Chimeric lactase capable of spontaneous and strong immobilization on cellulose and development of a continuous-flow system for lactose hydrolysis at high temperatures.

Abstract:

:Recombinant plasmids containing fusion proteins composed of two different modules were constructed and expressed in Escherichia coli. The modules encoded the lactase LacA (LacZ) from the thermophilic bacterium Thermoanaerobacter ethanolicus and the cellulase CelD, a cellulose-binding module (CBM) from Anaerocellum thermophilum. The CelD CBM provides a spontaneous and strong sorption of the fusion proteins onto a cellulose carrier. The enzymatic activities of both the free LacA protein and LacA-CelD CBM fusion proteins immobilized onto the cellulose carrier were assessed. The LacA activity of the fusion protein was dependent upon its position with respect to the CBM. The highest level of lactase activity and stability was observed when the lactase domain was localized at its N terminus. A continuous-flow column reactor of lactase immobilized on a cellulose carrier was constructed, and its activity was assessed. The lactose hydrolysis rate for a 150 mM (5%) solution at a flow rate of 1 reactor volume per min was 75%, which is a value optimal for further whey transformation into glucose/galactose syrup.

journal_name

Appl Environ Microbiol

authors

Velikodvorskaya GA,Tikhonova TV,Gurvits ID,Karyagina AS,Lavrova NV,Sergienko OV,Tashlitskii VN,Lunina NA,Lunin VG

doi

10.1128/AEM.01517-10

subject

Has Abstract

pub_date

2010-12-01 00:00:00

pages

8071-5

issue

24

eissn

0099-2240

issn

1098-5336

pii

AEM.01517-10

journal_volume

76

pub_type

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