Purification and characterization of an extracellular medium-chain length polyhydroxyalkanoate depolymerase from Thermus thermophilus HB8

Papaneophytou, Christos; Velali, Ekaterini; Pantazaki, Αnastasia A.

doi:10.1016/j.polymdegradstab.2010.12.015

Cited by 10 publications

(7 citation statements)

References 29 publications

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“…Moreover, the mcl PHA depolymerases described in this study contained a larger number of charged amino acids (ϳ18 to 22%) than the mature enzyme from P. fluorescens GK13 (15%). An increased number of intramolecular ion bonds by charged amino acids is known to contribute to the thermal stability of enzymes by conferring rigidity on them (36,43). This fact is in accordance with previous results that demonstrated that PhaZ SveSO1 showed higher thermostability than its PhaZ PflGK13 counterpart (8,47).…”

Section: Discussionsupporting

confidence: 92%

“…However, the ability to degrade scl PHA and mcl PHA by producing two types of depolymerases is rare and has been reported in only a few bacteria (5,23,25,36,47). In this work, the mcl PHA degraders S. roseolus SL3 and S. omiyaensis SO2 were also found to express scl PHA depolymerase in the presence of P(3HB).…”

Section: Discussionmentioning

confidence: 69%

“…However, only the PhaZ PflGK13 -coding gene and a few other homologous genes have been cloned and sequenced (21,29,37,50), including an mcl PHA depolymerase from the predator Bdellovibrio bacteriovorus (31). Additionally, in a recent work, a significant different gene from a thermophilic bacterium, T. thermophilus HB8, has been identified (36). However, no gene sequence of the genus Streptomyces has been reported so far.…”

Section: Discussionmentioning

confidence: 99%

“…In general, these enzymes consist of a signal peptide, an N-terminal substrate binding domain, and a C-terminal catalytic domain (15,22). In a recent study, the identification of a significantly different mcl PHA depolymerase gene from the thermophilic bacterium Thermus thermophilus HB8 has been reported (36). Recently, the isolation and identification of Streptomyces venezuelae SO1 as a novel mcl PHA depolymerase (PhaZ SveSO1 ) producer have been reported by our group (47).…”

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confidence: 92%

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Characterization of a Novel Subgroup of Extracellular Medium-Chain-Length Polyhydroxyalkanoate Depolymerases from Actinobacteria

Gangoiti

Santos

Prieto³

et al. 2012

Appl Environ Microbiol

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c Nineteen medium-chain-length (mcl) poly(3-hydroxyalkanoate) (PHA)-degrading microorganisms were isolated from natural sources. From them, seven Gram-positive and three Gram-negative bacteria were identified. The ability of these microorganisms to hydrolyze other biodegradable plastics, such as short-chain-length (scl) PHA, poly(-caprolactone) (PCL), poly(ethylene succinate) (PES), and poly(L-lactide) (PLA), has been studied. On the basis of the great ability to degrade different polyesters, Streptomyces roseolus SL3 was selected, and its extracellular depolymerase was biochemically characterized. The enzyme consisted of one polypeptide chain of 28 kDa with a pI value of 5.2. Its maximum activity was observed at pH 9.5 with chromogenic substrates. The purified enzyme hydrolyzed mcl PHA and PCL but not scl PHA, PES, and PLA. Moreover, the mcl PHA depolymerase can hydrolyze various substrates for esterases, such as tributyrin and p-nitrophenyl (pNP)-alkanoates, with its maximum activity being measured with pNP-octanoate. Interestingly, when poly(3-hydroxyoctanoate-co-3-hydroxyhexanoate [11%]) was used as the substrate, the main hydrolysis product was the monomer (R)-3-hydroxyoctanoate. In addition, the genes of several Actinobacteria strains, including S. roseolus SL3, were identified on the basis of the peptide de novo sequencing of the Streptomyces venezuelae SO1 mcl PHA depolymerase by tandem mass spectrometry. These enzymes did not show significant similarity to mcl PHA depolymerases characterized previously. Our results suggest that these distinct enzymes might represent a new subgroup of mcl PHA depolymerases.

