Colonization and degradation of polyhydroxyalkanoates by lipase-producing bacteria

Sharma, Parveen; Mohanan, Nisha; Sidhu, Ravinder; Levin, David B.

doi:10.1139/cjm-2019-0042

Cited by 13 publications

(9 citation statements)

References 51 publications

(48 reference statements)

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“…Of these, three encode for intracellular lipases (EY04_08410, EY04_09635, EY04_02420), three encode a signal peptide and are extracellular lipases (EY04_21540, EY04_17885, EY04_32435), and one encodes for an intracellular PHA depolymerase (EY04_01535). In our previous study, the hydrolysis of the ester bonds of p-nitrophenyl-fatty acid substrates, and the ester bonds of PHA polymers of various subunit composition by the extracellular esterases/lipases was recorded in the culture media in which P. chlororaphis PA23 and A. lwoffii were grown ( Sharma et al, 2019 ). In the present study, two genes encoding intracellular lipases were cloned and expressed in E. coli BL21(DE3) to compare the properties and investigate its biodegradability in comparison with various biodegradable polymers.…”

Section: Resultsmentioning

confidence: 99%

“…In a previous report from this lab( Sharma et al, 2019 ), the cell free culture supernatant of Pseudomonas chlororaphis PA23 was shown to hydrolyze the ester bonds of p-nitrophenyl fatty acid substrates and PHA polymers with different subunit composition. In the present study, two genes encoding intracellular lipases from P. chlororaphis PA23 were cloned and expressed in E. coli BL21(DE3) to study the enzyme properties and investigate their degradation activity toward various biodegradable polymers such as small chain length (scl-) and mcl-PHAs, polylactic acid (PLA), polyethylene succinate (PES), and PCL.…”

Section: Introductionmentioning

confidence: 95%

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Characterization of Polymer Degrading Lipases, LIP1 and LIP2 From Pseudomonas chlororaphis PA23

Mohanan

Wong

Budiša

et al. 2022

Front. Bioeng. Biotechnol.

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The outstanding metabolic and bioprotective properties of the bacterial genus Pseudomonas make these species a potentially interesting source for the search of hydrolytic activities that could be useful for the degradation of plastics. We identified two genes encoding the intracellular lipases LIP1 and LIP2 of the biocontrol bacterium Pseudomonas chlororaphis PA23 and subsequently performed cloning and expression in Escherichia coli. The lip1 gene has an open reading frame of 828 bp and encodes a protein of 29.7 kDa whereas the lip2 consists of 834 bp and has a protein of 30.2 kDa. Although secondary structure analyses of LIP1 and LIP2 indicate a dominant α/β-hydrolase-fold, the two proteins differ widely in their amino acid sequences (15.39% identity), substrate specificities, and hydrolysis rates. Homology modeling indicates the catalytic serine in both enzymes located in a GXSXG sequence motif (lipase box). However, LIP1 has a catalytic triad of Ser152-His253-Glu221 with a GGX-type oxyanion pocket, whereas LIP2 has Ser138-His249-Asp221 in its active site and a GX-type of oxyanion hole residues. However, LIP1 has a catalytic triad of Ser152-His253-Glu221 with an oxyanion pocket of GGX-type, whereas LIP2 has Ser138-His249-Asp221 in its active site and a GX-type of oxyanion hole residues. Our three-dimensional models of LIP1 and LIP2 complexed with a 3-hydroxyoctanoate dimer revealed the core α/β hydrolase-type domain with an exposed substrate binding pocket in LIP1 and an active-site capped with a closing lid domain in LIP2. The recombinant LIP1 was optimally active at 45°C and pH 9.0, and the activity improved in the presence of Ca2+. LIP2 exhibited maximum activity at 40°C and pH 8.0, and was unaffected by Ca2+. Despite different properties, the enzymes exhibited broadsubstrate specificity and were able to hydrolyze short chain length and medium chain length polyhydroxyalkanoates (PHAs), polylactic acid (PLA), and para-nitrophenyl (pNP) alkanoates. Gel Permeation Chromatography (GPC) analysis showed a decrease in the molecular weight of the polymers after incubation with LIP1 and LIP2. The enzymes also manifested some polymer-degrading activity on petroleum-based polymers such as poly(ε-caprolactone) (PCL) and polyethylene succinate (PES), suggesting that these enzymes could be useful for biodegradation of synthetic polyester plastics. The study will be the first report of the complete characterization of intracellular lipases from bacterial and/or Pseudomonas species. The lipases, LIP1 and LIP2 are different from other bacterial lipases/esterases in having broad substrate specificity for polyesters.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 95%

Characterization of Polymer Degrading Lipases, LIP1 and LIP2 From Pseudomonas chlororaphis PA23

Mohanan

Wong

Budiša

et al. 2022

Front. Bioeng. Biotechnol.

