Displaying publications 1 - 20 of 32 in total

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  1. Fulltext The Utilisation of Antarctic Microalgae Isolated from Paradise Bay (Antarctic Peninsula) in the Bioremediation of Diesel
    Zamree ND, Puasa NA, Lim ZS, Wong CY, Shaharuddin NA, Zakaria NN, et al.
    Plants (Basel), 2023 Jul 03;12(13).
    PMID: 37447097 DOI: 10.3390/plants12132536
    Research has confirmed that the utilisation of Antarctic microorganisms, such as bacteria, yeasts and fungi, in the bioremediation of diesel may provide practical alternative approaches. However, to date there has been very little attention towards Antarctic microalgae as potential hydrocarbon degraders. Therefore, this study focused on the utilisation of an Antarctic microalga in the bioremediation of diesel. The studied microalgal strain was originally obtained from a freshwater ecosystem in Paradise Bay, western Antarctic Peninsula. When analysed in systems with and without aeration, this microalgal strain achieved a higher growth rate under aeration. To maintain the growth of this microalga optimally, a conventional one-factor-at a-time (OFAT) analysis was also conducted. Based on the optimized parameters, algal growth and diesel degradation performance was highest at pH 7.5 with 0.5 mg/L NaCl concentration and 0.5 g/L of NaNO3 as a nitrogen source. This currently unidentified microalga flourished in the presence of diesel, with maximum algal cell numbers on day 7 of incubation in the presence of 1% v/v diesel. Chlorophyll a, b and carotenoid contents of the culture were greatest on day 9 of incubation. The diesel degradation achieved was 64.5% of the original concentration after 9 days. Gas chromatography analysis showed the complete mineralisation of C7-C13 hydrocarbon chains. Fourier transform infrared spectroscopy analysis confirmed that strain WCY_AQ5_3 fully degraded the hydrocarbon with bioabsorption of the products. Morphological and molecular analyses suggested that this spherical, single-celled green microalga was a member of the genus Micractinium. The data obtained confirm that this microalga is a suitable candidate for further research into the degradation of diesel in Antarctica.
  2. Discovery of microcystin-producing Anagnostidinema pseudacutissimum from cryopreserved Antarctic cyanobacterial mats
    Zaki S, Merican F, Muangmai N, Convey P, Broady P
    Harmful Algae, 2020 03;93:101800.
    PMID: 32307064 DOI: 10.1016/j.hal.2020.101800
    Microcystins (MCs) are secondary metabolites produced by cyanobacteria and have been well-documented in temperate and tropical regions. However, knowledge of the production of MCs in extremely cold environments is still in its infancy. Recently, examination of 100-year-old Antarctic cyanobacterial mats collected from Ross Island and the McMurdo Ice Shelf during Captain R.F. Scott's Discovery Expedition revealed that the presence of MCs in Antarctica is not a new phenomenon. Here, morphological and molecular phylogenetic analyses are used to identify a new microcystin-producing freshwater cyanobacterium, Anagnostidinema pseudacutissimum. The strain was isolated from a deep-frozen (-15 °C) sample collected from a red-brown cyanobacterial mat in a frozen pond at Cape Crozier (Ross Island, continental Antarctica) in 1984-1985. Amplification of the mcyE gene fragment involved in microcystin biosynthesis from A. pseudacutissimum confirmed that it is identical to the sequence from other known microcystin-producing cyanobacteria. Analysis of extracts from this A. pseudacutissimum strain by HPLC-MS/MS confirmed the presence of MC-LR and -YR at concentrations of 0.60 μg/L and MC-RR at concentrations of 0.20 μg/L. This is the first report of microcystin production from a species of Anagnostidinema from Antarctica.
  3. Fulltext Statistical Optimisation of Diesel Biodegradation at Low Temperatures by an Antarctic Marine Bacterial Consortium Isolated from Non-Contaminated Seawater
    Zakaria NN, Gomez-Fuentes C, Abdul Khalil K, Convey P, Roslee AFA, Zulkharnain A, et al.
