TY - JOUR

T1 - Alternative Routes for the Production of Natural 4-Vinylguaiacol from Sugar Beet Fiber Using Basidiomycetous Enzymes

AU - Günther, Thorben

AU - Schoppe, Lasse

AU - Ersoy, Franziska

AU - Berger, Ralf G.

N1 - Funding Information: Funding: This research was funded the BMBF cluster Rauchenzym, grant number 031B0586. The APC was funded by the Open Access fund of the Gottfried Wilhelm Leibniz Universität Hannover.

PY - 2021/5/14

Y1 - 2021/5/14

N2 - Traditional smoking generates not only the impact flavor compound 4-vinylguaiacol, but concurrently many unwanted and potent toxic compounds such as polycyclic aromatic hydrocarbons. Enzyme technology provides a solution without any side-product formation. A feruloyl esterase from Rhizoctonia solani (RspCAE) liberated ferulic acid from low-priced sugar beet fiber. Decarboxylation of ferulic acid to 4-vinylguaiacol was achieved by a second enzyme from Schizophyllum commune (ScoFAD). Both enzymes were covalently immobilized on agarose to enable reusability in a fixed-bed approach. The two enzyme cascades showed high conversion rates with yields of 0.8 and 0.95, respectively, and retained activity for nearly 80 h of continuous operation. The overall productivity of the model process with bed volumes of 300 µL and a substrate flow rate of 0.25 mL min −1 was 3.98 mg 4-vinylguaiacol per hour. A cold online solid phase extraction using XAD4 was integrated into the bioprocess and provided high recovery rates during multiple elution steps. Attempting to facilitate the bioprocess, a fused gene coding for the two enzymes and a set of different linker lengths and properties was constructed and introduced into Komagataella phaffii. Longer and rigid linkers resulted in higher activity of the fusion protein with a maximum of 67 U L −1.

AB - Traditional smoking generates not only the impact flavor compound 4-vinylguaiacol, but concurrently many unwanted and potent toxic compounds such as polycyclic aromatic hydrocarbons. Enzyme technology provides a solution without any side-product formation. A feruloyl esterase from Rhizoctonia solani (RspCAE) liberated ferulic acid from low-priced sugar beet fiber. Decarboxylation of ferulic acid to 4-vinylguaiacol was achieved by a second enzyme from Schizophyllum commune (ScoFAD). Both enzymes were covalently immobilized on agarose to enable reusability in a fixed-bed approach. The two enzyme cascades showed high conversion rates with yields of 0.8 and 0.95, respectively, and retained activity for nearly 80 h of continuous operation. The overall productivity of the model process with bed volumes of 300 µL and a substrate flow rate of 0.25 mL min −1 was 3.98 mg 4-vinylguaiacol per hour. A cold online solid phase extraction using XAD4 was integrated into the bioprocess and provided high recovery rates during multiple elution steps. Attempting to facilitate the bioprocess, a fused gene coding for the two enzymes and a set of different linker lengths and properties was constructed and introduced into Komagataella phaffii. Longer and rigid linkers resulted in higher activity of the fusion protein with a maximum of 67 U L −1.

KW - 4-vinylguaiacol

KW - Basidiomycetes

KW - Bioprocess

KW - Fusion protein

KW - Immobilization

KW - Smoke flavor

UR - http://www.scopus.com/inward/record.url?scp=85105728732&partnerID=8YFLogxK

U2 - 10.3390/catal11050631

DO - 10.3390/catal11050631

M3 - Article

VL - 11

JO - CATALYSTS

JF - CATALYSTS

SN - 2073-4344

IS - 5

M1 - 631

ER -