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“…Nitrosomonas have been proposed as a PHA-producing bacterium (Yang et al 2013). Previous reports have suggested that many bacteria, including denitrifiers, produce lipases rather than polymerases to degrade PHAs, though the reason is not known (Chu and Wang 2017, Jaeger et al 1995, Muhammadi et al 2015, Sharma et al 2019, Wang and Chu 2016. We hypothesized that either anaerobic condition or the presence of PHA or other bacteria might induce the lipase expression.…”

Section: Which Are the Lipolytic Mags And Why They Have Lipases?mentioning

confidence: 92%

Looking for lipases and lipolytic organisms in low-temperature anaerobic reactors treating domestic wastewater

Bashiri

Allen

Shamurad

et al. 2021

Preprint

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Poor lipid degradation limits low-temperature anaerobic treatment of domestic wastewater even when psychrophiles are used. We combined metagenomics and metaproteomics to find lipolytic bacteria and their potential, and actual, cold-adapted extracellular lipases in anaerobic membrane bioreactors treating domestic wastewater at 4°C and 15°C. Of the 40 recovered putative lipolytic metagenome-assembled genomes (MAGs), only three (Chlorobium, Desulfobacter, and Mycolicibacterium) were common and abundant (relative abundance ≥ 1%) in all reactors. Notably, some MAGs that represented aerobic autotrophs contained lipases. Therefore, we hypothesised that the lipases we found are not always associated with exogenous lipid degradation and can have other roles such as polyhydroxyalkanoates (PHA) accumulation/degradation and interference with the outer membranes of other bacteria. Metaproteomics did not provide sufficient proteome coverage for relatively lower abundant proteins such as lipases though the expression of fadL genes, long-chain fatty acid transporters, was confirmed for four genera (Dechloromonas, Azoarcus, Aeromonas and Sulfurimonas), none of which were recovered as putative lipolytic MAGs. Metaproteomics also confirmed the presence of 15 relatively abundant (≥1%) genera in all reactors, of which at least 6 can potentially accumulate lipid/polyhydroxyalkanoates. For most putative lipolytic MAGs, there was no statistically significant correlation between the read abundance and reactor conditions such as temperature, phase (biofilm and bulk liquid), and feed type (treated by ultraviolet light or not). Results obtained by metagenomics and metaproteomics did not confirm each other and further work is required to identify the true lipid degraders in these systems.

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“…Nitrosomonas have been proposed as a PHA-producing bacterium (Yang et al, 2013). Previous reports have suggested that many bacteria, including denitrifiers, produce lipases rather than polymerases to degrade PHAs, though the reason is not known (Chu and Wang, 2017;Jaeger et al, 1995;Muhammadi et al, 2015;Sharma et al, 2019;Wang and Chu, 2016). We hypothesized that either anaerobic condition or the presence of PHA or other bacteria might induce the lipase expression.…”

Section: Which Are the Lipolytic Mags And Why They Have Lipases?mentioning

confidence: 95%

Looking for lipases and lipolytic organisms in low-temperature anaerobic reactors treating domestic wastewater

et al. 2022

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Colonization and degradation of polyhydroxyalkanoates by lipase-producing bacteria

Cited by 13 publications

References 51 publications

Characterization of Polymer Degrading Lipases, LIP1 and LIP2 From Pseudomonas chlororaphis PA23

Characterization of Polymer Degrading Lipases, LIP1 and LIP2 From Pseudomonas chlororaphis PA23

Looking for lipases and lipolytic organisms in low-temperature anaerobic reactors treating domestic wastewater

Looking for lipases and lipolytic organisms in low-temperature anaerobic reactors treating domestic wastewater

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