alexa Efficient Isolation and Functional Analysis of Spontaneous Streptococcus thermophilus Bacteriophage-Insensitive Mutants
E- ISSN: 2320 - 3528
P- ISSN: 2347 - 2286

Research & Reviews: Journal of Microbiology and Biotechnology
Open Access

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Research Article

Efficient Isolation and Functional Analysis of Spontaneous Streptococcus thermophilus Bacteriophage-Insensitive Mutants

Chengjie Ma1,2, Jinzhong Lin1,2, Guangyu Gong1,2, Zhenmin Liu1,2, Zhengjun Chen3* and Aimin Ma4*

1State Key Laboratory of Dairy Biotechnology, Technology Center of Bright Dairy & Food Co. Ltd., Shanghai 200436, PR China

2Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University, Wuxi 214122, PR China

3State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, PR China

4College of Food Science and Technology, Huazhong Agricultural University, No.1 Shizishan Street, Wuhan 430070, PR China

*Corresponding Author:
Zhengjun Chen
College of Life Science and Technology, Huazhong Agricultural University, No.1 Shizishan Street, Wuhan 430070, PR China
Tel: 86-27-87281267
Fax: 86-27-87280670
E-mail: [email protected]

Aimin Ma
College of Food Science and Technology, Huazhong Agricultural University, No.1 Shizishan Street, Wuhan 430070, PR China
Tel: +86278728211
Fax: +862787396057
E-mail: [email protected]

Received date: 31/08/2015 Accepted date: 25/02/2016 Published date: 10/03/2016

 

Abstract

The use of bacteriophage-resistant strains that have satisfactory technological properties is essential for preventing phage infections in industrial fermentation processes. Here, we describe an improved method for the isolation of spontaneous Streptococcus thermophilus bacteriophage-insensitive mutants (BIMs). The concentration of large volumes of S. thermophilus secondary cultures, their subculture in skim milk with high titers of bacteriophages, and their inoculation with the fast milk acidifying phenotype over multiple passages were critical to increasing the probability of obtaining mutants, improving isolation efficiency, and maintaining technological performance. We obtained large quantities of BIMs after every round of screening, with isolation efficiency in excess of 85%. We analyzed nine BIMs, and these were all similar to their parent strain S. thermophilus St1 with respect to syneresis, water holding capacity, and apparent viscosity. In comparison to the parent strain, seven of the nine variants possessed equivalent acidifying activities, with the remaining two variants exhibiting excellent acidification performance. One or two new spacers from corresponding phages were found in all nine BIMs, demonstrating that the CRISPR/Cas system was responsible for the phage resistance of S. thermophilus.

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