Isolation,Purification and Identification of Metalloprotease FQME14 from Fusarium sp. Q7-31T

doi:10.13560/j.cnki.biotech.bull.1985.2017-0896

Abstract

Abstract: The objectives of this work include：（1）to isolate the metalloproteinase from the fermented crude enzymatic solution of Fusarium sp. Q7-31T by oat straw,and then purify and identify them;（2）to study the enzymatic characteristics of protease for enriching and improving the basic data of metalloproteinases;and（3）to explore the role of cellulase in the degradation process of cellulose for understanding the mechanisms of cellulase degradation. Strain was cultured in liquid fermentation with oat straw as carbon source,and then crude enzyme solution was collected on the sixth day. The metalloprotease was purified using ammonium sulfate precipitation,Sephacry S-100 gel filtration chromatography and DEAE weak ion-exchange chromatography,its enzymatic properties were studied,and it was identified by MADIL-TOF-TOF. As results,the protease FQME14 isolated from the crude enzyme solution of Fusarium sp. Q7-31T was proved by SDS-PAGE to be a single band with a relative molecular mass of 30 kD. The MADIL-TOF-TOF results suggested that protease FQME14 belonged to M14 family. The characterization of purified protease showed that the specific activity of protease was 2.85 U/mg,its optimal pH and temperature for the enzyme activity were 7.0 and 40℃,respectively. The protease was very stable at pH 5.0-9.0 and temperature up to 50℃. The protease presented high degradation capacity to casein,ovalbumin and bovine serum albumin. The Co²⁺and Mn²⁺ were stimulators for protease,while Fe²⁺,Mg²⁺,K⁺,Ca²⁺,Zn²⁺and Na⁺ ions inhibited the activity of the protease at varied level. In conclusion,a protease from Fusarium sp. Q7-31T,named FQME14,was isolated,purified,identified and characterized. The enzymatic properties and MADIL-TOF-TOF results suggest that protease FQME14 belongs to M14 family,and is a new metalloprotease.

Key words: fusarium, metalloprotease, chromatography, identify, M14 family

QIN Ri-tian1LU Yu-li1LIANG Gao-li1XIE Zhan-ling1, 2LEI Ya-nan3. Isolation,Purification and Identification of Metalloprotease FQME14 from Fusarium sp. Q7-31T[J]. Biotechnology Bulletin, 2018, 34(4): 161-167.

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