Optimization of purification protocol of biomimetic affinity material for marine metalloprotease MP

YANG Juan; LI Shang-yong; MA Zi-bin; LIU Jun-zhong; HAO Jian-hua; WANG Yue-jun; SUN Mi

doi:10.13386/j.issn1002-0306.2017.15.028

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Science and Technology of Food Industry > 2017 > (15): 146-152. > DOI: 10.13386/j.issn1002-0306.2017.15.028

YANG Juan, LI Shang-yong, MA Zi-bin, LIU Jun-zhong, HAO Jian-hua, WANG Yue-jun, SUN Mi. Optimization of purification protocol of biomimetic affinity material for marine metalloprotease MP[J]. Science and Technology of Food Industry, 2017, (15): 146-152. DOI: 10.13386/j.issn1002-0306.2017.15.028

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Optimization of purification protocol of biomimetic affinity material for marine metalloprotease MP

Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Laboratory for Marine Drugs and Bioproducts of Qingdao National Laboratory for Marine Science and Technology
College of Food Sciences&Technology, Shanghai Ocean University

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Received Date: February 20, 2017

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Abstract

Objective: To establish an efficient method for biomimetic affinity purification of marine metalloprotease MP.Methods: Using the biomimetic affinity materials to optimize the purification conditions by response surface optimization method, such as the single factor experiment, Plackett-Burman experiment design, Box-Behnken experiment.And the purity of purified samples by SDS-PAGE and HPLC was analyzed. Results: The purification conditions were optimized by single factor experiment and the response surface design. The study indicated that the best purification conditions of the boronic acid gel were: When enzyme concentration was 100 mg/m L, the optimal loading buffer was glycine-sodium hydroxide of p H8.6 and the sample rate was 1 m L/min. The elution buffer was disodium hydrogen phosphate-citric acid of p H5.2, and elution rate was2 m L/min, the elution buffer added to mannitol of 75 mmol/L, the purification efficiency was enhanced 11 times and the purity above 98.8%.
- Metalloproteinase MP,
- affinity chromatography,
- purification

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Optimization of purification protocol of biomimetic affinity material for marine metalloprotease MP