Gene Cloning and Characterization of a Poly(<Emphasis Type="SmallCaps">l</Emphasis>-Lactic Acid) Depolymerase from <Emphasis Type="Italic">Pseudomonas</Emphasis> sp. Strain DS04-T |
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| Authors: | Zhan-Yong Wang Fan Li Zi-Qi Guo Yan Wang Shan Chen |
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| Institution: | (1) School of Life Sciences, Northeast Normal University, Changchun, 130024, People’s Republic of China;(2) School of Environmental and Biological Engineering, Liaoning Shihua University, Fushun, 113001, People’s Republic of China |
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| Abstract: | A gene encoding a poly(l-lactic acid) (PLA) depolymerase from Pseudomonas sp. strain DS04-T was cloned and overexpressed in Escherichia coli. The recombinant PLA depolymerase with a molecular weight of 19.2 kDa was purified to homogeneity. The optimum pH and temperature
of the PLA depolymerase are 8.5 and 60 °C, respectively. K+, Ca2+ and Ni2+ enhance the enzyme activity, while Na+, Zn2+, Mg2+, Cu2+, Fe2+, Mn2+ and Co2+ inhibit it. The inhibition of different chemicals on the PLA depolymerase activity were examined, in which EDTA was found
to have a significantly inhibitory effect. The main degradation product of the depolymerase is identified as lactic acid monomer
by mass spectrometric analysis. Physicochemical properties, substrate specificity and sequence analysis indicated that PME
is a new type of PLA depolymerase. |
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