1), mediates dim-light vision. Amino acid replacements at tuning sites may alter spectral sensitivity, providing a mechanism to adapt to ambient light conditions and depth of habitat in fish. Here we present a first investigation of RH1 gene polymorphism among two ecotypes of Atlantic cod in Icelandic waters, which experience divergent light environments throughout the year due to alternative foraging behaviour. We identified one synonymous single nucleotide polymorphism (SNP) in the RH1 protein coding region and one in the 3′ untranslated region (3′-UTR) that are strongly divergent between these two ecotypes. Moreover, these polymorphisms coincided with the well-known panthophysin (Pan I) polymorphism that differentiates coastal and frontal (migratory) populations of Atlantic cod. While the RH1 SNPs do not provide direct inference for a specific molecular mechanism, their association with this dim-sensitive pigment indicates the involvement of the visual system in local adaptation of Atlantic cod.

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Innflutt fersk matvæli og sýkingaráhætta fyrir menn

Aðgangur að öruggum matvælum er hluti af forréttindum Íslendinga. Hratt vaxandi sýklalyfjaónæmi, sem meðal annars er tengt verksmiðjubúum og mikilli sýklalyfjanotkun í landbúnaði, er ein helsta ógn við lýðheilsu mannkyns. Vaxandi verslun með matvæli á milli landa og heimsálfa auðveldar dreifingu sýkla og sýklalyfjaónæmis um heiminn. Íslenskur landbúnaður og landbúnaðarafurðir hafa sérstöðu vegna einangrunar landsins og smæðar.

Eftir umfangsmiklar aðgerðir til að draga úr útbreiðslu Campylobacter og Salmonella á kjúklingabúum er nýgengi Campylobactersýkinga orðið um 17-43/100.000 íbúa, þar af helmingurinn af innlendum uppruna, og Salmonellusýkinga 10-15/100.000, og þar af smitast flestir í útlöndum. Enterohaemorrhagic E. coli (EHEC) hefur ekki fundist í íslenskum nautgripum og kemur lágt nýgengi (að jafnaði 0-0,6/100.000) því ekki á óvart. Nýleg hópsýking af völdum fjölónæms EHEC-stofns var rakin til innflutts mengaðs salats. Sýklalyfjanotkun í íslenskum landbúnaði er ein sú lægsta sem þekkist í Evrópu og fágætt er að innlent smit af völdum Salmonella og Campylobacter sé af völdum sýklalyfjaónæmra stofna. Karbapenemasa-myndandi Enterobacteriaceae hafa enn ekki fundist á Íslandi.

Lítil notkun sýklalyfja í íslenskum landbúnaði ásamt aðhaldsaðgerðum til að draga úr útbreiðslu Campylobacter og Salmonella hafa borið mikinn árangur. Almenningur þarf að vera meðvitaður um mikilvægi uppruna matvæla og að íslenskar landbúnaðarafurðir hafa enn sérstöðu með tilliti til smithættu.

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Chondroitin Lyase from a Marine Arthrobacter sp. MAT3885 for the Production of Chondroitin Sulfate Disaccharides

Chondroitin sulfate (CS) saccharides from cartilage tissues have potential application in medicine or as dietary supplements due to their therapeutic bioactivities. Studies have shown that depolymerized CS saccharides may display enhanced bioactivity. The objective of this study was to isolate a CS-degrading enzyme for an efficient production of CS oligo- or disaccharides. CS-degrading bacteria from marine environments were enriched using in situ artificial support colonization containing CS from shark cartilage as substrate. Subsequently, an Arthrobacter species (strain MAT3885) efficiently degrading CS was isolated from a CS enrichment culture. The genomic DNA from strain MAT3885 was pyro-sequenced by using the 454 FLX sequencing technology. Following assembly and annotation, an orf, annotated as family 8 polysaccharide lyase genes, was identified, encoding an amino acid sequence with a similarity to CS lyases according to NCBI blastX. The gene, designated choA1, was cloned in Escherichia coli and expressed downstream of and in frame with the E. coli malE gene for obtaining a high yield of soluble recombinant protein. Applying a dual-tag system (MalE-Smt3-ChoA1), the MalE domain was separated from ChoA1 with proteolytic cleavage using Ulp1 protease. ChoA1 was defined as an AC-type enzyme as it degraded chondroitin sulfate A, C, and hyaluronic acid. The optimum activity of the enzyme was at pH 5.5–7.5 and 40 °C, running a 10-min reaction. The native enzyme was estimated to be a monomer. As the recombinant chondroitin sulfate lyase (designated as ChoA1R) degraded chondroitin sulfate efficiently compared to a benchmark enzyme, it may be used for the production of chondroitin sulfate disaccharides for the food industry or health-promoting products.

