Automation of enzymatic hydrolysis by use of continuous flow methods involving immobilized biocatalysts

✍ Scribed by J.M. Fernández-Romero; M.D. Luque de Castro; M. Valcárcel

• Publisher: Elsevier Science
• Year: 1992
• Tongue: English
• Weight: 581 KB
• Volume: 264
• Category: Article
• ISSN: 0003-2670
• DOI: 10.1016/0003-2670(92)87015-d

✦ Synopsis

Two flow-lryectlon methods for on-hne enzymatic hydrolysis usmg unmobdlzed bmcatalysts are proposed and apphed to the hydrolysis of 4-nitrophenyl-/3-o-glucuromde catalysed by fi-o-glucuronidase Momtormg 1s performed via the two hydrolysis products of glucurorudes the aglucone (specific spectrophotometnc method) and glucuromc acid (umversal chenulummescence method) The spectrophotometic method afforded a linear determination range of l-700 PM wrlth excellent preaslon, R S D < 15%, and a sampling frequency of 130 h-' The chemdununescence method provided a linear determmatlon range of 0 05-5 5 mM, an R S D between 148 and 3 07% for concentrations of 0 17 and 172 mM, respectively, and a sample throughput of 25 h-' The yield of the enzymatic process and the stabfity of the enzyme reactor were determmed by using urme samples Two fully automated approaches to on-hne hydrolysis and collection and momtonng of the hydrolysate are also proposed