Novel Enzymatic Synthesis of 3-hydroxybutyric Acid Oligomerswith Inserted Lactobionic Acid Moieties

3-Hydroxybutyric acid and lactobionic acid yielded linear and cyclic oligomers in a lipase-catalyzedcondensation polymerization reaction, performed at 80°C in bulk and organic solvent systems. Novozyme435 was the most efficient biocatalyst, and a mixture of t-butanol and dimethylsulfoxide in 80:20 (v/v) ratioprovided the highest copolymer conversions. The highest degree of polymerization reached 7 in case ofcopolymers with inserted lactobionic acid moiety and 11 for the 3-hydroxybutyric acid homopolymer by-product

Lipase catalyzed synthesis of aromatic esters of sugar alcohols

Commercially available lipases (Candida antarctica lipase B, Novozyme 435, Thermomyces lanuginosus lipase, and Lipozyme TL IM), as well as sol-gel immobilized lipases, have been screened for their ability to acylate regioselectively xylitol, sorbitol, and mannitol with a phenolic ester in a binary mixture of t-butanol and dimethylsulfoxide. HPLC and MALDI-TOF MS analysis revealed the exclusive formation of monoesters for all studied sugar alcohols. The lipases immobilized by the sol-gel entrapment method proved to be efficient catalysts, leading to high conversions (up to 60%) in the investigated acylation reactions. From a sequence of silane precursors with different nonhydrolyzable groups in their structure, the presence of octyl and i-butyl group was most beneficial for the catalytic activity of sol-gel entrapped lipases in the studied process

Sol–gel immobilization of Alcalase from Bacillus licheniformis for application in the synthesis of C-terminal peptide amides

Alcalase 2.4L FG, a commercial preparation of Subtilisin A, was physically entrapped in glass sol–gel matrices using alkoxysilanes of different types mixed with tetramethoxysilane (TMOS). The materials were used for catalyzing C-terminal amidation of Z-Ala-Phe-OMe in a mixture of tert-butanol/DMF. From the screening of silane monomers in the sol–gel coating process, it was concluded that dimethyldimethoxysilane (DMDMOS) gave the best performance, and Alcalase immobilized therein exhibited the highest activity in the ammonolysis of Z-Ala-Phe-OMe. The percentage of protein immobilization was in the range of 68–98%, and total amidation activity of the immobilized Alcalase was up to 1.76 µmol/h/mg gel. We investigated the immobilization efficiency for a protein mass range of 2.8–9.7 mg per mmol total silanes, to determine the immobilization capacity of the biosilica support. The optimum enzyme loading capacity in the silica matrix was 115 mg/g dry silica xerogel (11.5%, w/w). The amount of the DMDMOS silicate was optimized by adjusting the molar ratio of silane mixture (DMDMOS and TMOS at 1:1). Biocatalyst sol–gel particles prepared at optimum immobilization conditions retained 100% of the original activity even after 14 cycles of repeated use. Reproducibility of the immobilization technique was also investigated by evaluating the catalytic efficiency of the obtained preparations. The thermal stability of the protease at 70 °C increased threefold upon entrapment in sol–gel materials, and twofold under storage for 50 days at ambient temperature