Isolation and identification of high-yielding alkaline phosphatase strain: a novel mutagenesis technique and optimization of fermentation conditions

Abstract

Isolating and screening enzyme-producing strains from microorganisms and the commercial production of ALPs from microorganisms are of increasing interest. In this work, isolation and identification of high-yielding alkaline phosphatase strain were carried out using atmospheric and room temperature plasma mutagenesis (ARTP) for optimization of fermentation conditions. A strain of alkaline phosphatase-producing bacteria was screened from soil and identified by 16S rRNA gene sequencing as Bacillus amyloliquefaciens and named S-1. This strain had an alkaline phosphatase activity of 2594.73 U/L. Later, mutagenesis breeding of the alkaline phosphatase-producing S-1 strain was conducted using (ARTP), from which a higher alkaline phosphatase-producing positive mutant strain S-52 was screened. A central combination of five factors, including corn starch, yeast extract, metal ions, fermentation temperature and inoculum ratio, was then used to influence the activity of alkaline phosphatase. Results from the response surface methodology showed that the maximum enzyme activity of alkaline phosphatase was 12,110.6 U/L at corn starch, yeast extract and magnesium ions concentrations of 17.48 g/L, 18.052 g/L and 0.744 g/L, respectively; fermentation temperature of 37.192 °C; and inoculation ratio of 5.59%. This study is important for further exploring ARTP mutagenesis in B. amyloliquefaciens and the commercialization of ALPs.

Keywords:

• Alkaline phosphatase
• atmospheric and room temperature plasma
• Bacillus amyloliquefaciens
• response surface methodology

Author contributions

Le Bo: Research, writing, software, validation, and revision. Xin Kang: Research, writing, and revision. Zuohui Chen: Research, writing, and revision. Yue Zhao: Research, writing, and revision. Wenxue Hui: Research, writing, and revision. Si Wu: Research, writing, and revision. Shuang Bao: Took the lead in organizing the collaboration, research, writing, and revision. Jie Li: Took the lead in organizing the collaboration, research, writing, and revision.

Disclosure statement

The authors declare that they have no known competing financial interests or personal relationships that could have influenced the work reported herein.

Additional information

Funding

The author(s) reported there is no funding associated with the work featured in this article.