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Extracellular Polyhydroxyalkanoate Depolymerase by Acidovorax sp. DP5.

Vigneswari S, Lee TS, Bhubalan K, Amirul AA - Enzyme Res (2015)

Bottom Line: The highest degradation activity of P(3HB) was also observed with depolymerase enzyme of DP5 in mineral salt medium containing P(3HB).Based on biochemical characterization and 16S rRNA cloning and sequencing, isolate DP5 was found to belong to the genus Acidovorax and subsequently named as Acidovorax sp.The highest extracellular depolymerase enzyme activity was achieved when 0.25% (w/v) of P(3HB) and 1 g/L of urea were used as carbon and nitrogen source, respectively, in the culture media.

View Article: PubMed Central - PubMed

Affiliation: Malaysian Institute of Pharmaceuticals and Nutraceuticals, NIBM, MOSTI, 11700 Penang, Malaysia ; Institute of Marine Biotechnology, Universiti Malaysia Terengganu, 21030 Kuala Terengganu, Malaysia.

ABSTRACT
Bacteria capable of degrading polyhydroxyalkanoates (PHA) by secreting extracellular depolymerase enzymes were isolated from water and soil samples collected from various environments in Malaysia. A total of 8 potential degraders exhibited clear zones on poly(3-hydroxybutyrate) [P(3HB)] based agar, indicating the presence of extracellular PHA depolymerase. Among the isolates, DP5 exhibited the largest clearing zone with a degradation index of 6.0. The highest degradation activity of P(3HB) was also observed with depolymerase enzyme of DP5 in mineral salt medium containing P(3HB). Based on biochemical characterization and 16S rRNA cloning and sequencing, isolate DP5 was found to belong to the genus Acidovorax and subsequently named as Acidovorax sp. DP5. The highest extracellular depolymerase enzyme activity was achieved when 0.25% (w/v) of P(3HB) and 1 g/L of urea were used as carbon and nitrogen source, respectively, in the culture media. The most suitable assay condition of the depolymerase enzyme in response to pH and temperature was tested. The depolymerase produced by strain Acidovorax sp. DP5 showed high percentage of degradation with P(3HB) films in an alkaline condition with pH 9 and at a temperature of 40°C.

No MeSH data available.


Related in: MedlinePlus

Neighbour-joining phylogenetic tree of Acidovorax sp. DP5 and related bacteria based on 16S rRNA sequence comparisons. Accession numbers are given.
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fig2: Neighbour-joining phylogenetic tree of Acidovorax sp. DP5 and related bacteria based on 16S rRNA sequence comparisons. Accession numbers are given.

Mentions: The result obtained (Table 1) showed that the microorganism coded as DP5 gave the highest degradation index of 6.00 followed by DP10 with the degradation index of 5.00. In order to determine the ability in producing P(3HB) degrading enzyme, enzyme assay of the 8 isolates was studied by using the samples at 48 h growth phase. Based on the enzyme activity assay, the DP5 bacterium showed the highest activity which was 0.035 mg/mL/min compared to the other isolates (Table 2). DP5 was found to exhibit the highest degradation index as well as depolymerase activity. DP5 was identified as Gram negative and rod shaped (Table 3). The 16S rRNA sequence of strain DP5 exhibited 99% similarity to Acidovorax delafieldii. Therefore, the strain was denoted as Acidovorax sp. DP5. The phylogenetic comparison of strain Acidovorax sp. DP5 within its genus is shown in Figure 2. In a separate study, Feng et al. reported the ability of Acidovorax sp. TP4 to degrade P(3HB) homopolymer and copolymer containing 3-hydroxyvalerate [30]. Salim et al. reported the presence of Acidovorax sp. and Ralstonia sp. from lake water and soil samples [24]. Based on Volovaa and coworkers, PHA-degrading Enterobacter sp., Bacillus sp., and Gracilibacillus sp. strains were isolated from tropical marine water [31].