    Microorganisms, 2021 Jun 03;9(6).
    PMID: 34205164 DOI: 10.3390/microorganisms9061213
    Hydrocarbon pollution is widespread around the globe and, even in the remoteness of Antarctica, the impacts of hydrocarbons from anthropogenic sources are still apparent. Antarctica's chronically cold temperatures and other extreme environmental conditions reduce the rates of biological processes, including the biodegradation of pollutants. However, the native Antarctic microbial diversity provides a reservoir of cold-adapted microorganisms, some of which have the potential for biodegradation. This study evaluated the diesel hydrocarbon-degrading ability of a psychrotolerant marine bacterial consortium obtained from the coast of the north-west Antarctic Peninsula. The consortium's growth conditions were optimised using one-factor-at-a-time (OFAT) and statistical response surface methodology (RSM), which identified optimal growth conditions of pH 8.0, 10 °C, 25 ppt NaCl and 1.5 g/L NH4NO3. The predicted model was highly significant and confirmed that the parameters' salinity, temperature, nitrogen concentration and initial diesel concentration significantly influenced diesel biodegradation. Using the optimised values generated by RSM, a mass reduction of 12.23 mg/mL from the initial 30.518 mg/mL (4% (w/v)) concentration of diesel was achieved within a 6 d incubation period. This study provides further evidence for the presence of native hydrocarbon-degrading bacteria in non-contaminated Antarctic seawater.
  4. Fulltext Mathematical Modelling of Canola Oil Biodegradation and Optimisation of Biosurfactant Production by an Antarctic Bacterial Consortium Using Response Surface Methodology
    Zahri KNM, Khalil KA, Gomez-Fuentes C, Zulkharnain A, Sabri S, Convey P, et al.
    Foods, 2021 Nov 14;10(11).
    PMID: 34829082 DOI: 10.3390/foods10112801
    An Antarctic soil bacterial consortium (reference BS14) was confirmed to biodegrade canola oil, and kinetic studies on this biodegradation were carried out. The purpose of this study was to examine the ability of BS14 to produce biosurfactants during the biodegradation of canola oil. Secondary mathematical equations were chosen for kinetic analyses (Monod, Haldane, Teissier-Edwards, Aiba and Yano models). At the same time, biosurfactant production was confirmed through a preliminary screening test and further optimised using response surface methodology (RSM). Mathematical modelling demonstrated that the best-fitting model was the Haldane model for both waste (WCO) and pure canola oil (PCO) degradation. Kinetic parameters including the maximum degradation rate (μmax) and maximum concentration of substrate tolerated (Sm) were obtained. For WCO degradation these were 0.365 min-1 and 0.308%, respectively, while for PCO they were 0.307 min-1 and 0.591%, respectively. The results of all preliminary screenings for biosurfactants were positive. BS14 was able to produce biosurfactant concentrations of up to 13.44 and 14.06 mg/mL in the presence of WCO and PCO, respectively, after optimisation. The optimum values for each factor were determined using a three-dimensional contour plot generated in a central composite design, where a combination of 0.06% salinity, pH 7.30 and 1.55% initial substrate concentration led to the highest biosurfactant production when using WCO. Using PCO, the highest biosurfactant yield was obtained at 0.13% salinity, pH 7.30 and 1.25% initial substrate concentration. This study could help inform the development of large-scale bioremediation applications, not only for the degradation of canola oil but also of other hydrocarbons in the Antarctic by utilising the biosurfactants produced by BS14.
  5. Fulltext The Use of Response Surface Methodology as a Statistical Tool for the Optimisation of Waste and Pure Canola Oil Biodegradation by Antarctic Soil Bacteria
    Zahri KNM, Zulkharnain A, Gomez-Fuentes C, Sabri S, Abdul Khalil K, Convey P, et al.
    Life (Basel), 2021 May 20;11(5).
