Expression and Characterization of (R)-specific Enoyl Coenzyme A Hydratase Involved in Polyhydroxyalkanoate Biosynthesis by Aeromonas Caviae

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 1998 Feb 11

PMID 9457873

Citations 40

Authors

T Fukui

N Shiomi

Y Doi

Affiliations

Soon will be listed here.

Abstract

Complementation analysis of a polyhydroxyalkanoate (PHA)-negative mutant of Aeromonas caviae proved that ORF3 in the pha locus (a 402-bp gene located downstream of the PHA synthase gene) participates in PHA biosynthesis on alkanoic acids, and the ORF3 gene is here referred to as phaJ(Ac). Escherichia coli BL21(DE3) carrying phaJ(Ac). under the control of the T7 promoter overexpressed enoyl coenzyme A (enoyl-CoA) hydratase, which was purified by one-step anion-exchange chromatography. The N-terminal amino acid sequence of the purified hydratase corresponded to the amino acid sequence deduced from the nucleotide sequence of phaJ(Ac) except for the initial Met residue. The enoyl-CoA hydratase encoded by phaJ(Ac) exhibited (R)-specific hydration activity toward trans-2-enoyl-CoA with four to six carbon atoms. These results have demonstrated that (R)-specific hydration of 2-enoyl-CoA catalyzed by the translated product of phaJ(Ac) is a channeling pathway for supplying (R)-3-hydroxyacyl-CoA monomer units from fatty acid beta-oxidation to poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) biosynthesis in A. caviae.

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References

Timm A, Steinbuchel A . Formation of polyesters consisting of medium-chain-length 3-hydroxyalkanoic acids from gluconate by Pseudomonas aeruginosa and other fluorescent pseudomonads. Appl Environ Microbiol. 1990; 56(11):3360-7. PMC: 184954. DOI: 10.1128/aem.56.11.3360-3367.1990. View

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

Schlegel H, LAFFERTY R, Krauss I . The isolation of mutants not accumulating poly-beta-hydroxybutyric acid. Arch Mikrobiol. 1970; 71(3):283-94. DOI: 10.1007/BF00410161. View

Huijberts G, Eggink G, De Waard P, Huisman G, Witholt B . Pseudomonas putida KT2442 cultivated on glucose accumulates poly(3-hydroxyalkanoates) consisting of saturated and unsaturated monomers. Appl Environ Microbiol. 1992; 58(2):536-44. PMC: 195281. DOI: 10.1128/aem.58.2.536-544.1992. View

Huisman G, de Leeuw O, Eggink G, Witholt B . Synthesis of poly-3-hydroxyalkanoates is a common feature of fluorescent pseudomonads. Appl Environ Microbiol. 1989; 55(8):1949-54. PMC: 202985. DOI: 10.1128/aem.55.8.1949-1954.1989. View

Davison J, Heusterspreute M, Chevalier N, Brunel F . Vectors with restriction site banks. V. pJRD215, a wide-host-range cosmid vector with multiple cloning sites. Gene. 1987; 51(2-3):275-80. DOI: 10.1016/0378-1119(87)90316-7. View

Peoples O, Sinskey A . Poly-beta-hydroxybutyrate (PHB) biosynthesis in Alcaligenes eutrophus H16. Identification and characterization of the PHB polymerase gene (phbC). J Biol Chem. 1989; 264(26):15298-303. View

Slater S, Voige W, Dennis D . Cloning and expression in Escherichia coli of the Alcaligenes eutrophus H16 poly-beta-hydroxybutyrate biosynthetic pathway. J Bacteriol. 1988; 170(10):4431-6. PMC: 211473. DOI: 10.1128/jb.170.10.4431-4436.1988. View

Leaf T, Peterson M, Stoup S, Somers D, Srienc F . Saccharomyces cerevisiae expressing bacterial polyhydroxybutyrate synthase produces poly-3-hydroxybutyrate. Microbiology (Reading). 1996; 142 ( Pt 5):1169-1180. DOI: 10.1099/13500872-142-5-1169. View

10.

Steinbuchel A, Hustede E, Liebergesell M, Pieper U, Timm A, Valentin H . Molecular basis for biosynthesis and accumulation of polyhydroxyalkanoic acids in bacteria. FEMS Microbiol Rev. 1992; 9(2-4):217-30. DOI: 10.1111/j.1574-6968.1992.tb05841.x. View

11.

Williams M, Rahn J, Sherman D . Production of a polyhydroxyalkanoate biopolymer in insect cells with a modified eucaryotic fatty acid synthase. Appl Environ Microbiol. 1996; 62(7):2540-6. PMC: 168036. DOI: 10.1128/aem.62.7.2540-2546.1996. View

12.

Anderson A, DAWES E . Occurrence, metabolism, metabolic role, and industrial uses of bacterial polyhydroxyalkanoates. Microbiol Rev. 1990; 54(4):450-72. PMC: 372789. DOI: 10.1128/mr.54.4.450-472.1990. View

13.

Brandl H, Gross R, Lenz R, Fuller R . Pseudomonas oleovorans as a Source of Poly(beta-Hydroxyalkanoates) for Potential Applications as Biodegradable Polyesters. Appl Environ Microbiol. 1988; 54(8):1977-82. PMC: 202789. DOI: 10.1128/aem.54.8.1977-1982.1988. View

14.

Friedrich B, Hogrefe C, Schlegel H . Naturally occurring genetic transfer of hydrogen-oxidizing ability between strains of Alcaligenes eutrophus. J Bacteriol. 1981; 147(1):198-205. PMC: 216026. DOI: 10.1128/jb.147.1.198-205.1981. View

15.

De Waard P, van der Wal H, Huijberts G, Eggink G . Heteronuclear NMR analysis of unsaturated fatty acids in poly(3-hydroxyalkanoates). Study of beta-oxidation in Pseudomonas putida. J Biol Chem. 1993; 268(1):315-9. View

16.

Poirier Y, Nawrath C, Somerville C . Production of polyhydroxyalkanoates, a family of biodegradable plastics and elastomers, in bacteria and plants. Biotechnology (N Y). 1995; 13(2):142-50. DOI: 10.1038/nbt0295-142. View

17.

Deng W, Nickoloff J . Site-directed mutagenesis of virtually any plasmid by eliminating a unique site. Anal Biochem. 1992; 200(1):81-8. DOI: 10.1016/0003-2697(92)90280-k. View

18.

Brandl H, Knee Jr E, Fuller R, Gross R, Lenz R . Ability of the phototrophic bacterium Rhodospirillum rubrum to produce various poly (beta-hydroxyalkanoates): potential sources for biodegradable polyesters. Int J Biol Macromol. 1989; 11(1):49-55. DOI: 10.1016/0141-8130(89)90040-8. View

19.

Fukui T, Doi Y . Cloning and analysis of the poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) biosynthesis genes of Aeromonas caviae. J Bacteriol. 1997; 179(15):4821-30. PMC: 179330. DOI: 10.1128/jb.179.15.4821-4830.1997. View

20.

Stern J, DEL CAMPILLO A, Raw I . Enzymes of fatty acid metabolism. I. General introduction; crystalline crotonase. J Biol Chem. 1956; 218(2):971-83. View