Characterization of hydrocortisone bioconversion and 16S RNA gene in Synechococcus nidulans cultures.
A unicellular cyanobacterium, Synechococcus nidulans (Pringsheim) Komárek, was isolated from paddy-fields and applied in the biotransformation experiment of hydrocortisone (1). This strain has not been previously tested for steroid bioconversion. Fermentation was carried out in BG-11 medium supplemented with 0.05% substrate at 25 degrees C for 14 days of incubation. The obtained products were chromatographically purified followed by their characterization using spectroscopic methods. 11beta,17beta-dihydroxyandrost-4-en-3-one (2), 11beta-hydroxyandrost-4-en-3,17-dione (3), and androst-4-ene-3,17-dione (4) were the main bioproducts in the hydrocortisone bioconversion. The observed bioreaction characteristics were the side chain degradation of the substrate to prepare compounds (2) and (3) following the 11beta-dehydroxylation for accumulation of the compound (4). Time course study showed the accumulation of the product (2) from the second day of the fermentation and compounds (3) and (4) from the third day. All the metabolites reached their maximum concentration in seven days. Cyanobacterial 16S rRNA gene was also amplified by PCR. Sequences were amplified using the universal prokaryotic primers which amplify a approximately 400-bp region of the 16S rRNA gene. PCR products were sequenced to confirm their authenticity as 16S rRNA gene of cyanobacteria. The result of PCR blasted with other sequenced cyanobacteria in NCBI showed 99% identity to the 16S small subunit rRNA of seven Synechococcus species.
Rasoul-Amini
S
S
Department of Pharmaceutical Biotechnology and Pharmaceutical Sciences Research Center, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz 71345-1583, Iran.
Ghasemi
Y
Y
Morowvat
M H
MH
Ghoshoon
M B
MB
Raee
M J
MJ
Mosavi-Azam
S B
SB
Montazeri-Najafabady
N
N
Nouri
F
F
Parvizi
R
R
Negintaji
N
N
Khoubani
S
S
D001483:Base Sequence; D001711:Biotransformation; D006854:Hydrocortisone / Q000737:chemistry / Q000378:metabolism*; D007492:Iran; D008969:Molecular Sequence Data; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D012988:Soil Microbiology; D046940:Synechococcus / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; D006854:Hydrocortisone