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An improved enzyme assay for molybdenum-reducing activity in bacteria

Shukor MY, Rahman MF, Shamaan NA, Lee CH, Karim MI, Syed MA

Appl Biochem Biotechnol, 2008 Mar;144(3):293-300.
PMID: 18556818

Molybdenum-reducing activity in the heterotrophic bacteria is a phenomenon that has been reported for more than 100 years. In the presence of molybdenum in the growth media, bacterial colonies turn to blue. The enzyme(s) responsible for the reduction of molybdenum to molybdenum blue in these bacteria has never been purified. In our quest to purify the molybdenum-reducing enzyme, we have devised a better substrate for the enzyme activity using laboratory-prepared phosphomolybdate instead of the commercial 12-phosphomolybdate we developed previously. Using laboratory-prepared phosphomolybdate, the highest activity is given by 10:4-phosphomolybdate. The apparent Michaelis constant, Km for the laboratory-prepared 10:4-phosphomolybdate is 2.56 +/- 0.25 mM (arbitrary concentration), whereas the apparent V(max) is 99.4 +/- 2.85 nmol Mo-blue min(-1) mg(-1) protein. The apparent Michaelis constant or Km for NADH as the electron donor is 1.38 +/- 0.09 mM, whereas the apparent V(max) is 102.6 +/- 1.73 nmol Mo-blue min(-1) mg(-l) protein. The apparent Km and V(max) for another electron donor, NADPH, is 1.43 +/- 0.10 mM and 57.16 +/- 1.01 nmol Mo-blue min(-1) mg(-1) protein, respectively, using the same batch of molybdenum-reducing enzyme. The apparent V(max) obtained for NADH and 10:4-phosphomolybdate is approximately 13 times better than 12-phoshomolybdate using the same batch of enzyme, and hence, the laboratory-prepared phosphomolybdate is a much better substrate than 12-phoshomolybdate. In addition, 10:4-phosphomolybdate can be routinely prepared from phosphate and molybdate, two common chemicals in the laboratory.

Matched MeSH terms: Enterobacter cloacae/enzymology*; Enterobacter cloacae/metabolism*
A new oxolane from Enterobacter cloacae

Yap AC, Chan KG, Sim KS, Choo YM

Nat Prod Res, 2016 Apr;30(7):783-8.
PMID: 26252083 DOI: 10.1080/14786419.2015.1065492

Enterobacter cloacae is a highly pathogenic Gram-negative proteobacterium which is responsible for a wide array of infections. In the present study, the fermentation culture of E. cloacae has yielded one new oxolane compound, Rimboxo (1) in addition to three known compounds, i.e. Maculosine (2), phenylacetic acid (3) and methyl myristate (4). These compounds were isolated and characterised using extensive chromatographic and spectroscopic methods, and were subjected to cytotoxicity evaluations.

Matched MeSH terms: Enterobacter cloacae
Fulltext Characterization of enterobacter cloacae isolated from street foods

Haryani, Y., Tunung, R., Chai, L.C., Lee, H.Y., Tang, S.Y., Son, R.

International Food Research Journal, 2008;15(1):57-64.
MyJurnal

A total of 78 samples comprising different types of street foods, sold in different locations in Malaysia, were examined for the presence of Enterobacter cloacae. E. cloacae contamination was recorded in 9% of the samples examined. Tests for susceptibility to 12 different antibiotics showed that all were resistant to six or more antibiotics, but susceptible to chloramphenicol and gentamicin. Plasmids of four different sizes were detected from the three plasmid positive isolates. RAPD analysis using four primers yielded completely different banding patterns for all E. cloacae studied. In Malaysia, no published information on street foods in the epidemiological investigation of E.cloacae related disease is available. However, their occurrences have provided compelling evidence that the risk of disease transmission caused by E. cloacae through street foods is moderate.

Matched MeSH terms: Enterobacter cloacae
Fulltext Larvicidal and anti-termite activities of microbial biosurfactant produced by Enterobacter cloacae SJ2 isolated from marine sponge Clathria sp

Harikrishnan S, Sudarshan S, Sivasubramani K, Nandini MS, Narenkumar J, Ramachandran V, et al.

Sci Rep, 2023 Sep 13;13(1):15153.
PMID: 37704703 DOI: 10.1038/s41598-023-42475-6

The widespread use of synthetic pesticides has resulted in a number of issues, including a rise in insecticide-resistant organisms, environmental degradation, and a hazard to human health. As a result, new microbial derived insecticides that are safe for human health and the environment are urgently needed. In this study, rhamnolipid biosurfactants produced from Enterobacter cloacae SJ2 was used to evaluate the toxicity towards mosquito larvae (Culex quinquefasciatus) and termites (Odontotermes obesus). Results showed dose dependent mortality rate was observed between the treatments. The 48 h LC50 (median lethal concentration) values of the biosurfactant were determined for termite and mosquito larvae following the non-linear regression curve fit method. Results showed larvicidal activity and anti-termite activity of biosurfactants with 48 h LC50 value (95% confidence interval) of 26.49 mg/L (25.40 to 27.57) and 33.43 mg/L (31.09 to 35.68), respectively. According to a histopathological investigation, the biosurfactant treatment caused substantial tissue damage in cellular organelles of larvae and termites. The findings of this study suggest that the microbial biosurfactant produced by E. cloacae SJ2 is an excellent and potentially effective agent for controlling Cx. quinquefasciatus and O. obesus.

Matched MeSH terms: Enterobacter cloacae
A new oxathiolane from Enterobacter cloacae

Yap AC, Teoh WY, Chan KG, Sim KS, Choo YM

Nat Prod Res, 2015;29(8):722-6.
PMID: 25427277 DOI: 10.1080/14786419.2014.983507

Enterobacter cloacae is a versatile bacterial species inhabiting a wide variety of niches and is capable of metabolising a wide variety of substances as energy resources. The fermentation culture of this bacterial species has successfully yielded one new compound, Rimboxa (1) and three known compounds, i.e. indole-3-carboxaldehyde (2), indole-3-acetic acid (3) and 3,4-di-t-butylaniline (4). Rimboxa (1) is shown to possess the 1,2-oxathiolane core structure. 3,4-Di-t-butylaniline (4) is isolated for the first time from a natural resource. These compounds were isolated and characterised using extensive chromatographic and spectroscopic methods, and were subjected to cytotoxicity evaluations.

Matched MeSH terms: Enterobacter cloacae/chemistry*
Fulltext Seed biopriming with P- and K-solubilizing Enterobacter hormaechei sp. improves the early vegetative growth and the P and K uptake of okra (Abelmoschus esculentus) seedling

Roslan MAM, Zulkifli NN, Sobri ZM, Zuan ATK, Cheak SC, Abdul Rahman NA

PLoS One, 2020;15(7):e0232860.
PMID: 32645001 DOI: 10.1371/journal.pone.0232860

Limited information is available that seed biopriming with plant growth-promoting Enterobacter spp. play a prominent role to enhance vegetative growth of plants. Contrary to Enterobacter cloacae, Enterobacter hormaechei is a less-studied counterpart despite its vast potential in plant growth-promotion mainly through the inorganic phosphorus (P) and potassium (K) solubilization abilities. To this end, 18 locally isolated bacterial pure cultures were screened and three strains showed high P- and K-solubilizing capabilities. Light microscopy, biochemical tests and 16S rRNA gene sequencing revealed that strains 15a1 and 40a were closely related to Enterobacter hormaechei while strain 38 was closely related to Enterobacter cloacae (Accession number: MN294583; MN294585; MN294584). All Enterobacter spp. shared common plant growth-promoting traits, namely nitrogen (N2) fixation, indole-3-acetic acid production and siderophore production. The strains 38 and 40a were able to produce gibberellic acid, while only strain 38 was able to secrete exopolysaccharide on agar. Under in vitro germination assay of okra (Abelmoschus esculentus) seeds, Enterobacter spp. significantly improved overall germination parameters and vigor index (19.6%) of seedlings. The efficacy of root colonization of Enterobacter spp. on the pre-treated seedling root tips was confirmed using Scanning Electron Microscopy (SEM). The pot experiment of bioprimed seeds of okra seedling showed significant improvement of the plant growth (> 28%) which corresponded to the increase of P and K uptakes (> 89%) as compared to the uninoculated control plants. The leaf surface area and the SPAD chlorophyll index of bioprimed plants were increased by up to 29% and 9% respectively. This report revealed that the under-explored species of P- and K-solubilizing Enterobacter hormaechei sp. with multiple plant beneficial traits presents a great potential sustainable approach for enhancement of soil fertility and P and K uptakes of plants.

