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1.
Substrate specificities of bacterial polyhydroxyalkanoate depolymerases and lipases: bacterial lipases hydrolyze poly(omega-hydroxyalkanoates). 总被引:3,自引:0,他引:3
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The substrate specificities of extracellular lipases purified from Bacillus subtilis, Pseudomonas aeruginosa, Pseudomonas alcaligenes, Pseudomonas fluorescens, and Burkholderia cepacia (former Pseudomonas cepacia) and of extracellular polyhydroxyalkanoate (PHA) depolymerases purified from Comamonas sp., Pseudomonas lemoignei, and P. fluorescens GK13, as well as that of an esterase purified from P. fluorescens GK 13, to various polyesters and to lipase substrates were analyzed. All lipases and the esterase of P. fluorescens GK13 but none of the PHA depolymerases tested hydrolyzed triolein, thereby confirming a functional difference between lipases and PHA depolymerases. However, most lipases were able to hydrolyze polyesters consisting of an omega-hydroxyalkanoic acid such as poly(6-hydroxyhedxanoate) or poly(4-hydroxybutyrate). The dimeric ester of hydroxyhexanoate was the main product of enzymatic hydrolysis of polycaprolactone by P. aeruginosa lipase. Polyesters containing side chains in the polymer backbone such as poly (3-hydroxybutyrate) and other poly(3-hydroxyalkanoates) were not or were only slightly hydrolyzed by the lipases tested. 相似文献
2.
Martínez V de la Peña F García-Hidalgo J de la Mata I García JL Prieto MA 《Applied and environmental microbiology》2012,78(17):6017-6026
The obligate predator Bdellovibrio bacteriovorus HD100 shows a large set of proteases and other hydrolases as part of its hydrolytic arsenal needed for its predatory life cycle. We present genetic and biochemical evidence that open reading frame (ORF) Bd3709 of B. bacteriovorus HD100 encodes a novel medium-chain-length polyhydroxyalkanoate (mcl-PHA) depolymerase (PhaZ(Bd)). The primary structure of PhaZ(Bd) suggests that this enzyme belongs to the α/β-hydrolase fold family and has a typical serine hydrolase catalytic triad (serine-histidine-aspartic acid) in agreement with other PHA depolymerases and lipases. PhaZ(Bd) has been extracellularly produced using different hypersecretor Tol-pal mutants of Escherichia coli and Pseudomonas putida as recombinant hosts. The recombinant PhaZ(Bd) has been characterized, and its biochemical properties have been compared to those of other PHA depolymerases. The enzyme behaves as a serine hydrolase that is inhibited by phenylmethylsulfonyl fluoride. It is also affected by the reducing agent dithiothreitol and nonionic detergents like Tween 80. PhaZ(Bd) is an endoexohydrolase that cleaves both large and small PHA molecules, producing mainly dimers but also monomers and trimers. The enzyme specifically degrades mcl-PHA and is inactive toward short-chain-length polyhydroxyalkanoates (scl-PHA) like polyhydroxybutyrate (PHB). These studies shed light on the potentiality of these predators as sources of new biocatalysts, such as an mcl-PHA depolymerase, for the production of enantiopure hydroxyalkanoic acids and oligomers as building blocks for the synthesis of biobased polymers. 相似文献
3.
Polyhydroxyalkanoates (PHAs) are a family of microbially made polyesters commercialized as biodegradable plastics. PHA production rates are predicted to increase as concerns around environmental plastic contamination and limited fossil fuel resources have increased the importance of biodegradable and bio-based plastic alternatives. Microbially produced PHA depolymerases are the key enzymes mediating PHA biodegradation, but only a few PHA depolymerases have been well-characterized and screens employing metagenomic sequence data are lacking. Here, we used 3078 metagenomes to analyse the distribution of PHA depolymerases in microbial communities from diverse aquatic, terrestrial and waste management systems. We significantly expand the recognized diversity of this protein family by screening 1914 Gb of sequence data and identifying 13 869 putative PHA depolymerases in 1295 metagenomes. Our results indicate that PHA depolymerases are unevenly distributed across environments. We predicted the highest frequency of PHA depolymerases in wastewater systems and the lowest in marine and thermal springs. In tandem, we screened 5290 metagenome-assembled genomes to describe the phylogenetic distribution of PHA depolymerases, which is substantially broader compared with current cultured representatives. The Proteobacteria and Bacteroidota are key lineages encoding PHA depolymerases, but PHA depolymerases were predicted from members of the Bdellovibrionota, Methylomirabilota, Actinobacteriota, Firmicutes, Spirochaetota, Desulfobacterota, Myxococcota and Planctomycetota. 相似文献
4.
