کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10129180 | 1645202 | 2018 | 32 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Heterologous expression of an agarase gene in Bacillus subtilis, and characterization of the agarase
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موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
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چکیده انگلیسی
A β-agarase was identified from Pseudoalteromonas sp. Q30F and heterologously expressed in Bacillus subtilis WB800n. The β-agarase, Aga862 encoded by aga862 gene in an open reading frame of 1338â¯bp is 445 amino acids in length, and has a calculated molecular mass of 50.1â¯kDa and an estimated isoelectric point of 4.81. Protein sequence analysis showed that Aga862 belongs to family 16 of glycoside hydrolases (GH16) and carbohydrate-binding module family 13 (CBM13). The agarase was expressed in B. subtilis WB800n and purified by precipitation, anion exchange and gel filtration for a specific activity of 4.6â¯U/mg, a 27.8-fold improvement over the activity of the crude enzyme. Aga862 exhibited optimal activity at 45â¯Â°C and pHâ¯6.5, and showed excellent pH stability with retention of over 80% relative activities after preincubation for the pH range of 3.0-10.0 at 4â¯Â°C for 3â¯h. The agarase exhibited a Km value of 14.15â¯mg/mL toward agarose and a Vmax of 256.41â¯U/mg. The mass spectrometry analysis revealed that the end products of agar degradation were neoagarotetraose and neoagarohexaose. Recombinant Aga862 has great potential for the manufacture of agaro-oligosaccharides for the non-pathogenic nature and safety of the B. subtilis WB800n.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: International Journal of Biological Macromolecules - Volume 120, Part A, December 2018, Pages 657-664
Journal: International Journal of Biological Macromolecules - Volume 120, Part A, December 2018, Pages 657-664
نویسندگان
Xin Cui, Yuechen Jiang, Liuyi Chang, Lei Meng, Junhong Yu, Chun Wang, Xiaolu Jiang,