A p-coumaroyl esterase from Rhizoctonia solani with a pronounced chlorogenic acid esterase activity.
Extracellular esterase activity was detected in submerged cultures of Rhizoctonia solani grown in the presence of sugar beet pectin or Tween 80. Putative type B feruloyl esterase (FAE) coding sequences found in the genome data of the basidiomycete were heterologously expressed in Pichia pastoris. Recombinant enzyme production on the 5-L bioreactor scale (Rs pCAE: 3245UL-1) exceeded the productivity of the wild type strain by a factor of 800. Based on substrate specificity profiling, the purified recombinant Rs pCAE was classified as a p-coumaroyl esterase (pCAE) with a pronounced chlorogenic acid esterase side activity. The Rs pCAE was also active on methyl cinnamate, caffeate and ferulate and on feruloylated saccharides. The unprecedented substrate profile of Rs pCAE together with the lack of sequence similarity to known FAEs or pCAEs suggested that the Rs pCAE represents a new type of enzyme. Hydroxycinnamic acids were released from agro-industrial side-streams, such as destarched wheat bran (DSWB), sugar beet pectin (SBP) and coffee pulp (CP). Overnight incubation of coffee pulp with the Rs pCAE resulted in the efficient release of p-coumaric (100%), caffeic (100%) and ferulic acid (85%) indicating possible applications for the valorization of food processing wastes and for the enhanced degradation of lignified biomass.
Institut für Lebensmittelchemie, Leibniz Universität Hannover, Callinstraße 5, 30167 Hannover, Germany. Electronic address: annabel.nieter@lci.uni-hannover.de.
Annabel
A
Nieter
Institut für Lebensmittelchemie, Leibniz Universität Hannover, Callinstraße 5, 30167 Hannover, Germany.
Sebastian
S
Kelle
Institut für Lebensmittelchemie, Leibniz Universität Hannover, Callinstraße 5, 30167 Hannover, Germany.
Diana
D
Linke
Institut für Lebensmittelchemie, Leibniz Universität Hannover, Callinstraße 5, 30167 Hannover, Germany.
Ralf G
RG
Berger
D002265:Carboxylic Ester Hydrolases / Q000235:genetics / Q000378:metabolism*; D003373:Coumaric Acids / Q000378:metabolism; D012232:Rhizoctonia / Q000201:enzymology* / Q000254:growth & development; D013379:Substrate Specificity
D003373:Coumaric Acids; D002265:Carboxylic Ester Hydrolases; C025374:chlorogenic acid esterase; C067271:trans-4-coumaroyl esterase
Agro-industrial by-products; Basidiomycota; Chlorogenic acid; Feruloyl esterase; Feruloylated saccharides; Heterologous expression; Pichia pastoris