Expression of the antimicrobial immunopeptide Scy based on the engineered bacteria EcN and its application

ZHOU Lutong, JIANG Xiao, JIN Mingliang, WANG Yizhen, CHENG Yuanzhi

PDF(778 KB)
PDF(778 KB)
Animals Breeding and Feed ›› 2025, Vol. 24 ›› Issue (03) : 1-7. DOI: 10.13300/j.cnki.cn42-1648/s.2025.03.001

Expression of the antimicrobial immunopeptide Scy based on the engineered bacteria EcN and its application

Author information +
History +

Abstract

Objectives Escherichia coli Nissle 1917 (EcN) was used to express antimicrobial immune peptide Scygonadin (Scy) and the effects of EcN-Scy on alleviating colitis were evaluated to provide theoretical basis for the development of novel substitutes for antibiotic. Methods The vector pnirBMisl (pnir) and the target gene of antimicrobial peptide Scy were amplified with PCR, and the recombinant vector EcN-Scy was obtained by homologous recombination connection. Twenty 8-week-old female BALB/c mice were randomly divided into four groups including the control group, DSS group, DSS+EcN group, and DSS+EcN Scy group. The mice were orally administered with PBS, EcN bacterial solution, or EcN-Scy bacterial solution every 2 days and their body weight was recorded. The experiment lasted for 14 days. Except for the control group, the other three groups were induced colitis by adding 3% DSS to their drinking water from day 0 to day 7. Mice were killed on day 14 after anesthesia, and samples of serum and colon tissue were collected for testing. Results EcN-Scy treatment significantly alleviated the weight loss in mice (P<0.05) and the symptoms of colon shortening caused by DSS (P<0.05), and significantly improved the structure of colonic villi compared with DSS group. The level of ALP and Urea in serum in the DSS+EcN-Scy group was significantly increased by 28.6 U/L and 1.35 mol/L (P<0.05) compared with that in DSS group. The level of TBil and LDH had a decreasing trend (P>0.05). The level of pro-inflammatory cytokines TNF-α, MCP1, and IL-1 β in the serum of the DSS+EcN-Scy group was significantly reduced by 18.9%, 50.9%, and 8.9%, respectively (P<0.05), and the level of MCP1 and IL-6 in the colon were reduced by 40.4% and 21.9%, respectively (P<0.05) compared with that in the DSS group. Conclusions The antimicrobial immune peptide Scy was successfully expressed with the engineered bacterium EcN, which has a significant effect on alleviating colitis in mice. It can improve the weight loss, colon shortening, and morphological damages in mice induced by DSS, and reduce the level of inflammatory factors in serum and colon.

Key words

Escherichia coli Nissle 1917 (EcN) / antimicrobial immune peptide Scygonadin (Scy) / recombinant expression / colitis / research and development of substitutes for antibiotic

CLC number

S816

Cite this article

Download Citations
ZHOU Lutong , JIANG Xiao , JIN Mingliang , et al . Expression of the antimicrobial immunopeptide Scy based on the engineered bacteria EcN and its application. Animals Breeding and Feed. 2025, 24(03): 1-7 https://doi.org/10.13300/j.cnki.cn42-1648/s.2025.03.001