    PMID: 34065265 DOI: 10.3390/life11050456
    Hydrocarbons can cause pollution to Antarctic terrestrial and aquatic ecosystems, both through accidental release and the discharge of waste cooking oil in grey water. Such pollutants can persist for long periods in cold environments. The native microbial community may play a role in their biodegradation. In this study, using mixed native Antarctic bacterial communities, several environmental factors influencing biodegradation of waste canola oil (WCO) and pure canola oil (PCO) were optimised using established one-factor-at-a-time (OFAT) and response surface methodology (RSM) approaches. The factors include salinity, pH, type of nitrogen and concentration, temperature, yeast extract and initial substrate concentration in OFAT and only the significant factors proceeded for the statistical optimisation through RSM. High concentration of substrate targeted for degradation activity through RSM compared to OFAT method. As for the result, all factors were significant in PBD, while only 4 factors were significant in biodegradation of PCO (pH, nitrogen concentration, yeast extract and initial substrate concentration). Using OFAT, the most effective microbial community examined was able to degrade 94.42% and 86.83% (from an initial concentration of 0.5% (v/v)) of WCO and PCO, respectively, within 7 days. Using RSM, 94.99% and 79.77% degradation of WCO and PCO was achieved in 6 days. The significant interaction for the RSM in biodegradation activity between temperature and WCO concentration in WCO media were exhibited. Meanwhile, in biodegradation of PCO the significant factors were between (1) pH and PCO concentration, (2) nitrogen concentration and yeast extract, (3) nitrogen concentration and PCO concentration. The models for the RSM were validated for both WCO and PCO media and it showed no significant difference between experimental and predicted values. The efficiency of canola oil biodegradation achieved in this study provides support for the development of practical strategies for efficient bioremediation in the Antarctic environment.
  6. Microcystin production by oscillatorialean cyanobacteria isolated from cryopreserved Antarctic mats
    Usman AS, Merican F, Zaki S, Broady P, Convey P, Muangmai N
    Harmful Algae, 2022 Dec;120:102336.
    PMID: 36470600 DOI: 10.1016/j.hal.2022.102336
    Twenty cyanobacterial strains of eight morphospecies isolated from deep-frozen (-15 °C) mat samples originally collected on Ross Island, in Victoria Land, and on the McMurdo Ice Shelf were screened for the presence of genes encoding for production of anatoxins, cylindrospermopsin, microcystin/nodularin and saxitoxin. One strain of each of Microcoleus autumnalis and Phormidesmis priestleyi and two strains of Wilmottia murrayi were found to produce microcystin. No toxin production was detected in the other 16 strains representing five species. The four toxin-producing strains were characterised using both morphological and molecular approaches. Phylogenetic analyses using partial 16S rRNA sequences were consistent with the morphological identification of all four strains. They were all found to contain a fragment of the mcyE gene, which is involved in microcystin biosynthesis. ELISA analysis of extracts from cultures of these strains confirmed the presence of low concentrations of microcystin: 0.35 μg/L in M. autumnalis, <0.15 μg/L in P. priestleyi, 1.60 μg/L in W. murrayi strain 1 and 0.9 μg/L in W. murrayi strain 2. This study includes the first report of microcystin synthesis by W. murrayi.
  7. Fulltext Strong and widespread cycloheximide resistance in Stichococcus-like eukaryotic algal taxa
    Syuhada NH, Merican F, Zaki S, Broady PA, Convey P, Muangmai N
    Sci Rep, 2022 Jan 20;12(1):1080.
