Purification and Characterization of a Fucoidanase (FNase S) from a Marine Bacterium Sphingomonas paucimobilis PF-1

The Search for enzyme activities that efficiently degrade marine polysaccharides is becoming an increasingly important area for both structural analysis and production of lower-molecular weight oligosaccharides. In this study, an endo-acting fucoidanase that degrades Miyeokgui fucoidan (MF), a sulfa...

Full description

Bibliographic Details
Main Authors: Woo Jung Kim, Joo Woong Park, Jae Kweon Park, Doo Jin Choi, Yong Il Park
Format: Article
Language:English
Published: MDPI AG 2015-07-01
Series:Marine Drugs
Subjects:
Online Access:http://www.mdpi.com/1660-3397/13/7/4398
id doaj-c526d9835845458cac19e14add3e059d
record_format Article
spelling doaj-c526d9835845458cac19e14add3e059d2020-11-24T22:38:01ZengMDPI AGMarine Drugs1660-33972015-07-011374398441710.3390/md13074398md13074398Purification and Characterization of a Fucoidanase (FNase S) from a Marine Bacterium Sphingomonas paucimobilis PF-1Woo Jung Kim0Joo Woong Park1Jae Kweon Park2Doo Jin Choi3Yong Il Park4Department of Biotechnology and The Biomaterial Engineering Research Center, The Catholic University of Korea, Bucheon, Gyeonggi-do 420-743, KoreaDepartment of Biotechnology and The Biomaterial Engineering Research Center, The Catholic University of Korea, Bucheon, Gyeonggi-do 420-743, KoreaDepartment of Pharmaceutical Science, University of Gachon, Yeonsu-dong, Yeonsu-gu, InCheon 406-799, KoreaDepartment of Biotechnology and The Biomaterial Engineering Research Center, The Catholic University of Korea, Bucheon, Gyeonggi-do 420-743, KoreaDepartment of Biotechnology and The Biomaterial Engineering Research Center, The Catholic University of Korea, Bucheon, Gyeonggi-do 420-743, KoreaThe Search for enzyme activities that efficiently degrade marine polysaccharides is becoming an increasingly important area for both structural analysis and production of lower-molecular weight oligosaccharides. In this study, an endo-acting fucoidanase that degrades Miyeokgui fucoidan (MF), a sulfated galactofucan isolated from the sporophyll (called Miyeokgui in Korean) of Undaria pinnatifida, into smaller-sized galactofuco-oligosaccharides (1000–4000 Da) was purified from a marine bacterium, Sphingomonas paucimobilis PF-1, by ammonium sulfate precipitation, diethylaminoethyl (DEAE)-Sepharose column chromatography, and chromatofocusing. The specific activity of this enzyme was approximately 112-fold higher than that of the crude enzyme, and its molecular weight was approximately 130 kDa (FNase S), as determined by native gel electrophoresis and 130 (S1), 70 (S2) and 60 (S3) kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimum pH and temperature of FNase S were pH 6.0–7.0 and 40–45 °C, respectively. FNase S activity was enhanced by Mn2+ and Na+ (115.7% and 131.2%), but it was inhibited by Ca2+, K+, Ba2+, Cu2+ (96%, 83.7%, 84.3%, and 89.3%, respectively), each at 1 mM. The Km, Vmax and Kcat values of FNase S on MF were 1.7 mM, 0.62 mg·min−1, and 0.38·S−1, respectively. This enzyme could be a valuable tool for the structural analysis of fucoidans and production of bioactive fuco-oligosaccharides.http://www.mdpi.com/1660-3397/13/7/4398fucoidanSphingomonas sp.fucoidanasegalactofuco-oligosaccharides
collection DOAJ
language English
format Article
sources DOAJ
author Woo Jung Kim
Joo Woong Park
Jae Kweon Park
Doo Jin Choi
Yong Il Park
spellingShingle Woo Jung Kim
Joo Woong Park
Jae Kweon Park
Doo Jin Choi
Yong Il Park
Purification and Characterization of a Fucoidanase (FNase S) from a Marine Bacterium Sphingomonas paucimobilis PF-1
Marine Drugs
fucoidan
Sphingomonas sp.
fucoidanase
galactofuco-oligosaccharides
author_facet Woo Jung Kim
Joo Woong Park
Jae Kweon Park
Doo Jin Choi
Yong Il Park
author_sort Woo Jung Kim
title Purification and Characterization of a Fucoidanase (FNase S) from a Marine Bacterium Sphingomonas paucimobilis PF-1
title_short Purification and Characterization of a Fucoidanase (FNase S) from a Marine Bacterium Sphingomonas paucimobilis PF-1
title_full Purification and Characterization of a Fucoidanase (FNase S) from a Marine Bacterium Sphingomonas paucimobilis PF-1
title_fullStr Purification and Characterization of a Fucoidanase (FNase S) from a Marine Bacterium Sphingomonas paucimobilis PF-1
title_full_unstemmed Purification and Characterization of a Fucoidanase (FNase S) from a Marine Bacterium Sphingomonas paucimobilis PF-1
title_sort purification and characterization of a fucoidanase (fnase s) from a marine bacterium sphingomonas paucimobilis pf-1
publisher MDPI AG
series Marine Drugs
issn 1660-3397
publishDate 2015-07-01
description The Search for enzyme activities that efficiently degrade marine polysaccharides is becoming an increasingly important area for both structural analysis and production of lower-molecular weight oligosaccharides. In this study, an endo-acting fucoidanase that degrades Miyeokgui fucoidan (MF), a sulfated galactofucan isolated from the sporophyll (called Miyeokgui in Korean) of Undaria pinnatifida, into smaller-sized galactofuco-oligosaccharides (1000–4000 Da) was purified from a marine bacterium, Sphingomonas paucimobilis PF-1, by ammonium sulfate precipitation, diethylaminoethyl (DEAE)-Sepharose column chromatography, and chromatofocusing. The specific activity of this enzyme was approximately 112-fold higher than that of the crude enzyme, and its molecular weight was approximately 130 kDa (FNase S), as determined by native gel electrophoresis and 130 (S1), 70 (S2) and 60 (S3) kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimum pH and temperature of FNase S were pH 6.0–7.0 and 40–45 °C, respectively. FNase S activity was enhanced by Mn2+ and Na+ (115.7% and 131.2%), but it was inhibited by Ca2+, K+, Ba2+, Cu2+ (96%, 83.7%, 84.3%, and 89.3%, respectively), each at 1 mM. The Km, Vmax and Kcat values of FNase S on MF were 1.7 mM, 0.62 mg·min−1, and 0.38·S−1, respectively. This enzyme could be a valuable tool for the structural analysis of fucoidans and production of bioactive fuco-oligosaccharides.
topic fucoidan
Sphingomonas sp.
fucoidanase
galactofuco-oligosaccharides
url http://www.mdpi.com/1660-3397/13/7/4398
work_keys_str_mv AT woojungkim purificationandcharacterizationofafucoidanasefnasesfromamarinebacteriumsphingomonaspaucimobilispf1
AT joowoongpark purificationandcharacterizationofafucoidanasefnasesfromamarinebacteriumsphingomonaspaucimobilispf1
AT jaekweonpark purificationandcharacterizationofafucoidanasefnasesfromamarinebacteriumsphingomonaspaucimobilispf1
AT doojinchoi purificationandcharacterizationofafucoidanasefnasesfromamarinebacteriumsphingomonaspaucimobilispf1
AT yongilpark purificationandcharacterizationofafucoidanasefnasesfromamarinebacteriumsphingomonaspaucimobilispf1
_version_ 1725715103438864384