Synthesis of Short-Chain-Length and Medium-Chain-Length Polyhydroxyalkanoate Blends from Activated Sludge by Manipulating Octanoic Acid and Nonanoic Acid as Carbon Sources

2018 ◽  
Vol 66 (42) ◽  
pp. 11043-11054 ◽  
Author(s):  
Zheng Chen ◽  
Chuanpan Zhang ◽  
Liang Shen ◽  
Heng Li ◽  
Yajuan Peng ◽  
...  
Keyword(s):  
Activated Sludge ◽  
Chain Length ◽  
Octanoic Acid ◽  
Carbon Sources ◽  
Short Chain ◽  
Nonanoic Acid ◽  
Short Chain Length ◽  
Medium Chain ◽  
Medium Chain Length

scholarly journals Characterization of Site-Specific Mutations in a Short-Chain-Length/Medium-Chain-Length Polyhydroxyalkanoate Synthase:In VivoandIn VitroStudies of Enzymatic Activity and Substrate Specificity

2013 ◽  
Vol 79 (12) ◽  
pp. 3813-3821 ◽  
Author(s):  
Jo-Ann Chuah ◽  
Satoshi Tomizawa ◽  
Miwa Yamada ◽  
Takeharu Tsuge ◽  
Yoshiharu Doi ◽  
...  
Keyword(s):  
Chain Length ◽  
Fold Increase ◽  
Short Chain ◽  
Pha Synthase ◽  
Wild Type ◽  
Short Chain Length ◽  
Medium Chain ◽  
Medium Chain Length

ABSTRACTSaturation point mutagenesis was carried out at position 479 in the polyhydroxyalkanoate (PHA) synthase fromChromobacteriumsp. strain USM2 (PhaCCs) with specificities for short-chain-length (SCL) [(R)-3-hydroxybutyrate (3HB) and (R)-3-hydroxyvalerate (3HV)] and medium-chain-length (MCL) [(R)-3-hydroxyhexanoate (3HHx)] monomers in an effort to enhance the specificity of the enzyme for 3HHx. A maximum 4-fold increase in 3HHx incorporation and a 1.6-fold increase in PHA biosynthesis, more than the wild-type synthase, was achieved using selected mutant synthases. These increases were subsequently correlated with improved synthase activity and increased preference of PhaCCsfor 3HHx monomers. We found that substitutions with uncharged residues were beneficial, as they resulted in enhanced PHA production and/or 3HHx incorporation. Further analysis led to postulations that the size and geometry of the substrate-binding pocket are determinants of PHA accumulation, 3HHx fraction, and chain length specificity.In vitroactivities for polymerization of 3HV and 3HHx monomers were consistent within vivosubstrate specificities. Ultimately, the preference shown by wild-type and mutant synthases for either SCL (C4and C5) or MCL (C6) substrates substantiates the fundamental classification of PHA synthases.

scholarly journals Coexpression of Genetically Engineered 3-Ketoacyl-ACP Synthase III (fabH) and Polyhydroxyalkanoate Synthase (phaC) Genes Leads to Short-Chain-Length-Medium-Chain-Length Polyhydroxyalkanoate Copolymer Production from Glucose in Escherichia coli JM109

2004 ◽  
Vol 70 (2) ◽  
pp. 999-1007 ◽  
Author(s):  
Christopher T. Nomura ◽  
Kazunori Taguchi ◽  
Seiichi Taguchi ◽  
Yoshiharu Doi
Keyword(s):  
Escherichia Coli ◽  
Chain Length ◽  
Acyl Carrier Protein ◽  
Short Chain ◽  
Pha Synthase ◽  
Short Chain Length ◽  
E Coli ◽  
Medium Chain ◽  
Medium Chain Length ◽  
Pha Copolymers

ABSTRACT Polyhydroxyalkanoates (PHAs) can be divided into three main types based on the sizes of the monomers incorporated into the polymer. Short-chain-length (SCL) PHAs consist of monomer units of C3 to C5, medium-chain-length (MCL) PHAs consist of monomer units of C6 to C14, and SCL-MCL PHAs consist of monomers ranging in size from C4 to C14. Although previous studies using recombinant Escherichia coli have shown that either SCL or MCL PHA polymers could be produced from glucose, this study presents the first evidence that an SCL-MCL PHA copolymer can be made from glucose in recombinant E. coli. The 3-ketoacyl-acyl carrier protein synthase III gene (fabH) from E. coli was modified by saturation point mutagenesis at the codon encoding amino acid 87 of the FabH protein sequence, and the resulting plasmids were cotransformed with either the pAPAC plasmid, which harbors the Aeromonas caviae PHA synthase gene (phaC), or the pPPAC plasmid, which harbors the Pseudomonas sp. strain 61-3 PHA synthase gene (phaC1), and the abilities of these strains to accumulate PHA from glucose were assessed. It was found that overexpression of several of the mutant fabH genes enabled recombinant E. coli to induce the production of monomers of C4 to C10 and subsequently to produce unusual PHA copolymers containing SCL and MCL units. The results indicate that the composition of PHA copolymers may be controlled by the monomer-supplying enzyme and further reinforce the idea that fatty acid biosynthesis may be used to supply monomers for PHA production.