    PMID: 35058560 DOI: 10.1038/s41598-022-05116-y
    This study was initiated following the serendipitous discovery of a unialgal culture of a Stichococcus-like green alga (Chlorophyta) newly isolated from soil collected on Signy Island (maritime Antarctica) in growth medium supplemented with 100 µg/mL cycloheximide (CHX, a widely used antibiotic active against most eukaryotes). In order to test the generality of CHX resistance in taxa originally identified as members of Stichococcus (the detailed taxonomic relationships within this group of algae have been updated since our study took place), six strains were studied: two strains isolated from recent substrate collections from Signy Island (maritime Antarctica) ("Antarctica" 1 and "Antarctica" 2), one isolated from this island about 50 years ago ("Antarctica" 3) and single Arctic ("Arctic"), temperate ("Temperate") and tropical ("Tropical") strains. The sensitivity of each strain towards CHX was compared by determining the minimum inhibitory concentration (MIC), and growth rate and lag time when exposed to different CHX concentrations. All strains except "Temperate" were highly resistant to CHX (MIC > 1000 µg/mL), while "Temperate" was resistant to 62.5 µg/mL (a concentration still considerably greater than any previously reported for algae). All highly resistant strains showed no significant differences in growth rate between control and treatment (1000 µg/mL CHX) conditions. Morphological examination suggested that four strains were consistent with the description of the species Stichococcus bacillaris while the remaining two conformed to S. mirabilis. However, based on sequence analyses and the recently available phylogeny, only one strain, "Temperate", was confirmed to be S. bacillaris, while "Tropical" represents the newly erected genus Tetratostichococcus, "Antarctica 1" Tritostichococcus, and "Antarctica 2", "Antarctica 3" and "Arctic" Deuterostichococcus. Both phylogenetic and CHX sensitivity analyses suggest that CHX resistance is potentially widespread within this group of algae.
  8. De novo assembly of complete genome sequence of Planococcus kocurii ATCC 43650(T), a potential plant growth promoting bacterium
    See-Too WS, Tan JY, Ee R, Lim YL, Convey P, Pearce DA, et al.
    Mar Genomics, 2016 Apr 22.
    PMID: 27117861 DOI: 10.1016/j.margen.2016.04.007
    Planococcus kocurii ATCC 43650(T) is a halotolerant and psychrotolerant bacterium isolated from the skin of a North sea cod. Here, we present the first complete genome and annotation of P. kocurii ATCC 43650(T), identifying its potential as a plant growth promoting bacterium and its capability in the biosynthesis of butanol.
  9. Complete genome of Pseudomonas sp. strain L10.10, a psychrotolerant biofertilizer that could promote plant growth
    See-Too WS, Lim YL, Ee R, Convey P, Pearce DA, Yin WF, et al.
    J Biotechnol, 2016 Mar 20;222:84-5.
    PMID: 26876481 DOI: 10.1016/j.jbiotec.2016.02.017
    Pseudomonas sp. strain L10.10 (=DSM 101070) is a psychrotolerant bacterium which was isolated from Lagoon Island, Antarctica. Analysis of its complete genome sequence indicates its possible role as a plant-growth promoting bacterium, including nitrogen-fixing ability and indole acetic acid (IAA)-producing trait, with additional suggestion of plant disease prevention attributes via hydrogen cyanide production.
  10. Complete genome of Planococcus rifietoensis M8(T), a halotolerant and potentially plant growth promoting bacterium
    See-Too WS, Convey P, Pearce DA, Lim YL, Ee R, Yin WF, et al.
    J Biotechnol, 2016 Mar 10;221:114-5.
    PMID: 26808870 DOI: 10.1016/j.jbiotec.2016.01.026
    Planococcus rifietoensis M8(T) (=DSM 15069(T)=ATCC BAA-790(T)) is a halotolerant bacterium with potential plant growth promoting properties isolated from an algal mat collected from a sulfurous spring in Campania (Italy). This paper presents the first complete genome of P. rifietoensis M8(T). Genes coding for various potentially plant growth promoting properties were identified within its genome.
  11. Complete genome sequence of Planococcus donghaensis JH1T, a pectin-degrading bacterium
    See-Too WS, Chua KO, Lim YL, Chen JW, Convey P, Mohd Mohidin TB, et al.
    J Biotechnol, 2017 Jun 20;252:11-14.
