Identification and Analysis of a Nattokinase-producing Strain

doi:10.13560/j.cnki.biotech.bull.1985.2016.01.030

Abstract

Abstract: The objective of this work is to identify an isolate Td of yielding high nattokinase, and to analyze the molecular characteristics of its nattokinase. The morphologic, biochemical and physiological characterizations, and molecular biological methods were used to identify the taxonomic position of isolate Td;mass spectrometric determination and analysis of MALDI-TOF, SDS-PAGE and fibrinolytic activity were employed to measure the characteristics of nattokinase, and PCR method was adopted to clone the full length of nattokinase gene. The bacterial strain Td was identified as Bacillus subtilis subsp. subtilis according to the results of morphologic, biochemical and physiological characterizations, as well as 16S rDNA, the sequences of 16S rDNA gyrA, hybrid rate of DNA-DNA. The yield of nattokinase produced by strain Td reached 300 mg/L, accounted for over 40% of the total protein. Fibrinolytic activity was over 230 U/mL. Amino acid sequence of nattokinase produced by Td showed the highest similarity to the subtilisin E. and the full length of the gene was 1143 bp. Conclusively, as a high-yield and high-activity strain for nattokinase, B. subtilis subsp. subtilis Td is a promising candidate for industrial development and utilization.

Key words: Bacillus subtilis subsp. subtilis, nattokinase, strain identification

WANG Xue-yan, SUN Yu-fei, FENG Fei, CHEN Xiao-fei, LI Feng, ZHOU Fu-zhong. Identification and Analysis of a Nattokinase-producing Strain[J]. Biotechnology Bulletin, 2016, 32(1): 187-194.

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