Characterization of an extracellular ß-glucosidase from Dekkera bruxellensis for resveratrol production.

Polygonum cuspidatum is a widely grown crop with a rich source of polydatin (also called piceid) for resveratrol production. Resveratrol is produced from piceid via enzymatic cleavage of the sugar moiety of piceid. In this study, Dekkera bruxellensis mutants were selected based on their high p-nitrophenyl-ß-d-glucopyranoside and piceid conversion activities. The enzyme responsible for piceid conversion was a heterodimeric protein complex that was predominantly secreted to the extracellular medium and consisted of two subunits at an equal ratio with molecular masses of 30.5 kDa and 48.3 kDa. The two subunits were identified as SCW4p and glucan-ß-glucosidase precursor in D. bruxellensis. Both proteins were individually expressed in Saccharomyces cerevisiae exg1Δ mutants, which lack extracellular ß-glucosidase activity, to confirm each protein's enzymatic activities. Only the glucan-ß-glucosidase precursor was shown to be a secretory protein with piceid deglycosylation activity. Our pilot experiments of piceid bioconversion demonstrate the possible industrial applications for this glucan-ß-glucosidase precursor in the future.

The cysteine 703 to isoleucine or histidine mutation of the oxidosqualene-lanosterol cyclase from Saccharomyces cerevisiae generates an iridal-type triterpenoid.

The Cys703 to Ile or His mutation within Saccharomyces cerevisiae oxidosqualene-lanosterol cyclase ERG7 (ERG7(C703I/H)) generates an unusual truncated bicyclic rearranged intermediate, (8R,9R,10R)-polypoda-5,13E,17E,21-tetraen-3ß-ol, related to iridal-skeleton triterpenoid. Numerous oxidosqualene-cyclized truncated intermediates, including tricyclic, unrearranged tetracyclic with 17α/ß exocyclic hydrocarbon side chain, rearranged tetracyclic, and chair-chair-chair tricyclic intermediates (compounds 3-9), were also isolated from the ERG7(C703X) site-saturated mutations or the ERG7(F699T/C703I) double mutation, indicating the functional role of the Cys703 residue in stabilizing the bicyclic C-8 cation and the rearranged intermediate or interacting with Phe699, and opened a new avenue of engineering ERG7 for producing biological active agents.

Calorimetric studies and lessons on fires and explosions of a chemical plant producing CHP and DCPO.

Cumene hydroperoxide (CHP) has been used in producing phenol, dicumyl peroxide (DCPO) and as an initiator for synthesizing acrylonitrile-butadiene-styrene (ABS) resin by copolymerization in Taiwan. Four incidents of fire and explosion induced by thermal runaway reactions were occurred in a same plant producing CHP, DCPO and bis-(tert-butylperoxy isopropyl) benzene peroxide (BIBP). The fourth fire and explosion occurred in the CHP reactor that resulted in a catastrophic damage in reaction region and even spread throughout storage area. Descriptions on the occurrences of these incidents were assessed by the features of processes, reaction schemes and unexpected side reactions. Calorimetric data on thermokinetics and pressure were used for explaining the practical consequences or which the worst cases encountered in this kind of plant. Acceptable risk associated with emergency relief system design is vital for a plant producing organic peroxide. These basic data for designing an inherently safer plant can be conducted from adiabatic calorimetry. An encouraging deduction has been drawn here, these incidents may be avoided by the implementation of API RP 520, API RP 521, DIERS technology, OSHA 1910.119 and AIChE's CCPS recommended PSM elements.