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Purification, characterization, and properties of an alkaline protease produced by Serratia marcescens S3‐R1 inhabiting Korean ginseng rhizosphere
Author(s) -
Nam Myoung Soo,
Whang Kyung Sook,
Choi Seong Hyun,
Bae Hyoung Churl,
Kim Yoo Kyeong,
Park Young W
Publication year - 2013
Publication title -
journal of the science of food and agriculture
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 142
eISSN - 1097-0010
pISSN - 0022-5142
DOI - 10.1002/jsfa.6363
Subject(s) - chromatography , serratia marcescens , chemistry , protease , ammonium sulfate precipitation , ammonium sulfate , biochemistry , enzyme , escherichia coli , size exclusion chromatography , gene
BACKGROUND An alkaline protease produced by the Serratia marcescens S3‐R1 which inhabits in the Korean ginseng rhizosphere was investigated. The purposes of this study were to characterize and purify the bacterial enzyme by four different purification steps: precipitation of enzyme fraction by ammonium sulfate, loading the enzyme pellets on a DEAE –Sepharose anion‐exchange chromatograph, separation of the fraction containing enzyme activity by fast protein liquid Mono Q chromatography and identification of the single‐band fraction by sodium dodecyl sulfate–polyacrylamide gel electrophoresis, and then quantification of the single‐band fraction by reverse‐phase high‐performance liquid chromatography . RESULTS The molecular weight of the purified protease was estimated as 50 308 Da by matrix‐assisted laser desorption ionization time‐of‐flight analysis. The N‐terminal amino acid sequence of the protease was identified as Ala‐Val‐Thr‐Ile‐Glu‐Asp‐Ala‐Val‐Asp‐Asp, and the enzyme belongs to the metalloprotease family. The optimal activities of the protease occurred at pH 7–9 and a temperature 40 °C. The ranges of pH and thermal stability of the enzyme were at 7–10 and 30–40 °C, respectively . CONCLUSION The alkaline protease was successfully purified and characterized from the bacterium Serratia marcescens S3‐R1 , which has potential for industrial application, including milk protein hydrolysates. © 2013 Society of Chemical Industry