[Expression, thermal stability modification and application in PHB degradation of polyhydroxyalkanoate depolymerase from Thermomonospora umbrina].
Sheng Wu Gong Cheng Xue Bao
; 39(8): 3351-3363, 2023 Aug 25.
Article
em Zh
| MEDLINE
| ID: mdl-37622365
ABSTRACT
Polyhydroxyalkanoate depolymerase (PHAD) can be used for the degradation and recovery of polyhydroxyalkanoate (PHA). In order to develop a PHAD with good stability under high temperature, PHAD from Thermomonospora umbrina (TumPHAD) was heterelogously expressed in Escherichia coli BL21(DE3). At the same time, a mutant A190C/V240C with enhanced stability was obtained via rational design of disulfide bonds. Characterization of enzymatic properties showed that the mutant A190C/V240C had an optimum temperature of 60 â, which was 20 â higher than that of the wild type. The half-life at 50 â was 7 hours, at 50 â which was 21 times longer than that of the wild type. The mutant A190C/V240C was used for the degradation of polyhydroxybutyrate (PHB), one of the typical PHA. At 50 â, the degradation rate of PHB being treated for 2 hours and 12 hours was 2.1 times and 3.8 times higher than that of the wild type, respectively. The TumPHAD mutant A190C/V240C obtained in this study shows tolerance to high temperature resistance, good thermal stability and strong PHB degradation ability, which may facilitate the degradation and recovery of PHB.
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MEDLINE
Assunto principal:
Actinomycetales
/
Poli-Hidroxialcanoatos
Idioma:
Zh
Ano de publicação:
2023
Tipo de documento:
Article