The Production and Analysis of Biodegradable Polymers of Type of Medium-Chain-Length Polyhydroxyalkanoates (mcl-PHA) by Pseudomonas putida Strain for the Biomedical Engineering

2021 ◽  
Vol 22 ◽  
Author(s):  
Nicoleta Ene ◽  
Mariana-Gratiela Vladu ◽  
Irina Lupescu ◽  
Ana-Despina Ionescu ◽  
Emanuel Vamanu
Keyword(s):  
Pseudomonas Putida ◽  
Chain Length ◽  
Octanoic Acid ◽  
Dry Weight ◽  
Growth Conditions ◽  
Spectrophotometric Analysis ◽  
Nonanoic Acid ◽  
Mechanical Resistance ◽  
Medium Chain ◽  
Medium Chain Length

Background: Polyhydroxyalkanoates (PHAs) are bacteria-synthetized biopolymers under unbalanced growth conditions. These biopolymers are considered potential biomaterials for future applications for their biocompatibility and biodegradable features and potential biomaterials for future applications for their biocompatibility and biodegradable characteristics and their ability to be quickly produced and functionalize with strong mechanical resistance. This article is intended to perform microbial fermentation using Pseudomonas putida strain to show the amount of biopolymers of the type polyhydroxyalkanoates with medium-chain-length (mcl-PHA) obtained depending on the type and quantity of added precursors (glucose and fatty acids). Methods: It is important to understand the microbial interaction and mechanism involved in PHA biosynthetis.For these, several methods were used, such as: obtaining microbial biomass by using a Pseudomonas putida strain able of PHA-producing, analysis of biopolymer production by acetone extraction following the Soxhlet method, purification of biopolymer by methanol-ethanol treatment, followed by the estimation of biomass by spectrophotometric analysis and the measurement of the dry weight of cells and the quantification of the amount of biopolymer produced following the gas chromatographic method (GC). Results: The highest PHA yield was obtained using octanoic (17 mL in 2000 mL medium) and hexanoic acids (14 mL in 2000 mL medium) as precursors. Consequently, octanoic acid – octanoic acid, heptanoic acid – nonanoic acid, and octanoic acid - hexanoic acid were the mix of precursors that supported the amount of PHA obtained. Conclusion: Of the 4 types of structurally related substrate, the strain Pseudomonas putida ICCF 319 prefers the C8 sublayer for an elastomeric PHA's biosynthesis with a composition in which the C8 monomer predominates over C6 and C10.

EngineeredEscherichia colifor Short-Chain-Length Medium-Chain-Length Polyhydroxyalkanoate Copolymer Biosynthesis from Glycerol and Dodecanoate

2013 ◽  
Vol 77 (6) ◽  
pp. 1262-1268 ◽  
Author(s):  
Chitwadee PHITHAKROTCHANAKOON ◽  
Verawat CHAMPREDA ◽  
Sei-ichi AIBA ◽  
Kusol POOTANAKIT ◽  
Sutipa TANAPONGPIPAT
Keyword(s):  
Chain Length ◽  
Short Chain ◽  
Short Chain Length ◽  
Medium Chain ◽  
Medium Chain Length

scholarly journals Fructose-Based Production of Short-Chain-Length and Medium-Chain-Length Polyhydroxyalkanoate Copolymer by Arctic Pseudomonas sp. B14-6

Polymers ◽  
2021 ◽  
Vol 13 (9) ◽  
pp. 1398
Author(s):  
Tae-Rim Choi ◽  
Ye-Lim Park ◽  
Hun-Suk Song ◽  
Sun Mi Lee ◽  
Sol Lee Park ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Chain Length ◽  
Average Molecular Weight ◽  
Short Chain ◽  
Pha Synthase ◽  
Pseudomonas Sp ◽  
Short Chain Length ◽  
Medium Chain ◽  
Medium Chain Length ◽  
Pha Production

Arctic bacteria employ various mechanisms to survive harsh conditions, one of which is to accumulate carbon and energy inside the cell in the form of polyhydroxyalkanoate (PHA). Whole-genome sequencing of a new Arctic soil bacterium Pseudomonas sp. B14-6 revealed two PHA-production-related gene clusters containing four PHA synthase genes (phaC). Pseudomonas sp. B14-6 produced poly(6% 3-hydroxybutyrate-co-94% 3-hydroxyalkanoate) from various carbon sources, containing short-chain-length PHA (scl-PHA) and medium-chain-length PHA (mcl-PHA) composed of various monomers analyzed by GC-MS, such as 3-hydroxybutyrate, 3-hydroxyhexanoate, 3-hydroxyoctanoate, 3-hydroxydecanoate, 3-hydroxydodecenoic acid, 3-hydroxydodecanoic acid, and 3-hydroxytetradecanoic acid. By optimizing the PHA production media, we achieved 34.6% PHA content using 5% fructose, and 23.7% PHA content using 5% fructose syrup. Differential scanning calorimetry of the scl-co-mcl PHA determined a glass transition temperature (Tg) of 15.3 °C, melting temperature of 112.8 °C, crystallization temperature of 86.8 °C, and 3.82% crystallinity. In addition, gel permeation chromatography revealed a number average molecular weight of 3.6 × 104, weight average molecular weight of 9.1 × 104, and polydispersity index value of 2.5. Overall, the novel Pseudomonas sp. B14-6 produced a polymer with high medium-chain-length content, low Tg, and low crystallinity, indicating its potential use in medical applications.

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