Antibiofilm activity and synergistic effects of DNase I and lysostaphin against Staphylococcus aureus biofilms

Author:

Lin Quan123,Sheng Maokun123,Tian Yanjun4,Li Bing5,Kang Zhaodi123,Yang Yingying123,Xu Zhenbo678,Soteyome Thanapop8,Guo Liang9,Sun Huzhi10,Gao Yan11,Yu Lingyun12,Pan Qiang1234,Tan Yulong123

Affiliation:

1. Special Food Research Institute, Qingdao Agricultural University , Qingdao , China

2. Shandong Technology Innovation Center of Special Food , Qingdao , China

3. Qingdao Special Food Research Institute , Qingdao , China

4. Qingdao NABT PhagePharm Co., Ltd. , Qingdao , China

5. Marine Science and Engineering College, Qingdao Agricultural University , Qingdao , China

6. School of Food Science and Engineering, Guangdong Province Key Laboratory for Green Processing of Natural Products and Product Safety, Engineering Research Center of Starch and Vegetable Protein Processing Ministry of Education, South China University of Technology , Guangzhou , China

7. Department of Biology, Massachusetts Institute of Technology , Cambridge, MA , USA

8. Home Economics Technology, Rajamangala University of Technology Phra Nakhon , Bangkok , Thailand

9. China animal husbandry industry Co., Ltd. , Beijing , China

10. Key Lab of Preventive Veterinary Medicine in Hubei Province, College of Veterinary Medicine, Huazhong Agricultural University , Wuhan , China

11. Marine Science Research Institute of Shandong Provice (National Oceanographic Center of Qingdao) , Qingdao , China

12. Qingdao PhagePharm bio-tech Co., Ltd. , Qingdao , China

Abstract

Abstract Background In recent years, food safety has become a global public health concern. Microbial contamination is one of the most common food safety issues. Staphylococcus aureus is a common foodborne pathogen that can form biofilms on the surface of food processing equipment, leading to greater resistance to antimicrobial agents than occurs with planktonic bacteria. Materials and methods In this work, recombinant Escherichia coli BL21 (DE3) cells expressing optimized lysostaphin (Lst) were constructed, recombinant Lst was produced and purified, and Lst enzymatic assays were performed, followed by antimicrobial testing of Lst. Finally, a mixture of Lst and DNase I was tested for antibiofilm activity. Results The protein content of purified Lst was 0.6 mg/mL and the enzyme activity was 240 U/mL. The minimum inhibitory concentration (MIC) of Lst against S. aureus was 0.1 μg/mL. At 1 MIC, Lst exerted an effect on the growth, cell wall integrity and cell membrane permeability of S. aureus. Conclusions Although Lst alone also showed good inhibition and disruption of S. aureus biofilms, the inhibition and disruption of S. aureus biofilms were significantly greater when Lst was mixed with DNase I. This is probably because DNase I removes extracellular DNA, affecting biofilm formation and dispersing mature biofilms, and thereby facilitating the penetration of Lst.

Funder

Natural Science Foundation of Shandong Province

Young Taishan Scholars Program of Shandong Province

Talent Research Foundation of Qingdao Agricultural University

Research Fund of Qingdao Special Food Research Institute

Publisher

Oxford University Press (OUP)

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