Review Degradation of microbial polyesters 总被引:1,自引:0,他引:1
Microbial polyhydroxyalkanoates (PHAs), one of the largest groups of thermoplastic polyesters are receiving much attention as biodegradable substitutes for non-degradable plastics. Poly(D-3-hydroxybutyrate) (PHB) is the most ubiquitous and most intensively studied PHA. Microorganisms degrading these polyesters are widely distributed in various environments. Although various PHB-degrading microorganisms and PHB depolymerases have been studied and characterized, there are still many groups of microorganisms and enzymes with varying properties awaiting various applications. Distributions of PHB-degrading microorganisms, factors affecting the biodegradability of PHB, and microbial and enzymatic degradation of PHB are discussed in this review. We also propose an application of a new isolated, thermophilic PHB-degrading microorganism, Streptomyces strain MG, for producing pure monomers of PHA and useful chemicals, including D-3-hydroxycarboxylic acids such as D-3-hydroxybutyric acid, by enzymatic degradation of PHB. 相似文献
5.
《Biochimica et Biophysica Acta - Proteins and Proteomics》2020,1868(2):140315
Application of polyester-degrading enzymes should be considered as an eco-friendly alternative to chemical recycling due to the huge plastic waste disposal nowadays. Many hydrolases from several fungi and bacteria have been discovered and successfully evaluated for their activity towards different aliphatic polyesters (PHA, PBS, PBSA, PCL, PLA), aromatic polyesters (PET, PBT, PMT) as well as their co-polyesters (PBST, PBAT, PBSTIL). This revision gives an up-to-date overview on the main biochemical features and biotechnological applications of those reported enzymes which are able to degrade polyester-based plastics, including different microbial polyester depolymerases, esterases, cutinase-like enzymes and lipases. Summarized information includes available protein sequences with the corresponding accession numbers deposited in NCBI server, 3D resolved structures, and data about optimal conditions for enzymatic activity and stability of many of these microbial enzymes that would be helpful for researchers in this topic. Although screening and identification of new native polyester hydrolases from microbial sources is undeniable according to literature, we briefly highlight the importance of the design of improved enzymes towards recalcitrant aromatic polyesters through different approaches that include site-directed mutagenesis and surface protein engineering. 相似文献
6.
Biochemical and molecular characterization of the Pseudomonas lemoignei polyhydroxyalkanoate depolymerase system. 总被引:5,自引:2,他引:3
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D Jendrossek A Frisse A Behrends M Andermann H D Kratzin T Stanislawski H G Schlegel 《Journal of bacteriology》1995,177(3):596-607
Pseudomonas lemoignei has five different polyhydroxyalkanoate (PHA) depolymerase genes (phaZ1 to phaZ5), which encode the extracellularly localized poly(3-hydroxybutyrate) (PHB) depolymerases C, B, and D, poly(3-hydroxyvalerate) (PHV) depolymerase, and PHB depolymerase A, respectively. Four of the five genes (phaZ1 to phaZ4) have been cloned, and one of them (phaZ1) was studied in detail earlier (D. Jendrossek, B. Müller, and H. G. Schlegel, Eur. J. Biochem. 218:701-710, 1993). The fifth PHA depolymerase gene (phaZ5) was identified by colony hybridization of recombinant Escherichia coli clones with a phaZ5-specific oligonucleotide. The nucleotide sequence of a 3,704-bp EcoRI fragment was determined and found to contain two large open reading frames (ORFs) which coded for a polypeptide with significant similarities to glycerol-3-phosphate dehydrogenases of various sources (313 amino acids; M(r), 32,193) and for the precursor of PHB depolymerase A (PhaZ5; 433 amino acids; M(r), 44,906). The PHV depolymerase gene (phaZ4) was subcloned, and the nucleotide sequence of a 3,109-bp BamHI fragment was determined. Two large ORFs (ORF3 and