Purification and characterization of the E1 component of theClostridium magnum acetoin dehydrogenase enzyme system

1993 ◽  
Vol 64 (1) ◽  
pp. 9-15 ◽  
Author(s):  
Heidrun Lorenzl ◽  
Fred Bernd Oppermann ◽  
Bernhard Schmidt ◽  
Alexander Steinb�chel
Keyword(s):  
Enzyme System ◽  
Purification And Characterization ◽  
Dehydrogenase Enzyme

Purification and Characterization of a Newγ-Glutamylmethylamide-dissimilating Enzyme System fromMethylophagasp. AA-30

1995 ◽  
Vol 59 (4) ◽  
pp. 648-655 ◽  
Author(s):  
Toshio Kimura ◽  
Isao Sugahara ◽  
Katsuyuki Hanai ◽  
Tadashi Asahi
Keyword(s):  
Enzyme System ◽  
Purification And Characterization

scholarly journals Biochemical and molecular characterization of the Clostridium magnum acetoin dehydrogenase enzyme system.

1994 ◽  
Vol 176 (12) ◽  
pp. 3614-3630 ◽  
Author(s):  
N Krüger ◽  
F B Oppermann ◽  
H Lorenzl ◽  
A Steinbüchel
Keyword(s):  
Molecular Characterization ◽  
Enzyme System ◽  
Dehydrogenase Enzyme

scholarly journals Biochemical and Molecular Characterization of theBacillus subtilis Acetoin Catabolic Pathway

1999 ◽  
Vol 181 (12) ◽  
pp. 3837-3841 ◽  
Author(s):  
Min Huang ◽  
Fred Bernd Oppermann-Sanio ◽  
Alexander Steinbüchel
Keyword(s):  
Bacillus Subtilis ◽  
Carbon Source ◽  
Sole Carbon Source ◽  
Enzyme System ◽  
Gene Clusters ◽  
Catabolic Pathway ◽  
E Coli ◽  
Homologous Genes ◽  
Dehydrogenase Enzyme

ABSTRACT A recent study indicated that Bacillus subtiliscatabolizes acetoin by enzymes encoded by the acu gene cluster (F. J. Grundy, D. A. Waters, T. Y. Takova, and T. M. Henkin, Mol. Microbiol. 10:259–271, 1993) that are completely different from those in the multicomponent acetoin dehydrogenase enzyme system (AoDH ES) encoded by aco gene clusters found before in all other bacteria capable of utilizing acetoin as the sole carbon source for growth. By hybridization with a DNA probe covering acoA and acoB of the AoDH ES from Clostridium magnum, genomic fragments from B. subtilis harboring acoA, acoB,acoC, acoL, and acoR homologous genes were identified, and some of them were functionally expressed inE. coli. Furthermore, acoA was inactivated inB. subtilis by disruptive mutagenesis; these mutants were impaired to express PPi-dependent AoDH E1 activity to remove acetoin from the medium and to grow with acetoin as the carbon source. Therefore, acetoin is catabolized in B. subtilis by the same mechanism as all other bacteria investigated so far, leaving the function of the previously described acu genes obscure.

scholarly journals Thermophilic PHB depolymerase of Stenotrophomonas sp., an isolate from the plastic contaminated site is best purified on Octyl-Sepharose CL-4B

2019 ◽  
Author(s):  
R Z Sayyed ◽  
S J Wani ◽  
S S Shaikh ◽  
Helal F. Al-Harthi ◽  
Asad Syed ◽  
...  
Keyword(s):  
Column Chromatography ◽  
Large Scale ◽  
Enzyme System ◽  
Contaminated Sites ◽  
Contaminated Site ◽  
Purification And Characterization ◽  
Purification Method ◽  
Phb Depolymerase ◽  
Solvent Purification

AbstractThere are numerous reports on PHB depolymerases produced by a wide variety of microorganisms isolated from various habitats, however, reports on PHB depolymerase isolated from plastic contaminated sites are scares. Thermophilic PHB polymerase produced by isolates obtained from plastic contaminated sites is expected to have better relevance for its application in plastic/ bioplastic degradation. Although PHB has attracted commercial significance, the inefficient production and recovery methods, inefficient purification of PHB depolymerase and lack of ample knowledge on PHB degradation by PHB depolymerase have hampered its large scale commercialization. Therefore, to ensure the biodegradability of biopolymers, it becomes imperative to study the purification of the biodegrading enzyme system. We report the production, purification, and characterization of extracellular PHB depolymerase from Stenotrophomonas sp. RZS 7 isolated from a plastic contaminated site. The isolate produced extracellular poly-β-hydroxybutyrate (PHB) depolymerase in the mineral salt medium at 30oC during 4 days of incubation under shake flask condition. Purification of the enzyme was carried out by three different methods using PHB as a substrate. Purification of PHB depolymerase by ammonium salt precipitation, column chromatography, and solvent purification method was successfully carried out. Among the purification method tested, the enzyme was best purified by column chromatography on Octyl-Sepharose CL-4B column with maximum (0.7993 U/mg/ml) purification yield. The molecular weight of purified PHB depolymerase (40 kDa) closely resembled with PHB depolymerase of Aureobacterium saperdae.

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