The purpose of this study was to solve the problem that glucose oxidase (GOD) from marine bacteria has low activity. The genomic DNA of Citrobacters sp. 8-III was extracted and purified, and the target sequence was obtained by comparing against existing GOD genes. The synthetic and codon-optimized GOD gene was subcloned into the vector pET28a(+), and the recombinant expression vector pET28a(+)-GOD was constructed and transformed into Escherichia coli BL21(DE3) for expression. The recombinant GOD was purified by nickel affinity chromatography, and its enzymatic properties were studied. It was found that E. coli BL21(DE3)/pET-28a(+)-GOD producing GOD was successfully constructed. The purified GOD was 46 kDa. The activity of recombinant GOD was 2.04 U/mL, and its optimal reaction temperature and pH were 25 ℃ and 6.0, respectively. Additionally, K+ and Ni2+ significantly promoted its activity. Moreover, recombinant GOD could significantly accelerate the growth of chicken and it had good antiseptic effects in feeds of puppies. In conclusion, this study firstly introduced the marine bacterial GOD gene into E. coli to reveal a new host of GOD. Besides, recombinant GOD has cold-active enzyme properties, which lays a foundation for its applications in feed additives and cryogenics.
LIU Chunying
,
HU Shansong
,
ZHANG Qingfang
,
LI Meiyu
,
YU Shuang
,
CHI Naiyu
. Cloning and expressing low temperature glucose oxidase gene from marinebacteria in Escherichia coli[J]. Food and Fermentation Industries, 2019
, 45(11)
: 34
-39
.
DOI: 10.13995/j.cnki.11-1802/ts.019994
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