ORF4) that represent putative coding regions were identified. The deduced amino acid sequence of ORF3 (134 amino acids; M(r), 14,686) revealed significant similarities to the branched-chain amino acid aminotransferase (IlfE) of enterobacteria. ORF4 (1,712 bp) was identified as the precursor of a PHV depolymerase (567 amino acids; M(r), 59,947). Analysis of primary structures of the five PHA depolymerases of P. lemoignei and of the PHB depolymerases of Alcaligenes faecalis and Pseudomonas pickettii revealed homologies of 25 to 83% to each other and a domain structure: at their N termini, they have typical signal peptides of exoenzymes. The adjacent catalytic domains are characterized by several conserved amino acids that constitute putative catalytic triads which consist of the consensus sequence of serine-dependent hydrolases including the pentapeptide G-X-S-X-G, a conserved histidine and aspartate, and a conserved region resembling the oxyanion hole of lipases. C terminal of the catalytic domain an approximately 40-amino-acid-long threonine-rich region (22 to 27 threonine residues) is present in PhaZ1, PhaZ2, PhaZ3, and PhaZ5. Instead of the threonine-rich region PhaZ4 and the PHB depolymerases of A. faecalis and P. pickettii contain an approximately 90-amino-acid-long sequence resembling the fibronectin type III module of eucaryotic extracellular matrix proteins. The function of the fibronectin type III module in PHA depolymerases remains obscure. Two types of C-terminal sequences apparently represent substrate-binding sites; the PHB type is present in the PHB depolymerases of A. faecalis and P. pickettii and in PhaZ2, PhaZ3, and PhaZ5 and the PHV type is present in the PHV-hydrolyzing depolymerases (PhaZ4 and PhaZ1). phaZ1 was transferred to A. eutrophus H16 and JMP222. All transconjugants of both strains were able to grow with extracellular PHB as a carbon source and produced translucent halos on PHB-containing solid media. PhaZ1, PhaZ2, PhaZ4, and PhaZ5 were purified from P. lemoignei and from recombinant E. coli; the processing sites of the precursors in E. coli were the same as in P. lemoignei, and similar substrate specificities were determined for the wild-type and the recombinant proteins. All PHA depolymerases hydrolyzed PHB at high specific activities. PhaZ1 and PhaZ4 additionally cleaved PHV, and PhaZ4 hydrolyzed poly(4-hydroxybutyrate). None of the depolymerases was able to hydrolyze polyactide or PHA consisting of monomers with more than five carbon atoms. While the wild-type depolymerase proteins were glycosylated and found to contain glucose and N-acetylglucosamine, none of the recombinant proteins was glycosylated. PHB hydrolysis was dependent on divalent cations such as Ca2+ and was inhibited by the presence of EDTA. 相似文献
7.
A Rhodospirillum rubrum gene that is predicted to code for an extracellular poly(3-hydroxybutyrate) (PHB) depolymerase by the recently published
polyhydroxyalkanoates (PHA) depolymerase engineering database was cloned. The gene product (PhaZ3
Rru
) was expressed in recombinant E. coli, purified and biochemically characterized. PhaZ3
Rru
turned out, however, to share characteristics of intracellular PHB depolymerases and revealed a combination of properties
that have not yet been described for other PHB depolymerases. A fusion of PhaZ3
Rru
with the enhanced cyan fluorescent protein was able to bind to PHB granules in vivo and supported the function as an intracellular
PHB depolymerase. Purified PhaZ3
Rru
was specific for short-chain-length polyhydroxyalkanoates (PHASCL) and hydrolysed both untreated native PHB granules as well as trypsin-activated native PHB granules to a mixture of mono-
and dimeric 3-hydroxybutyrate. Crystalline (denatured) PHB granules were not hydrolysed by PhayZ3
Rru
. Low concentrations of calcium or magnesium ions (1–5 mM) reversibly (EDTA) inhibited the enzyme. Our data suggest that PhaZ3
Rru
is the representative of a new type of the growing number of intracellular PHB depolymerases. 相似文献
8.
