In this work, sucrose utilizing microbes from soil were screened to evaluate their ability for accumulation of biopolymer of polyhydroxyalkanoate (PHA). Among 72 isolates were transferred to mineral salt medium (MS...In this work, sucrose utilizing microbes from soil were screened to evaluate their ability for accumulation of biopolymer of polyhydroxyalkanoate (PHA). Among 72 isolates were transferred to mineral salt medium (MSM), 33 strains can be grown on sucrose agar medium. However, only one strain showed a strong black color for Sudan Black and gave positive result for Nile blue A. Identification by 16S rDNA nucleotide sequence homology of the isolate showed very closely to Hydrogenophaga sp. (99% identify). To consider PHA production, the isolate was grown in the medium containing sucrose as a sole carbon under controlled conditions of 35 ℃ and at pH 7. Maximum dry cell weight (DCW) and PHA production were obtained at 3.61 g/L and 2.41 g/L after 36 and 42 h batch fermentation. Sucrose uptake measured in term of total organic carbon (TOC) showed at 14.73 g within 48 h. The highest PHA was 68.15% (gPHA/gDCW) giving maximum PHA yield (YP/s) of 0.17 (gPHA/gs ) and a productivity of 0.057 gPHA/L.h. This highlights the potential of microbial resources in soil environment and may be an exploitable application for the industrial production of PHA.展开更多
Synthetic biology promises to simplify the construction of metabolic pathways by assembling the de- tached modules of the whole pathway. This gives new approaches for the microbial production of industrial products su...Synthetic biology promises to simplify the construction of metabolic pathways by assembling the de- tached modules of the whole pathway. This gives new approaches for the microbial production of industrial products such as polyhydroxyalkanoates (PHA). In this study, to produce poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) by Pseudomonas stutzeri 1317 from unrelated carbon sources such as glucose, the phaCl-phaZ-phaC2 operon of P. stutzeri 1317 was knocked out to generate the PHA deficient mutant P. stutzeri 1317LF. Then three modules containing phaCahAReBRe, phaCahBReGep and phaCAhPah were introduced into P. stutzeri 1317LF separately The shake flask results indicated that the precursor supply and PHA synthase activity were the vital factors for the PHBHHx accumulation of P. stutzeri 1317LF. Furthermore, the PHBHHx accumulation of the recombinants from different carbon resources were performed. The highest PHBHHx content was 23.7% (by mass) with 58.6% (by mole) 3HB fraction. These results provide basis for further improving the PHBHHx accumulation of P. stutzeri from unrelated carbon sources.展开更多
Microbial synthesis of functional polymers has become increasingly important for industrial biotechnology. For the first time, it became possible to synthesize controllable composition of poly(3-hydroxyalkanoate) (...Microbial synthesis of functional polymers has become increasingly important for industrial biotechnology. For the first time, it became possible to synthesize controllable composition of poly(3-hydroxyalkanoate) (P3HA) consisting of 3-hydroxydodec- anoate (3HDD) and phenyl group on the side-chain when chromosome of Pseudomonas entomophila was edited to weaken its t-oxidation. Cultured in the presence of 5-phenylvaleric acid (PVA), the edited P. entomophila produced only homopolymer poly(3-hydroxy-5-phenylvalerate) or P(3HPhV). While copolyesters P(3HPhV-co-3HDD) of 3-hydroxy-5-phenylvalerate (3HPhV) and 3-hydroxydodecanoate (3HDD) were synthesized when the strain was grown on mixtures of PVA and dodecanoic acid (DDA). Compositions of 3HPhV in P(3HPhV-co-3HDD) were controllable ranging from 3% to 32% depending on DDDA/PVA ratios. Nuclear magnetic resonance (NMR) spectra clearly indicated that the polymers were homopolymer of P(3HPhV) and random copolymers of 3HPhV and 3HDD. Their mechanical and thermal properties varied dramatically de- pending on the monomer ratios. Our results demonstrated the possibility to produce tailor-made, novel functional PHA using the chromosome edited P. entomophila.展开更多
Halophilic archaea (haloarchaea) inhabit hypersaline environments,tolerating extreme salinity,low oxygen and nutrient availability,and in some cases,high pH (soda lakes) and irradiation (saltern ponds).Membrane-associ...Halophilic archaea (haloarchaea) inhabit hypersaline environments,tolerating extreme salinity,low oxygen and nutrient availability,and in some cases,high pH (soda lakes) and irradiation (saltern ponds).Membrane-associated proteins of haloarchaea,such as surface layer (S-layer) proteins,transporters,retinal proteins,and internal organellar membrane proteins including intracellular gas vesicle proteins and those associated with polyhydroxyalkanoate (PHA) granules,contribute greatly to their environmental adaptations.This review focuses on these haloarchaeal cellular and organellar membrane-associated proteins,and provides insight into their physiological significance and biotechnological potential.展开更多
文摘In this work, sucrose utilizing microbes from soil were screened to evaluate their ability for accumulation of biopolymer of polyhydroxyalkanoate (PHA). Among 72 isolates were transferred to mineral salt medium (MSM), 33 strains can be grown on sucrose agar medium. However, only one strain showed a strong black color for Sudan Black and gave positive result for Nile blue A. Identification by 16S rDNA nucleotide sequence homology of the isolate showed very closely to Hydrogenophaga sp. (99% identify). To consider PHA production, the isolate was grown in the medium containing sucrose as a sole carbon under controlled conditions of 35 ℃ and at pH 7. Maximum dry cell weight (DCW) and PHA production were obtained at 3.61 g/L and 2.41 g/L after 36 and 42 h batch fermentation. Sucrose uptake measured in term of total organic carbon (TOC) showed at 14.73 g within 48 h. The highest PHA was 68.15% (gPHA/gDCW) giving maximum PHA yield (YP/s) of 0.17 (gPHA/gs ) and a productivity of 0.057 gPHA/L.h. This highlights the potential of microbial resources in soil environment and may be an exploitable application for the industrial production of PHA.
