FOOD SCIENCE ›› 2017, Vol. 38 ›› Issue (12): 104-111.doi: 10.7506/spkx1002-6630-201712016

• Bioengineering • Previous Articles     Next Articles

Isolation, Purification and Characterization of a Novel Carotenoid-Cleaving Enzyme from Staphylococcus pasteuri TS-82

ZHU Mingming, FAN Mingtao, HE Hongju, MA Hanjun   

  1. 1. School of Food Science, Henan Institute of Science and Technology, Xinxiang 453003, China; 2. College of Food Science and Engineering, Northwest A&F University, Yangling 712100, China
  • Online:2017-06-25 Published:2017-06-26

Abstract: A carotenoid cleavage enzyme with HPLC grade purity of 95.6% was obtained from Staphylococcus pasteuri (S. pasteuri) TS-82 by using anion-exchange chromatography, preparative high performance liquid chromatography (PHPLC), and superdex peptide chromatography together. The purified enzyme had a specific activity of 125 U/g with 466-fold purification and 2.39% recovery. Its molecular weight was 655.093 D as identified by liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI/MS). Enzymatic characterization indicated that the purified enzyme had optimal temperature of 60 ℃ for degrading C40 carotenoids and 50 ℃ for degrading β-apo-8?ˉ-carotenal, and it maintained stable activity at 50 ℃. The optimum pH value was 3.0 for degrading all investigated substrates. The Km and vmax values indicated that the enzyme showed the highest affinity towards zeaxanthin, followed by astaxanthin, β-carotene, canthaxanthin, and β-apo-8?ˉ-carotenal in a decreasing order. The metal ions Al3+ and Fe3+ were identified as potent activators for the purified enzyme, whereas Fe2+ as a potent inhibitor. H2O2 was able to increase the enzyme activity in degrading β-carotene at levels of 0-16 mmol/L. Alcohol at low concentration (4%–16%) did not inhibit the activity of the enzyme. In conclusion, the enzyme showed great acid resistance and thermostability, which makes it stable in wine environment and provides a basis for industrial application.

Key words: Staphylococcus pasteuri TS-82, carotenoid cleavage enzyme, carotenoids, purification, characterization

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