Matched MeSH terms: Enterobacter cloacae
Isolation and characterization of a mo -reducing bacterium

Ghani B, Takai M, Hisham NZ, Kishimoto N, Ismail AK, Tano T, et al.

Appl Environ Microbiol, 1993 Apr;59(4):1176-80.
PMID: 16348915

A Mo -reducing bacterium (strain 48), which grew on medium supplemented with 200 mM Mo, was isolated from stream water obtained from Chengkau, Malaysia. The chemical properties of strain 48 conform to the characteristics of Enterobacter cloacae. Under anaerobic conditions in the glucose-yeast extract medium containing phosphate ion (2.9 mM) and Mo (10 mM), the bacterium reduced Mo to form molybdenum blue. Approximately 27% of Mo added to the medium was reduced after 28 h of cultivation. The reduction of Mo with glucose as an electron donor was strongly inhibited by iodoacetic acid, sodium fluoride, and sodium cyanide, suggesting an involvement of the glycolytic pathway and electron transport in Mo reduction. NADH and N,N,N',N' -tetramethyl-p-phenylenediamine served as electron donors for Mo reduction. When NADH was used as an electron donor, at first cytochrome b in the cell extract was reduced, and then molybdenum blue was formed. Sodium cyanide strongly inhibited Mo reduction by NADH (5 mM) but not the reduction of cytochrome b in the cell extract, suggesting that the reduced component of the electron transport system after cytochrome b serves as an electron donor for Mo reduction. Both ferric and stannous ions strongly enhanced the activity of Mo reduction by NADH.

Matched MeSH terms: Enterobacter cloacae
Role of coaggregation in the pathogenicity and prolonged colonisation of Vibrio cholerae

Toh YS, Yeoh SL, Yap IKS, Teh CSJ, Win TT, Thong KL, et al.

Med Microbiol Immunol, 2019 Dec;208(6):793-809.
PMID: 31263955 DOI: 10.1007/s00430-019-00628-3

Cholera is an acute diarrheal illness caused by the Gram-negative bacterium Vibrio cholerae. The pathogen is known for its ability to form biofilm that confers protection against harsh environmental condition and as part of the colonisation process during infection. Coaggregation is a process that facilitates the formation of biofilm. In a preliminary in vitro study, high coaggregation index and biofilm production were found between V. cholerae with human commensals namely Escherichia coli and Enterobacter cloacae. Building upon these results, the effects of coaggregation were further evaluated using adult BALB/c mouse model. The animal study showed no significant differences in mortality and fluid accumulation ratio between treatment groups infected with V. cholerae alone and those infected with coaggregation partnership (V. cholerae with E. coli or V. cholerae with E. cloacae). However, mild inflammation was detected in both partnering pairs. Higher density of V. cholerae was recovered from faecal samples of mice co-infected with E. coli and V. cholerae in comparison with other groups at 24 h post-infection. This partnership also elicited slightly higher levels of interleukin-5 (IL-5) and interleukin-10 (IL-10). Nonetheless, the involvement of autoinducer-2 (AI-2) as the signalling molecules in quorum sensing system is not evident in this study. Since E. coli is one of the common commensals, our result may suggest the involvement of commensals in cholera development.

