Study of gherkin lactase in cell culture and in seedlings.
A synthetic substrate replacing lactose has facilitated application of a simple, rapid and sensitive method for the identification and determination of extracellular and intracellular gherkin lactase. The intracellular enzyme activity was estimated from the cell suspension, while the extracellular enzyme activity was established within the cell free cultivation medium. A suspension of gherkin cells was permeabilized by Tween 20, or Tween 80, or hexadecyltrimethyl ammonium bromide, or hexadecylpyridinium chloride or ethanol added one at a time and then immobilized by glutaraldehyde. The highest lactase activity was at pH 4.8 at a temperature of 55°C. The hydrolysis of substrate was linear for 4.5h and reached 60% conversion. The cells had high lactase activity and good stability. During long-term storage they demonstrated convenient physico-mechanical properties.
Stano
Ján
J
Garden of Medicinal Plants, Faculty of Pharmacy, Comenius University, Bratislava, Slovak Republic.
Siekel
Peter
P
Neubert
Klaus
K
Mičieta
Karol
K
D002470:Cell Survival; D002478:Cells, Cultured; D018553:Cucumis sativus / Q000166:cytology* / Q000201:enzymology*; D004789:Enzyme Activation; D004795:Enzyme Stability; D006868:Hydrolysis; D043322:Lactase / Q000302:isolation & purification / Q000378:metabolism*; D036226:Seedlings / Q000166:cytology* / Q000201:enzymology*; D013696:Temperature
D043322:Lactase