Ma Shuang, Xiao Hou-Rong, Wu Qian-Qian, Pan Ren-Rui, Cai Jing-Min
College of Life Science, Anhui Agricultural University, Hefei 230036, China.
Guang Pu Xue Yu Guang Pu Fen Xi. 2008 Mar;28(3):590-3.
Fucoidanase was isolated and purified from marine fungus LD8 by solid state fermentation, extraction with citric acid buffer, acetone precipitatation and column chromatography on Sephadex G-100. A single band on PAGE shows that pretty pure fucoidanase has been obtained. FT-IR spectra and its derivation, self-deconvolution and curve-fitting methods were used to analyze the secondary structure of the fucoidanase. Composite bands of the amide I and amide III were studied by using Fourier self-deconvolution (FSD) with an enhancement factor of K = 2.2 and a half width of 20.2 cm(-1). The relative average fractions of alpha-helix, beta-sheet, random coil, beta-turn are 11.5%, 58.6%, 14.5% and 15.9%, respectively, according to amide I region, while the content of alpha-helix is 12%, beta-sheet 57.3%, random coil 14.5%, and beta-turn 16.3% on amide III region. In other words, both the conclusions were exactly consistent. All the above results show that beta-sheet was the dominant component, which is about 58%, and that beta-turn is about 15%, random coil 15%, and alpha-helix 12% at room temperature.
采用固态发酵、柠檬酸缓冲液提取、丙酮沉淀和Sephadex G - 100柱层析从海洋真菌LD8中分离纯化出岩藻糖酶。聚丙烯酰胺凝胶电泳(PAGE)上的单一条带表明已获得纯度较高的岩藻糖酶。利用傅里叶变换红外光谱(FT - IR)及其导数、自去卷积和曲线拟合方法分析岩藻糖酶的二级结构。采用增强因子K = 2.2、半高宽为20.2 cm⁻¹的傅里叶自去卷积(FSD)研究酰胺I和酰胺III的复合带。根据酰胺I区域,α - 螺旋、β - 折叠、无规卷曲、β - 转角的相对平均比例分别为11.5%、58.6%、14.5%和15.9%,而在酰胺III区域,α - 螺旋含量为12%,β - 折叠为57.3%,无规卷曲为14.5%,β - 转角为16.3%。也就是说,两个结论完全一致。上述所有结果表明,在室温下,β - 折叠是主要成分,约占58%,β - 转角约占15%,无规卷曲占15%,α - 螺旋占12%。