Next Article in Journal
The Antidiabetic Drug Metformin Inhibits the Proliferation of Bladder Cancer Cells in Vitro and in Vivo
Previous Article in Journal
A Simple Strategy for Development of Single Nucleotide Polymorphisms from Non-Model Species and Its Application in Panax
Int. J. Mol. Sci. 2013, 14(12), 24592-24602; doi:10.3390/ijms141224592
Essay

Characterization of a κ-Carrageenase from Marine Cellulophaga lytica strain N5-2 and Analysis of Its Degradation Products

1
, 1
, 1
, 2
 and 1,*
1 School of Life Science and Technology, Dalian University, Dalian 116622, China 2 School of Life Science, Dalian Nationalities University, Dalian 116600, China
* Author to whom correspondence should be addressed.
Received: 16 September 2013 / Revised: 4 December 2013 / Accepted: 6 December 2013 / Published: 17 December 2013
(This article belongs to the Section Bioinorganic Chemistry)
View Full-Text   |   Download PDF [832 KB, uploaded 19 June 2014]   |   Browse Figures

Abstract

A carrageenan-degrading marine Cellulophaga lytica strain N5-2 was isolated from the sediment of carrageenan production base. A κ-carrageenase (EC 3.2.1.83) with high activity was purified to electrophoretic homogeneity from the culture supernatant by a procedure of ammonium sulfate precipitation, dialyzing and gel filtration on SephadexG-200 and SephadexG-75. The purified enzyme was verified as a single protein on SDS-PAGE, and whose molecular weight was 40.8 kDa. The κ-carrageenase yielded a high activity of 1170 U/mg protein. For κ-carrageenase activity, the optimum temperature and pH were 35 °C and pH 7.0, respectively. The enzyme was stable at 40 °C for at least 2.5 h. The enzyme against κ-carrageenan gave a Km value of 1.647 mg/mL and a Vmax value of 8.7 μmol/min/mg when the reaction was carried out at 35 °C and pH 7.0. The degradation products of the k-carrageenase were analyzed by thin layer chromatography (TLC), high performance liquid chromatography (HPLC), electrospray ionization time-of-flight mass spectroscopy (ESI-TOF-MS) and 13C-NMR spectroscopy, and the results indicated that the enzyme was specific of the β-1,4 linkage and hydrolyzed κ-carrageenan into κ-neocarraoctaose-sulfate and κ-neocarrahexaose-sulfate first, and then broke κ-neocarraoctaose-sulfate into κ-neocarrabiose-sulfate and κ-neocarrahexaose-sulfate.
Keywords: κ-carrageenase; Cellulophaga lytica strain N5-2; characterization; degradation product κ-carrageenase; Cellulophaga lytica strain N5-2; characterization; degradation product
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Share & Cite This Article

Export to BibTeX |
EndNote


MDPI and ACS Style

Yao, Z.; Wang, F.; Gao, Z.; Jin, L.; Wu, H. Characterization of a κ-Carrageenase from Marine Cellulophaga lytica strain N5-2 and Analysis of Its Degradation Products. Int. J. Mol. Sci. 2013, 14, 24592-24602.

View more citation formats

Related Articles

Article Metrics

Comments

Citing Articles

[Return to top]
Int. J. Mol. Sci. EISSN 1422-0067 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert