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

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 journal › Article › peer-review

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 language	English
Pages (from-to)	352-358
Number of pages	7
Journal	Korean Journal of Microbiology and Biotechnology
Volume	30
Issue number	4
Publication status	Published - 2002 Dec

Keywords

Amylase
Bacillus megaterium KSM B-404
Maltopentaose

ASJC Scopus subject areas

Biotechnology
Microbiology
Applied Microbiology and Biotechnology

Cite this

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title = "Purification and characterization of a maltopentaose-producing amylase from Bacillus megaterium KSM B-404",

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.",

keywords = "Amylase, Bacillus megaterium KSM B-404, Maltopentaose",

author = "Park, {Je Won} and Kim, {Byung Ju} and Lee, {Jae Woo} and Kim, {Young Bae}",

year = "2002",

month = dec,

language = "English",

volume = "30",

pages = "352--358",

journal = "Korean Journal of Microbiology and Biotechnology",

issn = "1598-642X",

publisher = "Korean Society for Microbiolog and Biotechnology",

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TY - JOUR

T1 - Purification and characterization of a maltopentaose-producing amylase from Bacillus megaterium KSM B-404

AU - Park, Je Won

AU - Kim, Byung Ju

AU - Lee, Jae Woo

AU - Kim, Young Bae

PY - 2002/12

Y1 - 2002/12

N2 - 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.

AB - 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.

KW - Amylase

KW - Bacillus megaterium KSM B-404

KW - Maltopentaose

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M3 - Article

AN - SCOPUS:0141995708

SN - 1598-642X

VL - 30

SP - 352

EP - 358

JO - Korean Journal of Microbiology and Biotechnology

JF - Korean Journal of Microbiology and Biotechnology

IS - 4

ER -

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

Abstract

Keywords

ASJC Scopus subject areas

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