Heterologous production of a temperature and pH-stable laccase from Bacillus vallismortis fmb-103 in Escherichia coli and its application
| Autor: | Meixia Zheng, Jianna Sun, Chong Zhang, Fengxia Lu, Zhaoxin Lu |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0106 biological sciences
0301 basic medicine Ammonium sulfate Bioengineering medicine.disease_cause 01 natural sciences Applied Microbiology and Biotechnology Biochemistry law.invention 03 medical and health sciences chemistry.chemical_compound law 010608 biotechnology medicine Malachite green Escherichia coli Laccase biology biology.organism_classification Bacillus vallismortis 030104 developmental biology chemistry Yield (chemistry) Recombinant DNA Fermentation |
| Zdroj: | Process Biochemistry. 55:77-84 |
| ISSN: | 1359-5113 |
| DOI: | 10.1016/j.procbio.2017.01.030 |
| Popis: | To enhance laccase yield, the laccase gene from Bacillus vallismortis fmb-103 was cloned and heterologously expressed in Escherichia coli BL21 (DE3) cells. The auto-induction strategy was applied during fermentation, and the process was controlled, as follows: Cu2+ was added when the optical density at 600 nm (OD600) was 0.3, the fermentation temperature was adjusted to 16 °C when the OD600 was 0.9, and fermentation was stopped after 50 h. The yield of recombinant laccase was up to 3420 U/L, as assayed by 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid). Recombinant laccase was purified 4.47-fold by heating for 10 min at 70 °C and dialyzing against 50–60% ammonium sulfate, retained more than 50% activity after 10 h at 70 °C, and demonstrated broad pH stability. Malachite green was efficiently degraded by recombinant laccase, especially in combination with mediators. These results provided a basis for the future application of recombinant laccase to malachite green degradation. |
| Databáze: | OpenAIRE |
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