基金Supported by the National lqatural Science Foundation of China (31260015), Natural Science Foundation of Qinghai Province (2012-Z-919Q), the Extramural Project from State Key Laboratory for Agrobiotechnology (2012SKLAB06-5) and the Research Funds for Young Project of Qinghal University (2011-QYY-1).
文摘Synthetic biology promises to simplify the construction of metabolic pathways by assembling the de- tached modules of the whole pathway. This gives new approaches for the microbial production of industrial products such as polyhydroxyalkanoates (PHA). In this study, to produce poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) by Pseudomonas stutzeri 1317 from unrelated carbon sources such as glucose, the phaCl-phaZ-phaC2 operon of P. stutzeri 1317 was knocked out to generate the PHA deficient mutant P. stutzeri 1317LF. Then three modules containing phaCahAReBRe, phaCahBReGep and phaCAhPah were introduced into P. stutzeri 1317LF separately The shake flask results indicated that the precursor supply and PHA synthase activity were the vital factors for the PHBHHx accumulation of P. stutzeri 1317LF. Furthermore, the PHBHHx accumulation of the recombinants from different carbon resources were performed. The highest PHBHHx content was 23.7% (by mass) with 58.6% (by mole) 3HB fraction. These results provide basis for further improving the PHBHHx accumulation of P. stutzeri from unrelated carbon sources.
基金supported by the National High Technology Research and Development Program of China(2012AA023102 to Liu Lei,Guo Kai and Wu Qiong)the National Basic Research Program of China(2012CB725201 to Chen GuoQiang and Chen JinChun,2012CB725204 to Guo Kai and Wu Qiong)National Natural Science Foundation of China(31270146 to Chen GuoQiang)
文摘Microbial synthesis of functional polymers has become increasingly important for industrial biotechnology. For the first time, it became possible to synthesize controllable composition of poly(3-hydroxyalkanoate) (P3HA) consisting of 3-hydroxydodec- anoate (3HDD) and phenyl group on the side-chain when chromosome of Pseudomonas entomophila was edited to weaken its t-oxidation. Cultured in the presence of 5-phenylvaleric acid (PVA), the edited P. entomophila produced only homopolymer poly(3-hydroxy-5-phenylvalerate) or P(3HPhV). While copolyesters P(3HPhV-co-3HDD) of 3-hydroxy-5-phenylvalerate (3HPhV) and 3-hydroxydodecanoate (3HDD) were synthesized when the strain was grown on mixtures of PVA and dodecanoic acid (DDA). Compositions of 3HPhV in P(3HPhV-co-3HDD) were controllable ranging from 3% to 32% depending on DDDA/PVA ratios. Nuclear magnetic resonance (NMR) spectra clearly indicated that the polymers were homopolymer of P(3HPhV) and random copolymers of 3HPhV and 3HDD. Their mechanical and thermal properties varied dramatically de- pending on the monomer ratios. Our results demonstrated the possibility to produce tailor-made, novel functional PHA using the chromosome edited P. entomophila.
基金supported by the National High Technology Research and Development Program of China (Grant No. 2010AA09Z401)National Natural Science Foundation of China (Grant Nos.30621005,30830004, and 30925001)+1 种基金Chinese Academy of Sciences (KSCXZEW-G-2-4)supported by the US National Aeronautics and Space Administration (Grant No.NNX10AP47G awarded to S. DasSarma)
文摘Halophilic archaea (haloarchaea) inhabit hypersaline environments,tolerating extreme salinity,low oxygen and nutrient availability,and in some cases,high pH (soda lakes) and irradiation (saltern ponds).Membrane-associated proteins of haloarchaea,such as surface layer (S-layer) proteins,transporters,retinal proteins,and internal organellar membrane proteins including intracellular gas vesicle proteins and those associated with polyhydroxyalkanoate (PHA) granules,contribute greatly to their environmental adaptations.This review focuses on these haloarchaeal cellular and organellar membrane-associated proteins,and provides insight into their physiological significance and biotechnological potential.