Bacterial Expression and Characterization of Recombinant β-Xylosidase from the Thermophilic Xylanolytic Bacterium Bacillus sp

Background: Amylases are used in several industrial and biotechnological sectors, including those producing textiles, detergents, paper and bakery products. Objective: This study aimed to purify an industrially important α-amylase from Bacillus sp. For this purpose, a single and rapid α-amylase purification was performed using the starch affinity method. Methods: Characterization of the purified enzyme was determined by investigating temperature, pH stability, detergents, and metal ions. Results: The purification coefficient of 29.8-fold with a yield of 9.2% was found. The molecular weight of the purified α-amylase was determined to be 53 kDa by SDS-PAGE, and thermostability was confirmed with 100% activity at 30ºC and 40ºC after 1 h. The purified enzyme was stable over a wide range of pH values, with optimum activity at pH 6.0, 7.0 and 8.0 after 2 h. The study also investigated the effects of the metal ions and detergents on the purified amylase and found that Mg2+ and Ca2+ ions were the activators of the enzyme, while Zn2+, Co2+ and Na+ ions decreased the activity. Furthermore, Hg2+ indicated complete inhibition of amylase activity. The detergents Triton X-100 and Tween 20 increased the α-amylase activity, while sodium dodecyl sulfate inhibited the activity. Conclusion: The purified α-amylase obtained from Bacillus sp. is considered to be environmentally friendly, can be processed in a short time, and has a low cost.

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Purification and characterization of an alkaline chloride-tolerant laccase from a halotolerant bacterium, Bacillus sp. strain WT

Journal of Molecular Catalysis B Enzymatic ◽

10.1016/j.molcatb.2016.10.001 ◽

2016 ◽

Vol 134 ◽

pp. 89-97 ◽

Cited By ~ 29

Author(s):

Maryam Siroosi ◽

Mohammad Ali Amoozegar ◽

Khosro Khajeh

Keyword(s):

Bacillus Sp ◽

Purification And Characterization ◽

Halotolerant Bacterium ◽

Bacterium Bacillus

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Anaerobic synthesis of extracellular proteases by the soil bacterium Bacillus sp. AM-23: Purification and characterization of the enzymes

Soil Biology and Biochemistry ◽

10.1016/0038-0717(95)00094-u ◽

1995 ◽

Vol 27 (11) ◽

pp. 1377-1382 ◽

Cited By ~ 6

Author(s):

Kenji Aoki ◽

Kukizo Miyamoto ◽

Shuichiro Murakami ◽

Ryu Shinke

Keyword(s):

Soil Bacterium ◽

Bacillus Sp ◽

Extracellular Proteases ◽

Purification And Characterization ◽

Bacterium Bacillus

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Cloning and Characterization of a Cold-adapted Chitosanase from Marine Bacterium Bacillus sp. BY01

Molecules ◽

10.3390/molecules24213915 ◽

2019 ◽

Vol 24 (21) ◽

pp. 3915 ◽

Cited By ~ 3

Author(s):

Yue Yang ◽

Zhou Zheng ◽

Yifei Xiao ◽

Jiaojiao Zhang ◽

Yu Zhou ◽

...

Keyword(s):

Glycoside Hydrolase ◽

Marine Bacterium ◽

Volume Concentration ◽

Biological Activities ◽

Maximum Activity ◽

Bacillus Sp ◽

Cold Adapted ◽

Encoding Gene ◽

Bacterium Bacillus

Chitosanase plays an important role in the production of chitooligosaccharides (CHOS), which possess various biological activities. Herein, a glycoside hydrolase (GH) family 46 chitosanase-encoding gene, csnB, was cloned from marine bacterium Bacillus sp. BY01 and heterologously expressed in Escherichia coli. The recombinant chitosanase, CsnB, was optimally active at 35 °C and pH 5.0. It was also revealed to be a cold-adapted enzyme, maintaining 39.5% and 40.4% of its maximum activity at 0 and 10 °C, respectively. Meanwhile, CsnB showed wide pH-stability within the range of pH 3.0 to 7.0. Then, an improved reaction condition was built to enhance its thermostability with a final glycerol volume concentration of 20%. Moreover, CsnB was determined to be an endo-type chitosanase, yielding chitosan disaccharides and trisaccharides as the main products. Overall, CsnB provides a new choice for enzymatic CHOS production.

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