Alpha-Amylase from Persimmon Honey: Purification and Characterization

The α-amylase was extracted from pure persimmon honey and purified by DEAE-Toyopearl 650M, CM-Toyopearl 650M, and Toyopearl HW-55F column chromatographies. Molecular weight of purified enzyme was estimated to be about 58 kDa by Toyopearl HW-55F gel chromatography and SDS-PAGE, respectively suggested that the purified enzyme was a monomer. Optimum pH of the enzyme was 6.0−7.0 and optimum temperature 40°C. The enzyme was extremely inactivated at pH was higher than 7.0 or lower than 5.0. Heat inactivation occurred at 40°C. This enzyme activated by Ca 2+ , Mn2+, PCMB, and DTNB, but inhibited by Ba2+, Fe3+, Hg2+, Mg2+, and iodoacetic acid. The purified enzyme was of α -type by TLC analysis. The relative rate of hydrolysis of the polymeric substance decreased with decreasing percentage of α -1,4-linkages and with increasing percentage of α -1,6-linkages in substrate similar to the results from commercially available honey.

[1]  S. Babacan,et al.  Characterization of honey amylase. , 2007, Journal of food science.

[2]  S. Babacan,et al.  Purification of Amylase from Honey , 2006 .

[3]  C. E. Lupano,et al.  Crystallization of Honey at −20°C , 2006 .

[4]  P. Sen,et al.  Purification and some properties of α-amylase from post-harvest Pachyrhizus erosus L. tuber , 2006 .

[5]  E. Emanuilova,et al.  Purification and properties of heat stable α-amylase from Bacillus brevis , 1989, Applied Microbiology and Biotechnology.

[6]  C. Vieille,et al.  Pyrococcus furiosus α-Amylase Is Stabilized by Calcium and Zinc† , 2002 .

[7]  T. Nakajima,et al.  Characterization of the functional module responsible for the low temperature optimum of a rice -amylase (Amy 3E) , 2002 .

[8]  W. Miller,et al.  Purification and characterization of an endoamylase from tulip (Tulipa gesneriana) bulbs , 2000 .

[9]  R. García-Villanova,et al.  Geographical discrimination of honeys by using mineral composition and common chemical quality parameters , 2000 .

[10]  T. Kubo,et al.  Expression of amylase and glucose oxidase in the hypopharyngeal gland with an age-dependent role change of the worker honeybee (Apis mellifera L.). , 1999, European journal of biochemistry.

[11]  C. Romero Investigación Agraria en Castilla-La Mancha. , 1999 .

[12]  C. Buesa,et al.  Purification and characterization of a new alpha-amylase of intermediate thermal stability from the yeast Lipomyces kononenkoae. , 1995, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[13]  S. Freer Purification and characterization of the extracellular alpha-amylase from Streptococcus bovis JB1 , 1993, Applied and environmental microbiology.

[14]  G. Goma,et al.  Purification and characterization of the extracellular alpha-amylase from Clostridium acetobutylicum ATCC 824 , 1991, Applied and environmental microbiology.

[15]  J. Baker Purification and partial characterization of α-Amylase allozymes from the lesser grain borer, Rhyzopertha dominica☆☆☆ , 1991 .

[16]  Wulf Crueger,et al.  Biotechnology: A textbook of industrial microbiology , 1984 .

[17]  J. E. Alter,et al.  Purification and Characterization of a Thermostable alpha‐Amylase from Bacillus licheniformis , 1979 .

[18]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[19]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.