title: Extracellular production of beta-amylase by a halophilic isolate, Halobacillus sp. LY9.
abstract: A moderately halophilic strain LY9 with high amylolytic activity was isolated from soil sample obtained from Yuncheng, China. Biochemical and physiological characterization along with 16S rRNA sequence analysis placed the isolate in the genus Halobacillus. Amylase production started from the post-exponential phase of bacterial growth and reached a maximum level during the early-stationary phase. The isolate LY9 was found to secrete the amylase, the production of which depended on the salinity of the growth medium. Maximum amylase production was observed in the presence of 10% KCl or 10% NaCl. Maltose was the main product of soluble starch hydrolysis, indicating a β-amylase activity. The enzyme showed optimal activity at 60°C, pH 8.0, and 10-12.5% of NaCl. It was highly active over broad temperature (50-70°C), NaCl concentration (5.0-20.0%), and pH (4.0-12.0) ranges, indicating its thermoactive and alkali-stable nature. However, activity dropped off dramatically at low NaCl concentrations, showing the amylase was halophilic. Ca(2+) was found to stimulate the β-amylase activity, whereas ethylenediaminetetraacetic acid (EDTA), phenylarsine oxide (PAO), and diethyl pyrocarbonate (DEPC) strongly inhibited the enzyme, indicating it probably was a metalloenzyme with cysteine and histidine residues located in its active site. Moreover, the enzyme exhibited remarkable stability towards sodium dodecyl sulfate (SDS) and Triton X-100. This is the first report of β-amylase production from moderate halophiles. The present study indicates that the extracellular β-amylase of Halobacillus sp. LY9 may have considerable potential for industrial application owing to its properties.
authors.affiliation: Life Science College, Yuncheng University, Yuncheng 044000, China. lixin-eva@163.com
authors.forename: Xin
authors.initials: X
authors.lastname: Li
authors.forename: Hui-Ying
authors.initials: HY
authors.lastname: Yu
mesh_terms: D002681:China; D058678:Halobacillus / Q000201:enzymology* / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D008320:Maltose / Q000378:metabolism; D012965:Sodium Chloride / Q000494:pharmacology; D013213:Starch / Q000378:metabolism; D013696:Temperature; D001614:beta-Amylase / Q000096:biosynthesis / Q000378:metabolism*
chemical_list: D012965:Sodium Chloride; D008320:Maltose; D013213:Starch; D001614:beta-Amylase