Item request has been placed! ×
Item request cannot be made. ×
loading  Processing Request

Isolation and characterization of a heterologously expressed bacterial laccase from the anaerobe Geobacter metallireducens.

Item request has been placed! ×
Item request cannot be made. ×
loading   Processing Request
اقرأ على الانترنت اقرأ أكثر حفظ في قائمتي
  • معلومة اضافية
    • المصدر:
      Publisher: Springer International Country of Publication: Germany NLM ID: 8406612 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1432-0614 (Electronic) Linking ISSN: 01757598 NLM ISO Abbreviation: Appl Microbiol Biotechnol Subsets: MEDLINE
    • بيانات النشر:
      Original Publication: Berlin ; New York : Springer International, c1984-
    • الموضوع:
      Gene Expression*; Bacterial Proteins/*chemistry ; Bacterial Proteins/*isolation & purification ; Geobacter/*enzymology ; Laccase/*chemistry ; Laccase/*isolation & purification; Bacterial Proteins/genetics ; Bacterial Proteins/metabolism ; Cloning, Molecular ; Enzyme Stability ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Geobacter/chemistry ; Geobacter/genetics ; Hot Temperature ; Hydrogen-Ion Concentration ; Laccase/genetics ; Laccase/metabolism ; Substrate Specificity
    • نبذة مختصرة :
      Bioinformatics has revealed the presence of putative laccase genes in diverse bacteria, including extremophiles, autotrophs, and, interestingly, anaerobes. Integrity of laccase genes in anaerobes has been questioned, since laccases oxidize a variety of compounds using molecular oxygen as the electron acceptor. The genome of the anaerobe Geobacter metallireducens GS-15 contains five genes for laccase-like multicopper oxidases. In order to show whether one of the predicted genes encodes a functional laccase, the protein encoded by GMET_RS10855 was heterologously expressed in Escherichia coli cells. The His 6 -tagged enzyme (named GeoLacc) was purified to a large extent in the apoprotein, inactive form: incubation with CuSO 4 allowed a 43-fold increase of the specific activity yielding a metallo-enzyme. The purified enzyme oxidized some of the typical laccase substrates, including 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), syringaldazine, and 2,6-dimethoxyphenol (2,6-DMP), along with pyrogallol and K 4 [Fe(CN) 6 ]. Temperature optimum was 75 °C and pH optimum for ABTS and 2,6-DMP oxidation was ~ 6.0. As observed for other laccases, the enzyme was inhibited by halide anions and was sensitive to increasing concentrations of dimethyl sulfoxide and Tween-80. Notably, GeoLacc possesses a very high affinity for dioxygen: a similar activity was measured performing the reaction at air-saturated or microaerophilic conditions.
    • Grant Information:
      P4-0116 Javna Agencija za Raziskovalno Dejavnost RS; J4-4250 Javna Agencija za Raziskovalno Dejavnost RS
    • Contributed Indexing:
      Keywords: Anaerobic microorganisms; Bacterial laccase; Bioinformatics; Characterization; Heterologous expression
    • الرقم المعرف:
      0 (Bacterial Proteins)
      EC 1.10.3.2 (Laccase)
    • الموضوع:
      Date Created: 20180131 Date Completed: 20180814 Latest Revision: 20180814
    • الموضوع:
      20240829
    • الرقم المعرف:
      10.1007/s00253-018-8785-z
    • الرقم المعرف:
      29380032