Matched MeSH terms: Enterobacter cloacae/growth & development
Fulltext The effect of dietary bacterial organic selenium on growth performance, antioxidant capacity, and Selenoproteins gene expression in broiler chickens

Dalia AM, Loh TC, Sazili AQ, Jahromi MF, Samsudin AA

BMC Vet Res, 2017 Aug 18;13(1):254.
PMID: 28821244 DOI: 10.1186/s12917-017-1159-4

BACKGROUND: Selenium (Se) is an essential trace mineral in broilers, which has several important roles in biological processes. Organic forms of Se are more efficient than inorganic forms and can be produced biologically via Se microbial reduction. Hence, the possibility of using Se-enriched bacteria as feed supplement may provide an interesting source of organic Se, and benefit broiler antioxidant system and other biological processes. The objective of this study was to examine the impacts of inorganic Se and different bacterial organic Se sources on the performance, serum and tissues Se status, antioxidant capacity, and liver mRNA expression of selenoproteins in broilers.
RESULTS: Results indicated that different Se sources did not significantly (P ≤ 0.05) affect broiler growth performance. However, bacterial organic Se of T5 (basal diet +0.3 mg /kg feed ADS18 Se), T4 (basal diet +0.3 mg /kg feed ADS2 Se), and T3 (basal diet +0.3 mg /kg feed ADS1 Se) exhibited significantly (P ≤ 0.05) highest Se concentration in serum, liver, and kidney respectively. Dietary inorganic Se and bacterial organic Se were observed to significantly affect broiler serum ALT, AST, LDH activities and serum creatinine level. ADS18 supplemented Se of (Stenotrophomonas maltophilia) bacterial strain showed the highest GSH-Px activity with the lowest MDA content in serum, and the highest GSH-Px and catalase activity in the kidney, while bacterial Se of ADS2 (Klebsiella pneumoniae) resulted in a higher level of GSH-Px1 and catalase in liver. Moreover, our study showed that in comparison with sodium selenite, only ADS18 bacterial Se showed a significantly higher mRNA level in GSH-Px1, GSH-Px4, DIO1, and TXNDR1, while both ADS18 and ADS2 showed high level of mRNA of DIO2 compared to sodium selenite.
CONCLUSIONS: The supplementation of bacterial organic Se in broiler chicken, improved tissue Se deposition, antioxidant status, and selenoproteins gene expression, and can be considered as an effective alternative source of Se in broiler chickens.

Matched MeSH terms: Enterobacter cloacae/metabolism
Fulltext Antarctic bacteria inhibit the growth of pathogens

Wong, C.M.V.L., Chung, H.H., Aisyah, S., Omar, S., Cheah, Y.K., Maria de, L.G., et al.

ASM Science Journal, 2009;3(2):107-112.
MyJurnal

There are relatively little data on bacteria with antimicrobial activities from Antarctic, especially from the South Shetland Islands when compared to the other parts of the world. Hence, this project was set to isolate and characterize bacteria that produce anti-microbial compounds from Greenwich Island (one of the South Shetland Islands), Antarctica. A total of 356 strains of bacteria were isolated from Greenwich Island. They were screened for antimicrobial activities against 13 Gram-negative and one Gram-positive indicator food-borne pathogens. Two out of the 356 Antarctic bacterial strains exhibited an antagonistic effect on the indicator strains, Escherichia coli, Salmonella spp., Klebsiella pneumoniae, Enterobacter cloacae, Vibrio parahaemolyticus and Bacillus cereus. The two Antarctic bacterial strains were designated as SS157 and SR13. Biochemical and 16S rDNA analysis indicated that the strain SS157 was closely related to Pseudomonas congelans while the strain SR13 was closely related to Pseudomonas tremae. The anti-microbial compounds produced by the two Antarctic bacteria were not sensitive to temperature and were not degraded by trypsin or pronase indicating that they were likely to be chemical compounds or antibiotics. Antimicrobial compounds from strains SS157 and SR13 were broad spectrum, and targeted both Gram-positive and negative pathogens.

Matched MeSH terms: Enterobacter cloacae