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The effects of pre-salting methods on salt and water distribution of heavily salted cod, as analyzed by H-1 and Na-23, Na-23 NMR, low-field NMR and physicochemical analysis

The effect of different pre-salting methods (brine injection with salt with/without polyphosphates, brining and pickling) on the water and salt distribution in dry salted Atlantic cod (Gadus morhua) fillets was studied with proton and sodium NMR and MRI methods, supported by physicochemical analysis of salt and water content as well as water holding capacity. The study indicated that double head brine injection with salt and phosphates lead to the least heterogeneous water distribution, while pickle salting had the least heterogeneous salt distribution. Fillets from all treatments contained spots with unsaturated brine, increasing the risk of microbial denaturation of the fillets during storage. Since a homogeneous water and salt distribution was not achieved with the studied pre-salting methods, further optimizations of the salting process, including the pre-salting and dry salting steps, must be made in the future.

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Gene-associated markers can assign origin in a weakly structured fish, Atlantic herring

Regulations on the exploitation of populations of commercially important fish species and the ensuing consumer interest in sustainable products have increased the need to accurately identify the population of origin of fish and fish products. Although genomics-based tools have proven highly useful, there are relatively few examples in marine fish displaying accurate origin assignment. We synthesize data for 156 single-nucleotide polymorphisms typed in 1039 herring, Clupea harengus L., spanning the Northeast Atlantic to develop a tool that allows assignment of individual herring to their regional origin. We show the method’s suitability to address specific biological questions, as well as management applications. We analyse temporally replicated collections from two areas, the Skagerrak (n = 81, 84, 66) and the western Baltic (n = 52, 52). Both areas harbour heavily fished mixed-origin stocks, complicating management issues. We report novel genetic evidence that herring from the Baltic Sea contribute to catches in the North Sea, and find support that western Baltic feeding aggregations mainly constitute herring from the western Baltic with contributions from the Eastern Baltic. Our study describes a general approach and outlines a database allowing individual assignment and traceability of herring across a large part of its East Atlantic distribution.

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A CGTase with high coupling activity using {gamma}-cyclodextrin isolated from a novel strain clustering under the genus Carboxydocella

Cyclodextrin glucanotransferases (CGTases; EC 2.4.1.19) have mainly been characterized for their ability to produce cyclodextrins (CDs) from starch in an intramolecular transglycosylation reaction (cyclization). However, this class of enzymes can also catalyze intermolecular transglycosylation via disproportionation or coupling reactions onto a wide array of acceptors and could therefore be valuable as a tool for glycosylation.In this paper, we report the gene isolation, via the CODEHOP strategy, expression and characterization of a novel CGTase (CspCGT13) from a Carboxydocella sp. This enzyme is the first glycoside hydrolase isolated from the genus, indicating starch degradation via cyclodextrin production in the Carboxydocella strain. The fundamental reactivities of this novel CGTase are characterized and compared with two commercial CGTases, assayed under identical condition, in order to facilitate interpretation of the results. The comparison showed that the enzyme, CspCGT13, displayed high coupling activity using γ-CD as donor, despite preferentially forming α- and β-CD in the cyclization reaction using wheat starch as substrate. Comparison of subsite conservation within previously characterized CGTases showed significant sequence variation in subsites −3 and −7, which may be important for the coupling activity.

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