Biodegradation of polyhydroxyalkanoic acids 总被引:19,自引:0,他引:19
D. Jendrossek A. Schirmer H. G. Schlegel 《Applied microbiology and biotechnology》1996,46(5-6):451-463
Stimulated by the commercial availability of bacteriologically produced polyesters such as poly[(R)-3-hydroxybutyric acid], and encouraged by the discovery of new constituents of polyhydroxyalkanoic acids (PHA), a considerable
body of knowledge on the metabolism of PHA in microorganisms has accumulated. The objective of this essay is to give an overview
on the biodegradation of PHA. The following topics are discussed: (i) general considerations of PHA degradation, (ii) methods
for identification and isolation of PHA-degrading microorganisms, (iii) characterization of PHA-degrading microorganisms,
(iv) biochemical properties of PHA depolymerases, (v) mechanisms of PHA hydrolysis, (vi) regulation of PHA depolymerase synthesis,
(vii) molecular biology of PHA depolymerases, (viii) influence of the physicochemical properties of PHA on its biodegradability,
(ix) degradation of polyesters related to PHA, (x) biotechnological aspects of PHA and PHA depolymerases.
Received: 28 May 1996 / Received revision: 5 August 1996 / Accepted: 12 August 1996 相似文献
9.
A. N. Boyandin S. V. Prudnikova M. L. Filipenko E. A. Khrapov A. D. Vasil’ev T. G. Volova 《Applied Biochemistry and Microbiology》2012,48(1):28-36
Biodegradation of microbial linear polymers of hydroxyalkanoic acids (polyhydroxyalkanoates, PHAs) by soil microbial communities
of different structures has been studied during two field seasons in different weather conditions. This process was shown
to be influenced by the polymer chemical composition, temperature, humidity, and the microbial soil component. The PHA degradation
was accompanied by a decrease in the polymer molecular weight and an increase in the degree of crystallinity, indicating the
preferential destruction of the amorphous phase compared to the crystalline one. The quantity of the true PHA destructors
developing at the surface of the polymer samples was lower than the quantity of accompanying bacteria. The dominant PHA degrading
microorganisms under the test conditions were identified as bacteria of the genera Variovorax, Stenotrophomonas, Acinetobacter, Pseudomonas, Bacillus, and Xanthomonas and as micromycetes from Penicillium, Paecilomyces, Acremonium, Verticillium, and Zygosporium. 相似文献
10.
Elbanna K Lütke-Eversloh T Jendrossek D Luftmann H Steinbüchel A 《Archives of microbiology》2004,182(2-3):212-225
The biodegradability of microbial polythioesters (PTEs), a novel class of biopolymers which were discovered recently and can be produced by polyhydroxyalkanoate (PHA)-accumulating bacteria, was studied. Using poly(3-hydroxybutyrate-co-3-mercaptopropionate) [poly(3HB-co-3MP)] as sole carbon source for screening, 22 new bacterial strains were isolated and characterized. Interestingly, none of the PHA-degrading bacteria was able to utilize the homopolymer poly(3MP) as a carbon source for growth or to form clear zones on poly(3MP)-containing agar plates. The extracellular PHA depolymerases from two strains ( Schlegelella thermodepolymerans, Pseudomonas indica K2) were purified to electrophoretic homogeneity and biochemically characterized. The PHA depolymerase of S. thermodepolymerans exhibited a temperate optimum of about 75°C to 80°C and was stable at 70°C for more than 24 h. Regarding the substrate specificities of the PHA depolymerase of S. thermodepolymerans, enzyme activities decreased significantly with increasing 3MP content of the copolymer substrates. Interestingly, no activity could be detected with homoPTEs consisting only of 3MP or of 3-mercaptobutyrate. Similar results were obtained with the PHA depolymerases PhaZ2, PhaZ5 and PhaZ7 of Paucimonas lemoignei which were also investigated. The PHA depolymerase of Ps. indica K2 did not cleave any of the investigated polymers containing 3MP. Gas chromatography, infrared and 1H-NMR spectrometry and matrix-assisted laser desorption/ionization time-of-flight analysis revealed that 3MPs containing oligomers were enriched in the water-insoluble fraction remaining after partial digestion of poly(3HB-co-3MP) by purified poly(3HB) depolymerase of S. thermodepolymerans. In contrast, 3HB was enriched in the water-soluble fraction, which also contained 3HB-co-3MP dimer obtained by partial digestion of this copolymer by the enzyme. This study clearly indicates that PHA depolymerases are specific for oxoester linkages of PHAs and that the thioester bonds of PTEs cannot be cleaved by this type of enzyme.This publication is dedicated to Prof. Dr. Hans G. Schlegel in honor of his 80th birthday 相似文献
11.
