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Cloning, Heterologous Expression, and Characterization of a Neutral Uricase from Arthrobacter sp. CSAJ-16 in Cangshan Mountain

Xin Yan
Xin Yan
, 
Wei Hu
Wei Hu
, 
Yun-Guo Zhu
Yun-Guo Zhu
, 
Qing-Qing Liu
Qing-Qing Liu
, 
Shuai Wang
Shuai Wang
, 
Hong-Yan Liu
Hong-Yan Liu
, 
Dan Zhu
Dan Zhu
, 
Zhi-Hua Lv
Zhi-Hua Lv
, 
Lin-Hua Li
Lin-Hua Li
 and 
Yi-Rui Yin
Yi-Rui Yin
   | Sep 20, 2023
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Article Category: Original Paper
Published Online: Sep 20, 2023
Page range: 277 - 283
Received: Apr 10, 2023
Accepted: Jun 09, 2023
DOI: https://doi.org/10.33073/pjm-2023-027
Keywords
© 2023 Xin Yan et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig. 1.

Growth and uric acid degradation curve of Arthrobacter sp. CSAJ-16.
Growth and uric acid degradation curve of Arthrobacter sp. CSAJ-16.

Fig. 2.

The phylogenetic tree of Arthrobacter sp. CSAJ-16 base on the 16S rRNA sequences.
The phylogenetic tree of Arthrobacter sp. CSAJ-16 base on the 16S rRNA sequences.

Fig. 3.

SDS-PAGE analysis of the expression of ArUOX in the recombinant strain UOX-CSAJ-16.M – Molecular weight standard, Lane 1 – crude enzyme, Lane 2 – purified ArUOX
SDS-PAGE analysis of the expression of ArUOX in the recombinant strain UOX-CSAJ-16.M – Molecular weight standard, Lane 1 – crude enzyme, Lane 2 – purified ArUOX

Fig. 4.

Effects of temperature (A) and pH (B) on the activity of ArUOX.
Effects of temperature (A) and pH (B) on the activity of ArUOX.

Fig. 5.

Effects of temperature (A) and pH (B) on the stability of ArUOX.
Effects of temperature (A) and pH (B) on the stability of ArUOX.

PCR primers. Underlined sequences represent the homologous recombinant fragment with the pET28a(+).

Primer type Primer name Sequences (5′–3′)
Universal primers 27F AGAGTTTGATCCTGGCTCAG
1492R GGTTACCTTGTTACGACTT
Amplification primersI 1-Arthrobacter saudimassiliensis-Uox-F ATGACCGCAACAGAACAGGCG
1-Arthrobacter saudimassiliensis-Uox-R TCAGCAGAACCCCGCGATGGT
2-Arthrobacter globiformis-Uox-F ATGAGCAACAAGATCGTCCTCG
2-Arthrobacter globiformis-Uox-R CTAGCAGAAGCCGGCGATGC
3-Arthrobacter liuii-Uox-F ATGAGCAGCAAGATCATCCT
3-Arthrobacter liuii-Uox-R TTAGCAGAAGCCGGCGATGC
4-Arthrobacter ulcerisalmonis-Uox-F ATGAGCAGCAAGATCATCCTC
4-Arthrobacter ulcerisalmonis-Uox-R TCAGCAGAATCCGGCGATGC
5-Arthrobacter rhombi-Uox-F ATGACTACCACCGCTTCTCAG
5-Arthrobacter rhombi-Uox-R TTAGCAGAAGCCTGCGGCAT
6-Arthrobacter crystallopoietes-Uox-F ATGACCGCCACCGTGGAATC
6-Arthrobacter crystallopoietes-Uox-R GCAGAATCCGGGGATATTTG
7-Arthrobacter arilaitensis-Uox-F ATGACTATCGAAACCACCGCC
7-Arthrobacter arilaitensis-Uox-R GGCAAATGCCGGGATATTGGA
8-Arthrobacter dokdonellae-Uox-F ATCATCCTGGGCGCCAACC
8-Arthrobacter dokdonellae-Uox-R GCAGAAGCCTGCGACGCCG
Amplification primersII TFH-3- Arthrobacter liuii-Uox-F CAAATGGGTCGCGGATCCGAAATGAGCAGCAAGATCATCCT
TFH-3- Arthrobacter liuii-Uox-R GTGCTCGAGTGCGGCCGCAAGTTAGCAGAAGCCGGCGATGC

Effect of different ions and chemical reagents on enzyme activity.

Chemicals Final concentration Residual activity (%)
Blank 10 mM 100.0
K+ 10 mM 250.72 ± 2.90
Mg2+ 10 mM 279.71 ± 3.34
Fe2+ 10 mM 43.48 ± 0.35
Ca2+ 10 mM 204.35 ± 4.35
Ni2+ 10 mM 181.16 ± 2.90
Co2+ 10 mM 17.39 ± 0.33
Ba2+ 10 mM 256.52 ± 2.45
Mn2+ 10 mM 23.19 ± 0.65
Pb2+ 10 mM 256.52 ± 4.35
Zn2+ 10 mM 0
EDTA 10% 26.09 ± 0.34
SDS 10% 0
PSMF 10% 53.62 ± 0.79
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