Purification and characterization of a maltopentaose-producing amylase from Bacillus megaterium KSM B-404

Je Won Park, Byung Ju Kim, Jae Woo Lee, Young Bae Kim

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

An amylase that hydrolyzes starch into maltopentaose as a main product was found in the culture supernatant of a strain of Bacillus megaterium KSM B-404 isolated from local soil. The enzyme was purified 129-fold by ammonium sulfate precipitation, DEAE-Toyopearl and Superdex 75 HR 10/30 column using a FPLC system. The molecular weight of the amylase was determined as about 68 kDa by using SDS-PAGE. Optimum pH and temperature of amylase were found to be 50°C and pH 6.0∼7.0, respectively. The enzyme was stable up to 60°C by addition of Ca2+ and its pH stability was in the range of 6.0∼10.0. The activity of enzyme was inhibited by Cu2+, Hg 2+, and Fe3+ and maintained by Ca2+ and Mg 2+. EDTA and pCMB also showed inhibitory effect to the enzyme. TLC and HPLC analysis of the products of the enzyme reaction showed the presence of maltopentaose (52%), maltotriose (25%), maltose (11%), glucose, and maltotetraose in the starch hydrolysates.

Original languageEnglish
Pages (from-to)352-358
Number of pages7
JournalKorean Journal of Microbiology and Biotechnology
Volume30
Issue number4
Publication statusPublished - 2002 Dec 1

Fingerprint

Bacillus megaterium
Amylases
Enzymes
Starch
Maltose
Ammonium Sulfate
Edetic Acid
Polyacrylamide Gel Electrophoresis
Soil
Molecular Weight
High Pressure Liquid Chromatography
maltopentaose
Glucose
Temperature

Keywords

  • Amylase
  • Bacillus megaterium KSM B-404
  • Maltopentaose

ASJC Scopus subject areas

  • Biotechnology
  • Microbiology
  • Applied Microbiology and Biotechnology

Cite this

Purification and characterization of a maltopentaose-producing amylase from Bacillus megaterium KSM B-404. / Park, Je Won; Kim, Byung Ju; Lee, Jae Woo; Kim, Young Bae.

In: Korean Journal of Microbiology and Biotechnology, Vol. 30, No. 4, 01.12.2002, p. 352-358.

Research output: Contribution to journalArticle

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