Extracellular Polyhydroxyalkanoate Depolymerase by Acidovorax sp. DP5.

Vigneswari S, Lee TS, Bhubalan K, Amirul AA - Enzyme Res (2015)

Neighbour-joining phylogenetic tree of Acidovorax sp. DP5 and related bacteria based on 16S rRNA sequence comparisons. Accession numbers are given.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4664802&req=5

fig2: Neighbour-joining phylogenetic tree of Acidovorax sp. DP5 and related bacteria based on 16S rRNA sequence comparisons. Accession numbers are given.
Mentions: The result obtained (Table 1) showed that the microorganism coded as DP5 gave the highest degradation index of 6.00 followed by DP10 with the degradation index of 5.00. In order to determine the ability in producing P(3HB) degrading enzyme, enzyme assay of the 8 isolates was studied by using the samples at 48 h growth phase. Based on the enzyme activity assay, the DP5 bacterium showed the highest activity which was 0.035 mg/mL/min compared to the other isolates (Table 2). DP5 was found to exhibit the highest degradation index as well as depolymerase activity. DP5 was identified as Gram negative and rod shaped (Table 3). The 16S rRNA sequence of strain DP5 exhibited 99% similarity to Acidovorax delafieldii. Therefore, the strain was denoted as Acidovorax sp. DP5. The phylogenetic comparison of strain Acidovorax sp. DP5 within its genus is shown in Figure 2. In a separate study, Feng et al. reported the ability of Acidovorax sp. TP4 to degrade P(3HB) homopolymer and copolymer containing 3-hydroxyvalerate [30]. Salim et al. reported the presence of Acidovorax sp. and Ralstonia sp. from lake water and soil samples [24]. Based on Volovaa and coworkers, PHA-degrading Enterobacter sp., Bacillus sp., and Gracilibacillus sp. strains were isolated from tropical marine water [31].

Bottom Line: The highest degradation activity of P(3HB) was also observed with depolymerase enzyme of DP5 in mineral salt medium containing P(3HB).Based on biochemical characterization and 16S rRNA cloning and sequencing, isolate DP5 was found to belong to the genus Acidovorax and subsequently named as Acidovorax sp.The highest extracellular depolymerase enzyme activity was achieved when 0.25% (w/v) of P(3HB) and 1 g/L of urea were used as carbon and nitrogen source, respectively, in the culture media.

View Article: PubMed Central - PubMed

Affiliation: Malaysian Institute of Pharmaceuticals and Nutraceuticals, NIBM, MOSTI, 11700 Penang, Malaysia ; Institute of Marine Biotechnology, Universiti Malaysia Terengganu, 21030 Kuala Terengganu, Malaysia.

ABSTRACT
Bacteria capable of degrading polyhydroxyalkanoates (PHA) by secreting extracellular depolymerase enzymes were isolated from water and soil samples collected from various environments in Malaysia. A total of 8 potential degraders exhibited clear zones on poly(3-hydroxybutyrate) [P(3HB)] based agar, indicating the presence of extracellular PHA depolymerase. Among the isolates, DP5 exhibited the largest clearing zone with a degradation index of 6.0. The highest degradation activity of P(3HB) was also observed with depolymerase enzyme of DP5 in mineral salt medium containing P(3HB). Based on biochemical characterization and 16S rRNA cloning and sequencing, isolate DP5 was found to belong to the genus Acidovorax and subsequently named as Acidovorax sp. DP5. The highest extracellular depolymerase enzyme activity was achieved when 0.25% (w/v) of P(3HB) and 1 g/L of urea were used as carbon and nitrogen source, respectively, in the culture media. The most suitable assay condition of the depolymerase enzyme in response to pH and temperature was tested. The depolymerase produced by strain Acidovorax sp. DP5 showed high percentage of degradation with P(3HB) films in an alkaline condition with pH 9 and at a temperature of 40°C.

No MeSH data available.


Related in: MedlinePlus