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Published on: February 16, 2018
A novel heat-resistant hydroxycinnamic acid decarboxylase from Wickerhamomyces anomalus
Wei Yang1, Hexia Ding2, Maodie Zeng1
1Key Laboratory of Plant Resource Conservation and Germplasm Innovation in Mountainous Region (Ministry of Education), School of Liquor and Food Engineering, Guizhou University, Guiyang, 550025, Guizhou Province, China; Guizhou Province Key Laboratory of Fermentation Engineering and Biopharmacy, School of Liquor and Food Engineering, Guizhou University, Guiyang, 550025, China.
None:
Hydroxycinnamate decarboxylase (HCDC) can decarboxylate Hydroxycinnamic acid to produce vinylphenol. In this study, a new heat-resistant HCDC (WaHCDC) was obtained from Wickerhamomyces anomalus WA64 isolated from Daqu, the fermentation starter of Chinese Baijiu. Subsequently, WaHCDC was expressed in P. pastoris GS115. After purification with Ni2+-NTA resin, WaHCDC was obtained at 25 mg·L-1 with a molecular weight of 45 kDa. The enzyme activity of WaHCDC was 12.69 ± 0.07 U·mg-1. The relative activity of WaHCDC was still retained 57 % after incubating at 80 °C for 90 min. To enhance acid resistance, WaHCDC was immobilized on covalent organic frameworks (COFs) via in-situ encapsulation, achieving 80 % loading efficiency and a capacity of 1.5 mg·g-1. The immobilized activity was 14.35 ± 0.11 U·mg-1. The stability of the immobilized WaHCDC was improved at pH 3-6. Its conversion rate of ferulic acid (FA) to 4-vinylguaiacol was 13 % higher than that of the free WaHCDC after incubating at 40 °C for 120 min. The activity of immobilized WaHCDC was above 60 % after 7 cycles reusing. This study provides a new heat-resistant enzyme resource and efficient immobilization strategy for the green biosynthesis of vinylphenol.
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The reaction begins with transferring a proton from the acid catalyst to one of the hydroxyl groups, producing an oxonium ion.

