Localization and inactivation of DNase activity in Clostridium pasteurianum NRRL-B598

Auranee Boonsermsuwong, Hans P. Blaschek

Research output: Contribution to journalArticlepeer-review

Abstract

Each of four cell fractions of Clostridium pasteurianum NRRL-B598 contained DNase. The culture supernatant contained 31.3% of the total cellular DNase activity, while the cell wash, cell-wall compartmentalized and intracellular fractions contained 27.8, 27.8 and 13.1%, respectively. This enzyme was thermoresistant and heat treatment at various stages of the plasmid isolation protocol reduced chromosomal smearing, but did not improve plasmid recovery. The use of a new density gradient material (cesium trifluoroacetate; CsTFA) known to strongly inactivate DNase was compared to standard isopycnic centrifugation with cesium chloride. The consistent recovery of a 2.3 MDa plasmid only in the presence of CsTFA suggests that DNase continues to be a problem during ultracentrifugation as well as throughout the cleared lysate protocol.

Original languageEnglish (US)
Pages (from-to)265-270
Number of pages6
JournalJournal of Industrial Microbiology
Volume1
Issue number4
DOIs
StatePublished - Dec 1 1986

Keywords

  • Clostridium pasteurianum
  • CsTFA
  • DNase
  • Localization
  • Plasmid recovery

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology

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