Optimization, Characteristics, and Functions of Alkaline Phosphatase From Escherichia coli

Yachao Dong; Yandong Xia; Jie Yin; Diao Zhou; Yidan Sang; Sufeng Yan; Qingshu Liu; Yaqi Li; Leli Wang; Ying Zhao; Cang Chen; Qiuyun Huang; Ying Wang; Muhammad Nazeer Abbasi; Huansheng Yang; Chuni Wang; Jianzhong Li; Qiang Tu; Jia Yin

doi:10.3389/fmicb.2021.761189

Optimization, Characteristics, and Functions of Alkaline Phosphatase From Escherichia coli

Front Microbiol. 2022 Feb 21:12:761189. doi: 10.3389/fmicb.2021.761189. eCollection 2021.

Authors

Yachao Dong^{1

2}, Yandong Xia³, Jie Yin¹, Diao Zhou¹, Yidan Sang¹, Sufeng Yan¹, Qingshu Liu⁴, Yaqi Li¹, Leli Wang¹, Ying Zhao¹, Cang Chen¹, Qiuyun Huang¹, Ying Wang¹, Muhammad Nazeer Abbasi², Huansheng Yang¹, Chuni Wang¹, Jianzhong Li¹, Qiang Tu², Jia Yin¹

Affiliations

¹ Hunan Provincial Key Laboratory of Animal Intestinal Function and Regulation, Hunan International Joint Laboratory of Animal Intestinal Ecology and Health, The National and Local Joint Engineering Laboratory of Animal Peptide Drug Development, College of Life Sciences, Hunan Normal University, Changsha, China.
² CAS Key Laboratory of Quantitative Engineering Biology, Shenzhen Institute of Synthetic Biology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences (CAS), Shenzhen, China.
³ College of Life Sciences and Technology, Central South University of Forestry and Technology, Changsha, China.
⁴ Hunan Institute of Microbiology, Changsha, China.

Abstract

Weaning of piglets could increase the risk of infecting with Gram-negative pathogens, which can further bring about a wide array of virulence factors including the endotoxin lipopolysaccharide (LPS). It is in common practice that the use of antibiotics has been restricted in animal husbandry. Alkaline phosphatase (AKP) plays an important role in the detoxification and anti-inflammatory effects of LPS. This study investigated the protective effects of AKP on intestinal epithelial cells during inflammation. Site-directed mutagenesis was performed to modulate the AKP activity. The enzyme activity tests showed that the activity of the DelSigD₁₅₃G-D₃₃₀N mutants in B. subtilis was nearly 1,600 times higher than that of the wild-type AKP. In this study, an in vitro LPS-induced inflammation model using IPEC-J2 cells was established. The mRNA expression of interleukin-(IL-) 6, IL-8, and tumor necrosis factor-α (TNF-α) were extremely significantly downregulated, and that of ASC amino acid transporter 2 (ASCT-2), zonula occludens protein-1 (ZO-1), and occludin-3 (CLDN-3) were significantly upregulated by the DelSigD₁₅₃G-D₃₃₀N mutant compared with LPS treatment. This concludes the anti-inflammatory role of AKP on epithelial membrane, and we are hopeful that this research could achieve a sustainable development for the pig industry.

Keywords: IPEC-J2 cell line; alkaline phosphatase; disease; immunity; inflammatory factor.