    PMID: 28483443 DOI: 10.1016/j.jbiotec.2017.05.005
    The type strain Planococcus donghaensis JH1Tis a psychrotolerant and halotolerant bacterium with starch-degrading ability. Here, we determine the carbon utilization profile of P. donghaensis JH1Tand report the first complete genome of the strain. This study revealed the strain's ability to utilize pectin and d-galacturonic acid, and identified genes responsible for degradation of the polysaccharides. The genomic information provided may serve as a fundamental resource for full exploration of the biotechnological potential of P. donghaensis JH1T.
  12. Pseudomonas versuta sp. nov., isolated from Antarctic soil
    See-Too WS, Salazar S, Ee R, Convey P, Chan KG, Peix Á
    Syst Appl Microbiol, 2017 Jun;40(4):191-198.
    PMID: 28501448 DOI: 10.1016/j.syapm.2017.03.002
    In this study we analysed three bacterial strains coded L10.10T, A4R1.5 and A4R1.12, isolated in the course of a study of quorum-quenching bacteria occurring in Antarctic soil. The 16S rRNA gene sequence was identical in the three strains and showed 99.7% pairwise similarity with respect to the closest related species Pseudomonas weihenstephanensis WS4993T. Therefore, the three strains were classified within the genus Pseudomonas. Analysis of housekeeping genes (rpoB, rpoD and gyrB) sequences showed similarities of 84-95% with respect to the closest related species of Pseudomonas, confirming its phylogenetic affiliation. The ANI values were less than 86% to the closest related species type strains. The respiratory quinone is Q9. The major fatty acids are C16:0, C16:1 ω7c/ C16:1 ω6c in summed feature 3 and C18:1 ω7c / C18:1 ω6c in summed feature 8. The strains are oxidase- and catalase-positive. Growth occurs at 4-30°C, and at pH 4.0-10. The DNA G+C content is 58.2-58.3mol %. The combined genotypic, phenotypic and chemotaxonomic data support the classification of strains L10.10T, A4R1.5 and A4R1.12 into a novel species of Pseudomonas, for which the name P. versuta sp. nov. is proposed. The type strain is L10.10T (LMG 29628T, DSM 101070T).
  13. Fulltext Characterization of a novel N-acylhomoserine lactonase, AidP, from Antarctic Planococcus sp
    See-Too WS, Convey P, Pearce DA, Chan KG
    Microb Cell Fact, 2018 Nov 17;17(1):179.
    PMID: 30445965 DOI: 10.1186/s12934-018-1024-6
    BACKGROUND: N-acylhomoserine lactones (AHLs) are well-studied signalling molecules produced by some Gram-negative Proteobacteria for bacterial cell-to-cell communication or quorum sensing. We have previously demonstrated the degradation of AHLs by an Antarctic bacterium, Planococcus versutus L10.15T, at low temperature through the production of an AHL lactonase. In this study, we cloned the AHL lactonase gene and characterized the purified novel enzyme.

    RESULTS: Rapid resolution liquid chromatography analysis indicated that purified AidP possesses high AHL-degrading activity on unsubstituted, and 3-oxo substituted homoserine lactones. Liquid chromatography-mass spectrometry analysis confirmed that AidP functions as an AHL lactonase that hydrolyzes the ester bond of the homoserine lactone ring of AHLs. Multiple sequence alignment analysis and phylogenetic analysis suggested that the aidP gene encodes a novel AHL lactonase enzyme. The amino acid composition analysis of aidP and the homologous genes suggested that it might be a cold-adapted enzyme, however, the optimum temperature is 28 °C, even though the thermal stability is low (reduced drastically above 32 °C). Branch-site analysis of several aidP genes of Planococcus sp. branch on the phylogenetic trees also showed evidence of episodic positive selection of the gene in cold environments. Furthermore, we demonstrated the effects of covalent and ionic bonding, showing that Zn2+ is important for activity of AidP in vivo. The pectinolytic inhibition assay confirmed that this enzyme attenuated the pathogenicity of the plant pathogen Pectobacterium carotovorum in Chinese cabbage.

