Formaldehyde (FA)-containing indoor air has a negative effect on human health and should be removed by intensive ventilation or by catalytic conversion to non-toxic products. FA can be oxidized by alcohol oxidase (AOX) taking part in methanol metabolism of methylotrophic yeasts. In the present work, AOX isolated from a Hansenula polymorpha C-105 mutant (gcr1 catX) overproducing this enzyme in glucose medium, was tested for its ability to oxidize airborne FA. A continuous fluidized bed bioreactor (FBBR) was designed to enable an effective bioconversion of airborne FA by AOX or by permeabilized mutant H. polymorpha C-105 cells immobilized in calcium alginate beads. The immobilized AOX having a specific activity of 6-8Umg -1 protein was shown to preserve 85-90% of the initial activity. The catalytic parameters of the immobilized enzyme were practically the same as for the free enzyme (k cat/K m was 2.35×10 3M -1s -1 vs 2.89×10 3M -1s -1, respectively). The results showed that upon bubbling of air containing from 0.3 up to 18.5ppm FA through immobilized AOX in the range of 1.3-26.6Ug -1 of the gel resulted in essential decrease of FA concentration in the outlet gas phase (less than 0.02-0.03ppm, i.e. 10-fold less than the threshold limit value). It was also demonstrated that a FBBR with immobilized permeabilized C-105 cells provided more than 90% elimination of airborne FA. The process was monitored by a specially constructed enzymatic amperometric biosensor based on FA oxidation by NAD + and glutathione-dependent formaldehyde dehydrogenase from the recombinant H. polymorpha Tf 11-6 strain.
Bibliographical noteFunding Information:
This work was financially supported by the Ministry of Science and Education of Ukraine (Grant M/157-2009 ), by the Ministry of Science, Culture and Sport of the State of Israel (Grant 1236 ) and in part, by the Samaria and Jordan Rift Valley Regional R&D Center (Israel) and by the Research Authority of the Ariel University Center of Samaria (Israel).
- Alcohol oxidase
- Biosensor assay
- Enzyme immobilization
- Fluidized bed bioreactor
- Recombinant formaldehyde dehydrogenase