Measurement of in situ halophilic glyceraldehyde-3-phosphate dehydrogenase activity from the permeabilized cells of archaebacterium Haloarcula vallismortis

The Journal of General and Applied Microbiology

Anita R. Patil¹  Stanislaus F. D'Souza¹ 

[1] Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre

关键词: cell permeabilization;    detergents;    glyceraldehyde-3-phosphate dehydrogenase;    Haloarcula vallismortis;    halophilic;    in situ activity;    lysozyme;    papain;   

DOI  :  10.2323/jgam.43.163

学科分类：微生物学和免疫学

来源: Applied Microbiology, Molecular and Cellulrar Biosciences Research Foundation

【 摘 要 】

The cells of Haloarcula vallismortis, an extreme halophilic archaebacterium, were permeabilized by various chemical, physical, and biological treatments. Biological permeabilization by lysozyme and papain showed effective results as observed by studying the in situ activity of halophilic glyceraldehydes -3-phosphate dehydrogenase (hGAPDH) as the model enzyme. Detergents N-cetyl-N, N, N-trimethyl ammonium bromide (CTAB) and digitonin also showed significant results. Other strains of halobacteria could also be permeabilized by lysozyme. The cell morphology did not show any significant change after permeabilization as observed by phase contrast microscopy. The enzyme characteristics of hGAPDH were studied in situ using permeabilized H. vallismortis cells. The properties, like optimum pH, Km for GAP and NAD+, inhibition by heavy metals, sulphydryl reagents, and other compounds, showed remarkable similarity with those studied in vitro.

【 授权许可】

Unknown