    CONCLUSION: We demonstrated that AidP is effective in attenuating the pathogenicity of P. carotovorum, a plant pathogen that causes soft-rot disease. This anti-quorum sensing agent is an enzyme with low thermal stability that degrades the bacterial signalling molecules (AHLs) that are produced by many pathogens. Since the enzyme is most active below human body temperature (below 28 °C), and lose its activity drastically above 32 °C, the results of a pectinolytic inhibition assay using Chinese cabbage indicated the potential of this anti-quorum sensing agent to be safely applied in the field trials.

  14. Fulltext Complete genome of Arthrobacter alpinus strain R3.8, bioremediation potential unraveled with genomic analysis
    See-Too WS, Ee R, Lim YL, Convey P, Pearce DA, Mohidin TBM, et al.
    Stand Genomic Sci, 2017;12:52.
    PMID: 28904741 DOI: 10.1186/s40793-017-0264-0
    Arthrobacter alpinus R3.8 is a psychrotolerant bacterial strain isolated from a soil sample obtained at Rothera Point, Adelaide Island, close to the Antarctic Peninsula. Strain R3.8 was sequenced in order to help discover potential cold active enzymes with biotechnological applications. Genome analysis identified various cold adaptation genes including some coding for anti-freeze proteins and cold-shock proteins, genes involved in bioremediation of xenobiotic compounds including naphthalene, and genes with chitinolytic and N-acetylglucosamine utilization properties and also plant-growth-influencing properties. In this genome report, we present a complete genome sequence of A. alpinus strain R3.8 and its annotation data, which will facilitate exploitation of potential novel cold-active enzymes.
  15. Planococcus versutus sp. nov., isolated from soil
    See-Too WS, Ee R, Madhaiyan M, Kwon SW, Tan JY, Lim YL, et al.
    Int J Syst Evol Microbiol, 2017 Apr;67(4):944-950.
    PMID: 27959786 DOI: 10.1099/ijsem.0.001721
    A taxonomic study was performed on a novel Gram-stain-positive, coccus-shaped, orange-pigmented motile bacterium, designated as strain L10.15T. The organism was isolated from a soil sample collected in Lagoon Island (close to Adelaide Island, western Antarctic Peninsula) using a quorum-quenching enrichment medium. Growth occurred at 4-30 °C, pH 6-11 and at moderately high salinity (0-15 %, w/v, NaCl), with optimal growth at 26 °C, at pH 7-8 and with 6 % (w/v) NaCl. 16S rRNA gene sequence analysis showed that strain L10.15T belonged to the genus Planococcus and was closely related to Planococcus halocryophilus Or1T (99.3 % similarity), Planococcus donghaensis JH1T (99.0 %), Planococcus antarcticus DSM 14505T (98.3 %), Planococcus plakortidis AS/ASP6 (II)T (97.6 %), Planococcus maritimus TF-9T (97.5 %), Planococcus salinarum ISL-6T (97.5 %) and Planococcus kocurii NCIMB 629T (97.5 %). However, the average nucleotide identity-MUMmer analysis showed low genomic relatedness values of 71.1-81.7 % to the type strains of these closely related species of the genus Planococcus. The principal fatty acids were anteiso-C15 : 0, C16 : 1ω7c and anteiso-C17 :  0, and the major menaquinones of strain L10.15T were MK-5 (48 %), MK-6 (6 %) and MK-7 (44 %). Polar lipid analysis revealed the presence of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and aminophospholipid. The DNA G+C content was 39.4 mol%. The phenotypic and genotypic data indicate that strain L10.15T represents a novel species of the genus Planococcus, for which the name Planococcus versutus sp. nov. is proposed. The type strain is L10.15T (=DSM 101994T=KACC 18918T).
  16. Fulltext AidP, a novel N-Acyl homoserine lactonase gene from Antarctic Planococcus sp
    See-Too WS, Ee R, Lim YL, Convey P, Pearce DA, Yin WF, et al.
    Sci Rep, 2017 02 22;7:42968.
