Purification and characterization of branching specificity of a novel extracellular amylolytic enzyme from marine hyperthermophilic Rhodothermus marinus

Seong A. Yoon, Soo I. Ryu, Soo B. Lee, Tae W. Moon

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

An extracellular enzyme (RMEBE) possessing α-(1→4)-(1→6) -transferring activity was purified to homogeneity from Rhodothermus marinus by combination of ammonium sulfate precipitation, Q-Sepharose ion-exchange, and Superdex-200 gel filtration chromatographies, and preparative native polyacrylamide gel electrophoresis. The purified enzyme had an optimum pH of 6.0 and was highly thermostable with a maximal activity at 80°C. Its half-life was determined to be 73.7 and 16.7 min at 80 and 85°C, respectively. The enzyme was also halophilic and highly halotolerant up to about 2 M NaCl, with a maximal activity at 0.5 M. The substrate specificity of RMEBE suggested that it possesses partial characteristics of both glucan branching enzyme and neopullulanase. RMEBE clearly produced branched glucans from amylose, with partial α-(1→4)-hydrolysis of amylose and starch. At the same time, it hydrolyzed pullulan partly to panose, and exhibited α-(1→4)-(1→6)-transferase activity for small maltooligosaccharides, producing disproportionated α-(1→6)-branched maltooligosaccharides. The enzyme preferred maltopentaose and maltohexaose to smaller maltooligosaccharides for production of longer branched products. Thus, the results suggest that RMEBE might be applied for production of branched oligosaccharides from small maltodextrins at high temperature or even at high salinity.

Original languageEnglish
Pages (from-to)457-464
Number of pages8
JournalJournal of Microbiology and Biotechnology
Volume18
Issue number3
Publication statusPublished - 2008 Mar 28

Fingerprint

Rhodothermus
Purification
Enzymes
Amylose
Glucans
neopullulanase
1,4-alpha-Glucan Branching Enzyme
Native Polyacrylamide Gel Electrophoresis
Gels
Ion Exchange
Salinity
Ammonium Sulfate
Transferases
Substrate Specificity
Chromatography
Electrophoresis
Oligosaccharides
Starch
Sepharose
Gel Chromatography

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Microbiology

Cite this

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abstract = "An extracellular enzyme (RMEBE) possessing α-(1→4)-(1→6) -transferring activity was purified to homogeneity from Rhodothermus marinus by combination of ammonium sulfate precipitation, Q-Sepharose ion-exchange, and Superdex-200 gel filtration chromatographies, and preparative native polyacrylamide gel electrophoresis. The purified enzyme had an optimum pH of 6.0 and was highly thermostable with a maximal activity at 80°C. Its half-life was determined to be 73.7 and 16.7 min at 80 and 85°C, respectively. The enzyme was also halophilic and highly halotolerant up to about 2 M NaCl, with a maximal activity at 0.5 M. The substrate specificity of RMEBE suggested that it possesses partial characteristics of both glucan branching enzyme and neopullulanase. RMEBE clearly produced branched glucans from amylose, with partial α-(1→4)-hydrolysis of amylose and starch. At the same time, it hydrolyzed pullulan partly to panose, and exhibited α-(1→4)-(1→6)-transferase activity for small maltooligosaccharides, producing disproportionated α-(1→6)-branched maltooligosaccharides. The enzyme preferred maltopentaose and maltohexaose to smaller maltooligosaccharides for production of longer branched products. Thus, the results suggest that RMEBE might be applied for production of branched oligosaccharides from small maltodextrins at high temperature or even at high salinity.",
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Purification and characterization of branching specificity of a novel extracellular amylolytic enzyme from marine hyperthermophilic Rhodothermus marinus. / Yoon, Seong A.; Ryu, Soo I.; Lee, Soo B.; Moon, Tae W.

In: Journal of Microbiology and Biotechnology, Vol. 18, No. 3, 28.03.2008, p. 457-464.

Research output: Contribution to journalArticle

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