Identification and determination of the intra- and extracellular aminopeptidase activity by synthetic L-Ala-, L-Tyr-, and L-Phe-beta-napthylamide.

A simple, rapid and straightforward procedure for identification and determination of intracellular and extracellular activity of aminopeptidases employing synthetic substrates beta-naphtylamides of L-Ala, L-Phe, and L-Tyr was used. Poppy cells (Papaver somniferum L.) permeabilized by Tween 80 were immobilized via crosslinking by glutaraldehyde. Glutaraldehyde immobilized poppy cells lost their viability and demonstrated significantly lower aminopeptidase activities than untreated control cells probably due to a damage to the enzyme active centre. Poppy cells immobilized by pectate and alginate have retained high activity of studied aminopeptidases. The culture medium (without cells) used for the identification and determination of extracellular enzyme activities retained 20-21%, whereas intracellular activities were estimated to be 79-80% of total enzyme activity. Thus the intracellular specific activity was 1.00-1.07 higher.

[Effect of a fungal elicitor on levels of sanguinarine and polyphenoloxidase activity in a suspension culture of Papaver somniferum L]. / Zmeny obsahu sanguinarínu a aktivity polyfenoloxidázy vplyvom fungálneho elicitora v suspenzných kultúrach maku siateho Papaver somniferum L.

The opium poppy (Papaver somniferum L.) is still a source for isolation of codeine and morphine. Cell cultures from this plant lose their ability to produce morphinans. Their major alkaloid is sanguinarine. The elicitation of the opium poppy cell cultures by fungal preparation lead to a nine-fold increase in the content of sanguinarine. The specific activity of polyphenoloxidase (PPO) was three-times higher in the elicited compared to the nonelicited cells. Two isoforms of PPO (Mr 63 kDa, 41 kDa) were identified in opium poppy cell cultures by PAGE. The number of PPO isoforms was not affected by elicitation. Phenyl-Sepharose CL-4B was used for affinity purification of PPO. In a single purification step the specific activity of PPO was enriched 14-fold.

Effects of substituted aryloxyaminopropanols on photosynthesis and photosynthesizing organisms.

The inhibitory activity of 22 substituted aryloxyaminopropanols having beta-lytic and local anaesthetic properties was studied from the view-point of their influences on photosynthesis in plant chloroplasts as well as growth and synthesis of chlorophyll in algae and wheat plants. The inhibitory activity increased significantly with the increasing length of alkyl-substituents of the aryloxyaminopropanol molecule. Less pronounced dependences were found with respect to the position of the substituent chain on benzene ring. The inhibitory activity was found to correlate well with the lipophilicity of the compounds studied.