XXII. BIOCHEMICKÝ ZJAZD - Jesseniova lekárska fakulta

Posters
12.
HETEROLOGOUS EXPRESSION OF ROYAL JELLY APALBUMINS IN E. COLI
Tatiana Kraková, Jozef Šimúth and Katarína Bíliková
Department of Molecular Apidology, Institute of Molecular Biology, SAS Bratislava
The honeybee royal jelly (RJ) contains many valuable components and biologically active
proteins and peptides. Major proteins of RJ, designated as apalbumins belong to a protein
family consisting of nine members with M r
of 49-87 kDa. Apalbumin1, apalbumin2 and
apalbumin3 account for 90% of the RJ protein content and are 88% identical in amino
acid sequence. Minor RJ proteins are mainly homologues of apalbumins, antimicrobial
peptides and enzymes. RJ proteins display multifunctional features. For example, apalbumin1,
apalbumin2 stimulated tumor necrosis factor alpha release. We have focused our
attention to the molecular characterization of apalbumin2, the second most abundant
protein of RJ. Apalbumin2 is a basic protein of 52.7 kDa with eight isoelectric variants in
the range of pI 7.5-8.5. For first time we purified royal jelly and characterized minority
homologue of apalbumin2 named as apalbumin2a (48.6 kDa) with antibiotics activity
against P. larvae, the inducer of American foulbrood of honeybee colony.
The aim of our study was a biotechnological preparation of apalbumin1 and apalbumin2
and their homologues by heterologous expression in E. coli with the goal of study their
antibiotic and immunostimulatory properties. For these purposes we cloned appropriate
cDNA or their of PCR fragments into pQE30 and pET28b(+) expression vectors and
transformed into E. coli DH5α and/or BL21-CodonPlus(DE3)-RIL. We present the molecular
properties of recombined apalbumins and their homologues as well.
XXII. Biochemistry Congress, Martin
127