    PMID: 28225085 DOI: 10.1038/srep42968
    Planococcus is a Gram-positive halotolerant bacterial genus in the phylum Firmicutes, commonly found in various habitats in Antarctica. Quorum quenching (QQ) is the disruption of bacterial cell-to-cell communication (known as quorum sensing), which has previously been described in mesophilic bacteria. This study demonstrated the QQ activity of a psychrotolerant strain, Planococcus versutus strain L10.15T, isolated from a soil sample obtained near an elephant seal wallow in Antarctica. Whole genome analysis of this bacterial strain revealed the presence of an N-acyl homoserine lactonase, an enzyme that hydrolyzes the ester bond of the homoserine lactone of N-acyl homoserine lactone (AHLs). Heterologous gene expression in E. coli confirmed its functions for hydrolysis of AHLs, and the gene was designated as aidP (autoinducer degrading gene from Planococcus sp.). The low temperature activity of this enzyme suggested that it is a novel and uncharacterized class of AHL lactonase. This study is the first report on QQ activity of bacteria isolated from the polar regions.
  17. Fulltext Isolation, Characterisation, and Lipase Production of a Cold-Adapted Bacterial Strain Pseudomonas sp. LSK25 Isolated from Signy Island, Antarctica
    Salwoom L, Raja Abd Rahman RNZ, Salleh AB, Mohd Shariff F, Convey P, Pearce D, et al.
    Molecules, 2019 Feb 16;24(4).
    PMID: 30781467 DOI: 10.3390/molecules24040715
    In recent years, studies on psychrophilic lipases have been an emerging area of research in the field of enzymology. This study focuses on bacterial strains isolated from anthropogenically-influenced soil samples collected around Signy Island Research Station (South Orkney Islands, maritime Antarctic). Limited information on lipase activities from bacteria isolated from Signy station is currently available. The presence of lipase genes was determined using real time quantification PCR (qPCR) in samples obtained from three different locations on Signy Island. Twenty strains from the location with highest lipase gene detection were screened for lipolytic activities at a temperature of 4 °C, and from this one strain was selected for further examination based on the highest enzymatic activities obtained. Analysis of 16S rRNA sequence data of this strain showed the highest level of sequence similarity (98%) to a Pseudomonas sp. strain also isolated from Antarctica. In order to increase lipase production of this psychrophilic strain, optimisation of different parameters of physical and nutritional factors were investigated. Optimal production was obtained at 10 °C and pH 7.0, at 150 rev/min shaking rate over 36 h incubation.
  18. Fulltext New Recombinant Cold-Adapted and Organic Solvent Tolerant Lipase from Psychrophilic Pseudomonas sp. LSK25, Isolated from Signy Island Antarctica
    Salwoom L, Raja Abd Rahman RNZ, Salleh AB, Mohd Shariff F, Convey P, Mohamad Ali MS
    Int J Mol Sci, 2019 Mar 13;20(6).
    PMID: 30871178 DOI: 10.3390/ijms20061264
    In recent years, studies on psychrophilic lipases have become an emerging area of research in the field of enzymology. The study described here focuses on the cold-adapted organic solvent tolerant lipase strain Pseudomonas sp. LSK25 isolated from Signy Station, South Orkney Islands, maritime Antarctic. Strain LSK25 lipase was successfully cloned, sequenced, and over-expressed in an Escherichia coli system. Sequence analysis revealed that the lipase gene of Pseudomonas sp. LSK25 consists of 1432 bp, lacks an N-terminal signal peptide and encodes a mature protein consisting of 476 amino acids. The recombinant LSK25 lipase was purified by single-step purification using Ni-Sepharose affinity chromatography and had a molecular mass of approximately 65 kDa. The final recovery and purification fold were 44% and 1.3, respectively. The LSK25 lipase was optimally active at 30 °C and at pH 6. Stable lipolytic activity was reported between temperatures of 5⁻30 °C and at pH 6⁻8. A significant enhancement of lipolytic activity was observed in the presence of Ca2+ ions, the organic lipids of rice bran oil and coconut oil, a synthetic C12 ester and a wide range of water immiscible organic solvents. Overall, lipase strain LSK25 is a potentially desirable candidate for biotechnological application, due to its stability at low temperatures, across a range of pH and in organic solvents.