References

1
OLDER C E, GRIFFIN M J, RICHARDSON B M, et al. Influence of probiotic and prebiotic supplementation on intestinal microbiota and resistance to Edwardsiella ictaluri infection in channel catfish ( Ictalurus punctatus ) following florfenicol administration[J/OL]. Journal of fish diseases, 2024, 47(4): e13910[2024-11-28].
2
周雪莼,胡婷婷,王佳慧,等.高效液相色谱-高分辨质谱法快速筛查动物源性药食同源产品中32种抗生素兽药残留[J].吉林中医药,2023,43(12): 1469-1474.
3
CARDOSO O, ASSIS G, DONATO M M, et al. Antibiotic residues and zinc concentrations in the livers and kidneys of portuguese piglets: relationship to antibiotic and zinc resistance in intestinal Scherichia coli [J]. Biological trace element research, 2024, 202(10): 4522-4530.
4
XU B, FU J, ZHU L, et al. Overall assessment of antibiotic substitutes for pigs: a set of meta-analyses[J/OL]. Journal of animal science and biotechnology, 2021, 12(1): 3[2024-11-27].
5
韩菲菲,安沙,谢永刚,等.猪乳铁蛋白肽的分子改良及改良肽的生物学活性研究[J].动物营养学报, 2011, 23(2): 241-249.
6
WEN C, ZHANG H, GUO Q, et al. Engineered Bacillus subtilis alleviates intestinal oxidative injury through Nrf2-Keap1 pathway in enterotoxigenic Escherichia coli (ETEC) K88-infected piglet[J]. Journal of Zhejiang University-SCIENCE B, 2023, 24(6): 496-509.
7
XU B, WANG L, YANG C, et al. Specifically targeted antimicrobial peptides synergize with bacterial-entrapping peptide against systemic MRSA infections[J/OL]. Journal of advanced research, 2024: S2090123224000365[2024-11-28].
8
ZONG X, CAO X, WANG H, et al. Porcine lactoferrin-derived peptide LFP-20 modulates immune homoeostasis to defend lipopolysaccharide-triggered intestinal inflammation in mice[J]. British journal of nutrition, 2019, 121(11): 1255-1263.
9
YANG E, SHEN J. The roles and functions of Paneth cells in Crohn’s disease: a critical review[J/OL]. Cell proliferation, 2021, 54(1): e12958[2024-11-28].
10
CHAUHAN P, TAPWAL A. Green synthesis of nanoparticles using botanicals and their application in management of fungal phytopathogens: a review[J/OL]. Archives of microbiology, 2023, 205(3): 94[2024-11-28].
11
MIYASHITA M, SAKAI S, OKABE R, et al. Identification and synthesis of a long‐chain antimicrobial peptide from the venom of the Liocheles australasiae scorpion[J/OL]. Journal of peptide science, 2024: e3661[2024-11-28].
12
苏琰, 李融.抗菌肽的食品保鲜应用及生物合成研究进展[J].食品与机械, 2024,40(7): 208-215.
13
罗会颖, 姚斌. 理性调控甲醇定向生物合成酵母单细胞蛋白的新策略[J]. 中国科学:生命科学, 2024,54(8): 1488-1490.
14
李朔, 李潇, 张晓黎, 等. 乳酸菌产γ-氨基丁酸及生物合成技术研究进展[J]. 中国调味品, 2022,47(11): 205-210.
15
沈晓熳, 杨炽光, 翁晨虹, 等.枯草芽孢杆菌RN8的抗菌活性成分分离及其作用机制[J].中国生物工程杂志, 2024,44(7): 12-28.
16
JIANG R, YUAN S, ZHOU Y, et al. Strategies to overcome the challenges of low or no expression of heterologous proteins in Escherichia coli [J/OL]. Biotechnology advances, 2024, 75: 108417[2024-11-28].
17
郭子好, 方华, 李春源, 等. 锯缘青蟹Scygonadin抗菌免疫肽的毕赤酵母表达及其产物抑菌活性分析[J].饲料研究, 2017(10): 32-36.
18
KAI L, ZONG X, JIANG Q, et al. Protective effects of polysaccharides from Atractylodes macrocephalae Koidz. against dextran sulfate sodium induced intestinal mucosal injury on mice[J]. International journal of biological macromolecules, 2022, 195: 142-151.
19
田家华, 索小涛, 马晨曦, 等. 溃疡性结肠炎动物模型构建方法的研究进展[J]. 山东医药, 2024,64(13): 111-114.
20
高梦学, 王丽娜, 黄鹤. 合成生物学在肠道微生态疗法研发中的应用[J]. 合成生物学, 2022, 3(1): 35-52.
21
卢雯, 刘睿娜, 雷璞, 等.基因工程菌的构建、性能研究及其在疾病诊断和治疗中的应用[J].微生物学报, 2023,63(10): 3758-3772.
22
XIE W, SONG L, WANG X, et al. A bovine lactoferricin-lactoferrampin-encoding Lactobacillus reuteri CO21 regulates the intestinal mucosal immunity and enhances the protection of piglets against enterotoxigenic Escherichia coli K88 challenge[J/OL]. Gut Microbes, 2021, 13(1): 1956281[2024-11-28].

Comments

PDF(778 KB)

Accesses

Citation

Detail

Sections
Recommended

/