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

التفاصيل البيبلوغرافية
العنوان: Characterization of a κ-Carrageenase from Marine Cellulophaga lytica strain N5-2 and Analysis of Its Degradation Products
المؤلفون: Ziang Yao, Feifei Wang, Zheng Gao, Liming Jin, Haige Wu
المصدر: International Journal of Molecular Sciences, Vol 14, Iss 12, Pp 24592-24602 (2013)
بيانات النشر: MDPI AG, 2013.
سنة النشر: 2013
المجموعة: LCC:Biology (General)
LCC:Chemistry
مصطلحات موضوعية: κ-carrageenase, Cellulophaga lytica strain N5-2, characterization, degradation product, Biology (General), QH301-705.5, Chemistry, QD1-999
الوصف: 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.
نوع الوثيقة: article
وصف الملف: electronic resource
اللغة: English
تدمد: 1422-0067
Relation: http://www.mdpi.com/1422-0067/14/12/24592; https://doaj.org/toc/1422-0067
DOI: 10.3390/ijms141224592
URL الوصول: https://doaj.org/article/6939208829064fe9a01d64c9775c8646
رقم الأكسشن: edsdoj.6939208829064fe9a01d64c9775c8646
قاعدة البيانات: Directory of Open Access Journals
الوصف
تدمد:14220067
DOI:10.3390/ijms141224592