Poly-3-hydroxyalkanoate synthases from Pseudomonas putida U: substrate specificity and ultrastructural studies

被引:16
作者
Arias, Sagrario [1 ]
Sandoval, Angel [1 ]
Arcos, Mario [1 ]
Canedo, Librada M.
Maestro, Beatriz [2 ]
Sanz, Jesus M. [2 ]
Naharro, German [3 ]
Luengo, Jose M. [1 ]
机构
[1] Univ Leon, Dept Biol Mol, Fac Vet & Biol, E-24071 Leon, Spain
[2] Univ Miguel Hernandez, Inst Biol Mol & Celular, Elche 03202, Alicante, Spain
[3] Univ Leon, Dept Patol Anim Sanidad Anim, Fac Vet, E-24071 Leon, Spain
来源
MICROBIAL BIOTECHNOLOGY | 2008年 / 1卷 / 02期
关键词
D O I
10.1111/j.1751-7915.2007.00016.x
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The substrate specificity of the two polymerases (PhaC1 and PhaC2) involved in the biosynthesis of medium-chain-length poly-hydroxyalkanoates (mcl PHAs) in Pseudomonas putida U has been studied in vivo. For these kind of experiments, two recombinant strains derived from a genetically engineered mutant in which the whole pha locus had been deleted (P. putida U Delta pha) were employed. These bacteria, which expresses only phaC1 (P. putida U Delta pha pMC-phaC1) or only phaC2 (P. putida U Delta pha pMC-phaC2), accumulated different PHAs in function of the precursor supplemented to the culture broth. Thus, the P. putida U Delta pha pMC-phaC1 strain was able to synthesize several aliphatic and aromatic PHAs when hexanoic, heptanoic, octanoic decanoic, 5-phenylvaleric, 6-phenylhexanoic, 7-phenylheptanoic, 8-phenyloctanoic or 9-phenylnonanoic acid were used as precursors; the highest accumulation of polymers was observed when the precursor used were decanoic acid (aliphatic PHAs) or 6-phenylhexanoic acid (aromatic PHAs). However, although it synthesizes similar aliphatic PHAs (the highest accumulation was observed when hexanoic acid was the precursor) the other recombinant strain (P. putida U Delta pha pMC-phaC2) only accumulated aromatic PHAs when the monomer to be polymerized was 3-hydroxy-5-phenylvaleryl-CoA. The possible influence of the putative three-dimensional structures on the different catalytic behaviour of PhaC1 and PhaC2 is discussed.
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收藏
页码:170 / 176
页数:7
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