Purification and characterization of poly(3-hydroxylbutyrate) depolymerase from a fungal isolate, Emericellopsis minima W2

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Title
Purification and characterization of poly(3-hydroxylbutyrate) depolymerase from a fungal isolate, Emericellopsis minima W2
Author(s)
Do Young Kim; Ji Hye Yun; Hyung Woo Kim; Kyung Sook Bae; Young Ha Rhee
Bibliographic Citation
Journal of Microbiology, vol. 40, no. 2, pp. 129-133
Publication Year
2002
Abstract
The fungus, Emericellopsis minima W2, capable of degrading poly(3-hydroxybutyrate) (PHB) was isolated from a waste water sample. Production of the PHB depolymerase from E. minima W2 (PhaZEmi) was significantly repressed in the presence of glucose. PhaZEmi was purified by column chromatography on Octyl-Sepharose CL-4B and Sephadex G-100. The molecular mass of the PhaZEmi, which consisted of a single polypeptide chain, was estimated to be 48.0 kDa by SDS-PAGE and its pI value was 4.4. The maximum activity of the PhaZEmi was observed at pH 9.0 and 55°C. It was significantly inactivated by 1 mM dithiothreitol, 2 mM diisopropyl fluorophosphate, 0.1 mM Tween 80, and 0.1 mM Triton X-100, but insensitive to phenylmethylsulfonyl fluoride and N-ethylmaleimide. The PhaZEmi efficiently hydrolyzed PHB and its copolyester with 30 mol% 3-hydroxyvalerate, but did not act on poly(3-hydroxyoctanoate). It also hydrolyzed p-nitrophenylacetate and p-nitrophenylbutyrate but hardly affected the longer-chain forms. The main hydrolysis product of PHB was identified as a dimer of 3-hydroxybutyrate.
Keyword
emericellopsis minimaPHB depolymerasepoly(3-hydroxyalkanoate)poly(3-hydroxybutyrate)
ISSN
1225-8873
Publisher
Microbiological Society Korea
Type
Article
Appears in Collections:
Division of Biomaterials Research > Industrial Bio-materials Research Center > 1. Journal Articles
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