Purification and properties of an intracellular 3-hydroxybutyrate-oligomer hydrolase (PhaZ2) in Ralstonia eutropha H16 and its identification as a novel intracellular poly(3-hydroxybutyrate) depolymerase.

Journal of Bacteriology
Teruyuki KobayashiT Saito

Abstract

An intracellular 3-hydroxybutyrate (3HB)-oligomer hydrolase (PhaZ2(Reu)) of Ralstonia eutropha was purified from Escherichia coli harboring a plasmid containing phaZ2(Reu). The purified enzyme hydrolyzed linear and cyclic 3HB-oligomers. Although it did not degrade crystalline poly(3-hydroxybutyrate) (PHB), the purified enzyme degraded artificial amorphous PHB at a rate similar to that of the previously identified intracellular PHB (iPHB) depolymerase (PhaZ1(Reu)). The enzyme appeared to be an endo-type hydrolase, since it actively hydrolyzed cyclic 3HB-oligomers. However, it degraded various linear 3HB-oligomers and amorphous PHB in the fashion of an exo-type hydrolase, releasing one monomer unit at a time. PhaZ2 was found to bind to PHB inclusion bodies and as a soluble enzyme to cell-free supernatant fractions in R. eutropha; in contrast, PhaZ1 bound exclusively to the inclusion bodies. When R. eutropha H16 was cultivated in a nutrient-rich medium, the transient deposition of PHB was observed: the content of PHB was maximized in the log growth phase (12 h, ca. 14% PHB of dry cell weight) and decreased to a very low level in the stationary phase (ca. 1% of dry cell weight). In each phaZ1-null mutant and phaZ2-null mutant, the ...Continue Reading

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Citations

Feb 9, 2012·Applied Microbiology and Biotechnology·Ulrike BrandtAlexander Steinbüchel
Mar 15, 2008·Antonie van Leeuwenhoek·Akiko SugimotoTerumi Saito
Sep 8, 2006·Applied and Environmental Microbiology·Birgit Gebauer, Dieter Jendrossek
Dec 25, 2007·Applied and Environmental Microbiology·Keiichi UchinoDieter Jendrossek
Mar 10, 2009·Journal of Bacteriology·Dieter Jendrossek
Jul 21, 2005·Journal of Bacteriology·Teruyuki KobayashiTerumi Saito
Jun 15, 2005·Annual Review of Biochemistry·Joanne StubbePinghua Liu
Nov 12, 2013·Applied and Environmental Microbiology·Daniel Pfeiffer, Dieter Jendrossek
Apr 23, 2015·Bioscience, Biotechnology, and Biochemistry·Pabitra BhagowatiSurajit Das
Mar 29, 2006·Journal of Biochemistry·Keiichi Uchino, Terumi Saito
Apr 7, 2020·Advanced Materials·So Young ChoiSang Yup Lee
May 26, 2018·World Journal of Microbiology & Biotechnology·Gurusamy Kutralam-Muniasamy, Fermín Peréz-Guevara
Nov 1, 2020·International Journal of Biological Macromolecules·Salma ShahidZill-I-Huma Nazli
Aug 5, 2021·Applied Biochemistry and Biotechnology·Shivam BholaPradeep Kumar
Oct 30, 2007·Journal of Bioscience and Bioengineering·Keiichi UchinoTerumi Saito

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