Screening of Commercial Enzymes for Transfructosylation of Tyrosol: Effect of Process Conditions and Reaction Network

Abstract

Enzymatic fructosylation of organic acceptors other than saccharides brings new possibilities to synthesize molecules that do not exist in nature. The introduction of fructosyl moiety may lead to glycosides possessing enhanced physicochemical and bioactive properties which could be useful in the pharmaceutical and cosmetic industry. In this work, the regioselective synthesis of tyrosol β‐d‐fructofuranoside (TF) catalyzed by β‐fructofuranosidase is investigated. In the first step, 32 commercial enzyme preparations are screened for fructoside‐hydrolyzing activity. The most active preparations are subsequently examined for fructofuranosyl transfer from sucrose to tyrosol. The best candidate, Novozym 188, is chosen to study the effect of reaction conditions on the product formation in a batch reactor. The effects of substrate concentration, temperature, pH, time, and enzyme dosage on the concentration of TF produced are studied using the design of experiments methodology. The maximal product concentration of 3.8 g L−1 is achieved for the sucrose concentration of 1.5 m, tyrosol concentration of 29 g L−1, temperature of 41 °C, and pH 5.1. Besides the main transfructosylation reaction between sucrose and tyrosol, several side reactions take place. A reaction network includes also the formation of fructooligosaccharides and the hydrolysis of sucrose and all reaction products.

Production of Fructooligosaccharides Using a Commercial Heterologously Expressed Aspergillus sp. Fructosyltransferase

“…The period of the fast increase was stopped at this conversion value. The course of sucrose conversion gives an impression of the existence of apparent pseudo-equilibrium at 92%, which is similar to those in our earlier papers [11,16]. However, this longer experiment showed that the sucrose conversion degree grew slowly to 94% during the period of the subsequent transformation of lower FOS oligomers into higher ones.…”

“…Its specific activity can be directly compared with those of the preparations examined in the paper by Hollá et al, since the same activity determination method was used [16]. Seqenzym ® FT has an about 30 times higher specific activity level than Pectinex Ultra SP-L does and an about 70 times higher level than that of Viscozyme L. The only commercial preparation that could have had the specific activity of the same order of magnitude as Seqenzym ® FT is Rohapect CM [15].…”

“…The latter ones facilitate the biocatalytic process by excluding the tedious cultivation and purification step to obtain FTases with sufficient transfructosylation activity [14,15]. Many commercial enzyme preparations (pectinases, polygalacturonases, and glucanases) also have the potential to produce FOS because of their FTase side activity [15][16][17].…”

One-step Removal of Proteases in a Commercial Lactase Preparation from <i>Kluyveromyces lactis</i> Using an Anion-exchange Membrane

“…Most industrial enzyme preparations include not only the main target enzyme but also other contaminating ones. When these preparations are used for food processing, proteases from microbes often lead to contamination (Fia et al, 2014, Hollá et al, 2019. Lactase (β-galactosidase, EC 3.2.1.23) is a glycoside hydrolase enzyme that catalyzes the hydrolysis of lactose to glucose and galactose.…”