Microbial polyhydroxyalkanoates (PHA) are a family of structurally diverse polyesters produced by many bacteria. Deleting key steps from the beta-oxidation cycle in Pseudomonas putida makes it possible to achieve precise substrate based design of PHA homopolymers, copolymers, and block polymers, allowing the study of structure-property relationship in a clear way. The PHA homopolymer synthesis also allows the microbial or chemical production of pure monomers of PHA in a convenient way without separating the mixed monomers. After used as bioplastics, PHA can be methyl esterified to become biofuels, which further extends the PHA application value. The microbial production of PHA with diverse structures is entering a new developing phase. 相似文献
12.
Boiandin AN Prudnikova SV Filipenko ML Khrapov EA Vasil'ev AD Volova TG 《Prikladnaia biokhimiia i mikrobiologiia》2012,48(1):35-44
Biodegradation of microbial linear polymers of hydroxyalkanoic acids (polyhydroxyalkanoates, PHAs) by soil microbiocoenoses of different structures has been studied during two field seasons in different weather conditions. This process was shown to be influenced by the polymer chemical composition, temperature, humidity, and the microbial soil component. The PHA degradation was accompanied by a decrease in the polymer molecular weight and an increase in the degree of crystallinity, indicating the preferential destruction of the amorphous phase compared to the crystalline one. The quantity of the true PHA destructors developing at the surface of the polymer samples was lower than the quantity of accompanying bacteria. The dominant PHA destructors under the test conditions were identified as bacteria of the genera Variovorax, Stenotrophomonas, Acinetobacter, Pseudomonas, Bacillus, and Xanthomonas and as micromycetes from Penicillium, Paecilomyces, Acremonium, Verticillium. and Zygosporium. 相似文献
13.
利用单一微生物发酵是现阶段获得聚羟基脂肪酸酯 (PHA) 的主要方式,但过高的生产成本限制了其大规模应用。近年来利用活性污泥菌群混合培养合成PHA被广泛研究。将剩余污泥处理与PHA合成相结合,不仅可以省去纯培养所必需的灭菌环节,同时可以实现剩余污泥的资源化利用。剩余污泥的水解酸化、菌群富集驯化及PHA合成受环境因素影响,深入的生物合成机制研究有助于混合培养合成PHA的推广应用。文中主要介绍利用剩余污泥合成PHA的可行性、影响剩余污泥水解酸化的因素、污泥菌群富集驯化合成PHA及其机制等方面的研究进展。 相似文献
14.
Background
PhaR which is a repressor protein for microbial polyhydroxyalkanoates (PHA) biosynthesis, is able to attach to bacterial PHA granules in vivo, was developed as an affinity tag for in vitro protein purification. Fusion of PhaR-tagged self-cleavable Ssp DnaB intein to the N-terminus of a target protein allowed protein purification with a pH and temperature shift. During the process, the target protein was released to the supernatant while PhaR-tagged intein was still immobilized on the PHA nanoparticles which were then separated by centrifugation. 相似文献15.
Microbial degradation of polyurethane, polyester polyurethanes and polyether polyurethanes 总被引:9,自引:0,他引:9
Nakajima-Kambe T Shigeno-Akutsu Y Nomura N Onuma F Nakahara T 《Applied microbiology and biotechnology》1999,51(2):134-140
Polyurethane (PUR) is a polymer derived from the condensation of polyisocyanate and polyol and it is widely used as a base
material in various industries. PUR, in particular, polyester PUR, is known to be vulnerable to microbial attack. Recently,
environmental pollution by plastic wastes has become a serious issue and polyester PUR had attracted attention because of
its biodegradability. There are many reports on the degradation of polyester PUR by microorganisms, especially by fungi. Microbial
degradation of polyester PUR is thought to be mainly due to the hydrolysis of ester bonds by esterases. Recently, polyester-PUR-degrading
enzymes have been purified and their characteristics reported. Among them, a solid-polyester-PUR-degrading enzyme (PUR esterase)
derived from Comamonas acidovorans TB-35 had unique characteristics. This enzyme has a hydrophobic PUR-surface-binding domain and a catalytic domain, and the
surface-binding domain was considered as being essential for PUR degradation. This hydrophobic surface-binding domain is also
observed in other solid-polyester-degrading enzymes such as poly(hydroxyalkanoate) (PHA) depolymerases. There was no significant
homology between the amino acid sequence of PUR esterase and that of PHA depolymerases, except in the hydrophobic surface-binding
region. Thus, PUR esterase and PHA depolymerase are probably different in terms of their evolutionary origin and it is possible
that PUR esterases come to be classified as a new solid-polyester-degrading enzyme family.