  19. Statistical optimisation of growth conditions and diesel degradation by the Antarctic bacterium, Rhodococcus sp. strain AQ5‒07
    Roslee AFA, Zakaria NN, Convey P, Zulkharnain A, Lee GLY, Gomez-Fuentes C, et al.
    Extremophiles, 2020 Mar;24(2):277-291.
    PMID: 31863235 DOI: 10.1007/s00792-019-01153-0
    Petroleum pollution is a major concern in Antarctica due to the persistent nature of its hydrocarbon components coupled with the region's extreme environmental conditions, which means that bioremediation approaches are largely inapplicable at present. The current study assessed the ability of the psychrotolerant phenol-degrader, Rhodococcus sp. strain AQ5-07, to assimilate diesel fuel as the sole carbon source. Factors expected to influence the efficiency of diesel degradation, including the initial hydrocarbon concentration, nitrogen source concentration and type, temperature, pH and salinity were studied. Strain AQ5-07 displayed optimal cell growth and biodegradation activity at 1% v/v initial diesel concentration, 1 g/L NH4Cl concentration, pH 7 and 1% NaCl during one-factor-at-a-time (OFAT) analyses. Strain AQ5-07 was psychrotolerant based on its optimum growth temperature being near 20 °C. In conventionally optimised media, strain AQ5-07 showed total petroleum hydrocarbons (TPH) mineralisation of 75.83%. However, the optimised condition for TPH mineralisation predicted through statistical response surface methodology (RSM) enhanced the reduction to 90.39% within a 2 days incubation. Our preliminary data support strain AQ5-07 being a potential candidate for real-field soil bioremediation by specifically adopting sludge-phase bioreactor system in chronically cold environments such as Antarctica. The study also confirmed the utility of RSM in medium optimisation.
  20. Fulltext Unveiling the Diversity of Periphytic Cyanobacteria (Cyanophyceae) from Tropical Mangroves in Penang, Malaysia
    Rahim NAA, Mohd Sidik Merican FM, Radzi R, Omar WMW, Nor SAM, Broady P, et al.
    Trop Life Sci Res, 2023 Sep;34(3):57-94.
    PMID: 37860087 DOI: 10.21315/tlsr2023.34.3.4
    Cyanobacteria are one of the most important groups of photoautotrophic organisms, contributing to carbon and nitrogen fixation in mangroves worldwide. They also play an important role in soil retention and stabilisation and contribute to high plant productivity through their secretion of plant growth-promoting substances. However, their diversity and distribution in Malaysian mangrove ecosystems have yet to be studied in detail, despite Malaysia hosting a significant element of remaining mangroves globally. In a floristic survey conducted in Penang, peninsular Malaysia, 33 morphospecies of periphytic cyanobacteria were identified and described for the first time from a mangrove ecosystem in Malaysia. Sixteen genera, comprising Aphanocapsa, Chroococcus, Chroococcidiopsis, Cyanobacterium, Desmonostoc, Geitlerinema, Leptolyngbya, Lyngbya, Microcystis, Myxosarcina, Oscillatoria, Phormidium, Pseudanabaena, Spirulina, Trichocoleus and Xenococcus, were obtained from field material growing on diverse natural and artificial substrata. Oscillatoriales was the dominant order with Phormidium the dominant genus at nine of the 15 sampling sites examined. Three of the morphospecies, Aphanocapsa cf. concharum, Xenococcus cf. pallidus and Oscillatoria pseudocurviceps, are rare and poorly known morphospecies worldwide. Chroococcus minutus, Phormidium uncinatum, P. amphigranulata, and some species of Oscillatoriales are considered as pollution indicator species. This study provides important baseline information for further investigation of the cyanobacterial microflora present in other mangrove areas around Malaysia. A complete checklist will enhance understanding of their ecological role and the potential for benefits arising from useful secondary metabolites or threats via toxin production to the ecosystem.
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