Putative role of a Streptomyces coelicolor-derived α-mannosidase in deglycosylation and antibiotic production.
SCO0948 was found to be the single open reading frame annotated to encode an α-mannosidase (AM1) in Streptomyces coelicolor M145. To characterize the protein, we overexpressed SCO0948 in Escherichia coli BL21(DE3). Recombinant AM1, with a molecular weight of 110 kDa, exhibited α-mannosidase activity toward 4-nitrophenyl-α-D-mannopyranoside with a K m of 4.61 mM, a V(max) of 101.6 mM/min, and a specific activity of 47.96 U/mg. Treatment of ovalbumin, a glycoprotein, with AM1 resulted in partial deglycosylation, as assessed by glycostaining and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The S. coelicolor deletion mutant for SCO0948 failed to produce α-mannosidase activity, confirming AM1 as the only α-mannosidase in S. coelicolor M145. Interestingly, the deletion mutant and a complementation strain produced lower levels of the antibiotics actinorhodin and undecylprodigiosin in glucose minimal media. The results indicate that AM1 as an α-mannosidase influences deglycosylation and antibiotic production in S. coelicolor M145.
Department of Microbial Engineering, College of Engineering, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul, 143-701, South Korea.
Thangamani
T
Rajesh
Jong-Min
JM
Jeon
Eunjung
E
Song
Hae-Min
HM
Park
Hyung Min
HM
Seo
Hyun-Joong
HJ
Kim
Da-Hye
DH
Yi
Yong-Hyun
YH
Kim
Kwon-Young
KY
Choi
Yun-Gon
YG
Kim
Hyung-Yeon
HY
Park
Yoo Kyung
YK
Lee
Yung-Hun
YH
Yang
D000900:Anti-Bacterial Agents / Q000096:biosynthesis / Q000737:chemistry; D004926:Escherichia coli; D005960:Glucosides / Q000737:chemistry; D006031:Glycosylation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D048372:Streptomyces coelicolor / Q000201:enzymology*; D043323:alpha-Mannosidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D000900:Anti-Bacterial Agents; D005960:Glucosides; D011994:Recombinant Proteins; C019502:4-nitrophenyl alpha-glucoside; D043323:alpha-Mannosidase