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Section: Discussionsupporting

confidence: 92%

Section: Discussionmentioning

confidence: 69%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 92%

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Characterization of a Novel Subgroup of Extracellular Medium-Chain-Length Polyhydroxyalkanoate Depolymerases from Actinobacteria

Gangoiti

Santos

Prieto³

et al. 2012

Appl Environ Microbiol

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“…Therefore, there is increasing demand for developing novel methods that allow efficient production of RHAs, and the replacement of conventional processes by sustainable biotechnological procedures is one of the main current trends in white biotechnology and biocatalysis (Ren et al 2010). S. exfoliatus was found to be one of the few bacteria able to hydrolyze scl-and mcl-PHAs, indicating the presence of both types of depolymerases Elbanna et al 2004;Papaneophytou et al 2011;Martínez et al 2012;Santos et al 2013). Previously, scl-PHA depolymerase from S. exfoliatus DSMZ 41693 has been cloned and well characterized by our group .…”

Section: Discussionmentioning

confidence: 99%

Novel extracellular medium-chain-length polyhydroxyalkanoate depolymerase from Streptomyces exfoliatus K10 DSMZ 41693: a promising biocatalyst for the efficient degradation of natural and functionalized mcl-PHAs

Martínez

Santos

García-Hidalgo

et al. 2015

Appl Microbiol Biotechnol

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Cloning and biochemical characterization of a novel extracellular medium-chain-length polyhydroxyalkanoate (mcl-PHA) depolymerase from Streptomyces exfoliatus K10 DSMZ 41693 are described. The primary structure of the depolymerase (PhaZSex2) includes the lipase consensus sequence (serine-histidine-aspartic acid) which is known for serine hydrolases. Secondary structure analysis shows 7.9 % α-helix, 43.9 % β-sheet, 19.4 % β-turns, and 31.2 % random coil, suggesting that this enzyme belongs to the α/β hydrolase fold family, in agreement with other PHA depolymerases and lipases. The enzyme was efficiently produced as an extracellular active form in Rhodococcus and purified by two consecutive hydrophobic chromatographic steps. Matrix-assisted laser desorption-time-of-flight (MALDI-TOF) analysis of the purified enzyme revealed a monomer of 27.6 kDa with a midpoint transition temperature of 44.2 °C. Remarkably, the activity is significantly enhanced by low concentrations of nonionic and anionic detergents and thermal stability is improved by the presence of 10 % glycerol. PhaZSex2 is an endo-exohydrolase that cleaves both large and small PHA molecules, producing (R)-3-hydroxyoctanoic acid monomers as the main reaction product. Markedly, PhaZSex2 is able to degrade functionalized polymers containing thioester groups in the side chain (PHACOS), releasing functional thioester-based monomers and oligomers demonstrating the potentiality of this novel biocatalyst for the industrial production of enantiopure (R)-3-hydroxyalkanoic acids.

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Halevas¹,

Katsipis²,

Pantazaki³

2019

Biotechnological Applications of Polyhydroxyalkanoates

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Purification and characterization of an extracellular medium-chain length polyhydroxyalkanoate depolymerase from Thermus thermophilus HB8

Cited by 10 publications

References 29 publications

Characterization of a Novel Subgroup of Extracellular Medium-Chain-Length Polyhydroxyalkanoate Depolymerases from Actinobacteria

Characterization of a Novel Subgroup of Extracellular Medium-Chain-Length Polyhydroxyalkanoate Depolymerases from Actinobacteria

Novel extracellular medium-chain-length polyhydroxyalkanoate depolymerase from Streptomyces exfoliatus K10 DSMZ 41693: a promising biocatalyst for the efficient degradation of natural and functionalized mcl-PHAs

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