Received: 20 July 1998 / Received revision: 9 October 1998 / Accepted: 11 October 1998 相似文献
16.
The integrated study addressing biodegradation of microbial linear polyesters of hydroxyalkanoic acids (polyhydroxyalkanoates, PHAs) in tropical conditions by microbial communities of Vietnamese soils was performed in locations close to Hanoi and Nha Trang, which differed in their weather conditions and microbial communities. It shows that PHA degradation in tropical soils is influenced by polymer chemical composition, specimen shape, and microbial characteristics. The homopolymer of 3-hydroxybutyric acid is degraded at higher rates than the copolymer of 3-hydroxybutyric and 3-hydroxyvaleric acids. The average rates of mass loss were 0.04–0.33% per day for films and 0.02–0.18% for compact pellets. PHA degradation was accompanied by a decrease in the polymer molecular mass and, usually, an increase in the degree of crystallinity, suggesting preferential degradation of the amorphous phase. Under the study conditions, representatives of the bacterial genera Burkholderia, Bacillus, Cupriavidus, Mycobacterium, and Nocardiopsis and such micromycetes as Acremonium, Gongronella, Paecilomyces, and Penicillium, Trichoderma have been identified as major PHA degraders. 相似文献
17.
PHA synthase is the key enzyme involved in the biosynthesis of microbial polymers, polyhydroxyalkanoates (PHA). In this study, we created a hybrid library of PHA synthase gene with different crossover points by an incremental truncation method between the C-terminal fragments of the phaC(Cn) (phaC from Cupriavidus necator) and the N-terminal fragments of the phaC1(Pa) (phaC from Pseudomonas aeruginosa). As the truncation of the hybrid enzyme increased, the in vivo PHB synthesis ability of the hybrids declined gradually. PHA synthase PhaC(Cn) with a deletion on N-terminal up to 83 amino acid residues showed no synthase activity. While with the removal of up to 270 amino acids from the N-terminus, the activity of the truncated PhaC(Cn) could be complemented by the N-terminus of PhaC1(Pa). Three of the hybrid enzymes W188, W235 and W272 (named by the deleted nucleic acid number) were found to have altered product specificities. 相似文献
18.
Michael Knoll Thomas M Hamm Florian Wagner Virginia Martinez Jürgen Pleiss 《BMC bioinformatics》2009,10(1):89
Background
Polyhydroxyalkanoates (PHAs) can be degraded by many microorganisms using intra- or extracellular PHA depolymerases. PHA depolymerases are very diverse in sequence and substrate specificity, but share a common α/β-hydrolase fold and a catalytic triad, which is also found in other α/β-hydrolases. 相似文献19.
Moralejo-Gárate H Mar'atusalihat E Kleerebezem R van Loosdrecht MC 《Applied microbiology and biotechnology》2011,92(3):631-639
In this work, the potential of using microbial community engineering for production of polyhydroxyalkanoates (PHA) from glycerol
was explored. Crude glycerol is a by-product of the biofuel (biodiesel and bioethanol) industry and potentially a good substrate
for bioplastic production. A PHA-producing microbial community was enriched based on cultivation in a feast–famine regime
as successfully applied before for fatty acids-based biopolymer production. A glycerol-fed sequencing batch reactor operated
at a 2-day liquid and biomass residence time and with feast–famine cycles of 24 h was used to enrich a mixed community of
PHA producers. In a subsequent fed-batch PHA production step under growth-limiting conditions, the enriched mixed community
produced PHA up to a dry weight content of 80 wt.%. The conversion efficiency of substrate to PHA on electron basis was 53%.
Since glycerol is entering the metabolic pathways of the cell in the glycolytic pathway, it was anticipated that besides PHA,
polyglucose could be formed as storage polymer as well. Indeed, polyglucose was produced in low amounts (∼10 wt.%). The results
indicated that the feast–famine-based enrichment strategy was comparably successful to obtain a microbial community compared
to fatty acids-based enrichment described before. 相似文献
20.
Joana Gangoiti Marta Santos María Auxiliadora Prieto Isabel de la Mata Juan L. Serra María J. Llama 《Applied and environmental microbiology》2012,78(20):7229-7237
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(ε-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. 相似文献