Archaeon Aeropyrum pernix K1 produces biotechnologically interesting enzymes, and one amongs them is pernisine. Pernisine is a thermostable protease. It has potential applications in the chemical industry preferably as laundry additive or for degradation of the infective PrPSc which are harmful to health. We have successfully isolated and described an extracellular Ca2+-modulated serine protease named pernisine. We have obtained about half a milligram of pernisine from 6,4 liters of growth medium in which archaea were cultivated for two-days. Heterologous expression of pernisine was tested using rarer expression systems such as Streptomyces rimosus, Streptomyces lividans and well-established system like E. coli. Imunodetection technique confirmed recombinant pernisine when codon-optimized (pernisineCOS) sequence was used. In the case of codon-non-optimized sequence, pernisine expression was not detected.The enzymatic activity was confirmed by zymogramphy. With S. lividans we could not detect the expression of pernisine. Original (pernisine), codon-optimized (pernisineCOE) and mutant sequence (pernisineCOS355AE) of pernisine were used to assemble the constructs for E. coli. Expression of recombinant pernisine was detected only with those constructs which involved codon optimization of the pernisine, even though we used strains designed to troubleshoot the expression of rare codons. Identification of pernisine was confirmed by means of MS/MS and N-terminal sequencing. The purified enzyme was used for further physical and biochemical characterization.
D.09 Tutoring for postgraduate students
COBISS.SI-ID: 267092992In the patent application we described the method for degradation of protein aggregates and prion proteins in the different field of applications.
F.32 International patent
COBISS.SI-ID: 4197240Laboratory manual for the subject Physico-chemical Methods in Food Science in the Master Study Programme of Food Science. It comprehends following chapters: Densitometry, Water activity, Potentiometry. Conductometry, Spectrometry and Chromatography.
D.10 Educational activities
COBISS.SI-ID: 253834496Member of the research program took part and helped to create the documentary » Science doesn’t bite: Vitamins in the spring «. Some properties of vitamin C related to its content in foods and influence of food processing on its stability were presented through discussion and experiments. These topics are closely related to the research that is a part of the program.
D.10 Educational activities
COBISS.SI-ID: 3220337This book is intended for the students of Interdisciplinary doctoral study programme Biosciences at University of Ljubljana at elective subject »Antioxidants and methodology of antioxidant efficiency determination« (professor Helena Abramovič). This book deals with the problems of lipid oxidation in food, offers the knowledge associated with the use of antioxidants and informs with the methodology of the determination of antioxidant efficiency.
D.10 Educational activities
COBISS.SI-ID: 256199424