产岩藻多糖酶菌株的筛选及其酶解制备低分子质量岩藻多糖的研究  

Screening of fucoidanase-producing strains and preparation of low molecular weight fucoidan by enzymatic hydrolysis

作　　者：王聪聪 周剑丽 顾秋亚[1] 杨文华 张文帅 余晓斌[1] WANG Congcong;ZHOU Jianli;GU Qiuya;YANG Wenhua;ZHANG Wenshuai;YU Xiaobin(Key Laboratory of Industrial Biotechnology,Ministry of Education,School of Biotechnology,Jiangnan University,Wuxi 214122,China;Province Key Laboratory of Fermentation Engineering and Biopharmacy,School of Liquor and Food Engineering,Guizhou University,Guiyang 550025,China)

机构地区：[1]江南大学生物工程学院,工业生物技术教育部重点实验室,江苏无锡214122 [2]贵州大学酿酒与食品工程学院,发酵工程与生物制药重点实验室,贵州贵阳550025

出　　处：《食品与发酵工业》2022年第23期49-56,共8页Food and Fermentation Industries

基　　金：国家轻工技术与工程一流学科自主课题资助项目(LITE2018-11);贵州省黔科合支撑项目([2020]1Y144)。

摘　　要：以酒曲为原料,通过唯一碳源法和酶活力检测筛选得到一株产岩藻多糖酶的菌株,其酶活力为0.414 U/mL,经形态学和分子生物学鉴定为谢瓦氏曲霉(Aspergillus chevalier)。利用该菌株来源的岩藻多糖酶水解制备分子质量<10 kDa的低分子质量岩藻多糖(low molecular weight fucoidan,LMWF),并通过单因素和正交实验对酶解条件进行优化。结果显示,在底物质量浓度15 g/L,pH 6.0,酶解温度55℃,酶解时间24 h的条件下,LMWF得率最高,为52.85%。对酶解得到的LMWF进行分级纯化,共获得3种不同分子质量的产物,其中F1分子质量为1250 Da,F2为2505 Da,F3为8227 Da。抗氧化结果显示,3种产物均具有优于岩藻多糖的抗氧化活性。表明利用A.chevalieri WA1207来源的岩藻多糖酶水解制备分子质量<10 kDa的LMWF具有可行性,研究结果为后续解析岩藻多糖结构、探究岩藻多糖功能与结构之间的关系奠定了基础。In this study,a new fucoidanase-producing strain was screened from Daqu by using fucoidan as the only carbon source with enzyme activity detection,which was identified as Aspergillus chevalier by morphology and molecular biology and the fucoidanase activity of this fungi was 0.414 U/mL.The low molecular weight fucoidan(<10 kDa)was prepared by hydrolysis with fucoidanase,and the enzymatic hydrolysis conditions were optimized by single factor and orthogonal experiments.The results showed that under the conditions of substrate concentration of 15 g/L,pH 6.0,the enzymatic hydrolysis temperature of 55°C,and the enzymatic solution time of 24 h,the highest yield of low molecular weight fucoidan was up to 52.85%.The low molecular weight fucoidan obtained by enzymatic hydrolysis was purified in a graded manner,and three products with different molecules were obtained:F1 was 1250 Da,F2 was 2505 Da,and F3 was 8227 Da.The antioxidant results showed that three products have a good antioxidant activity than fucoidan,this indicated that it is feasible to prepare low molecular weight fucoidan with molecular weight less than 10 kDa by hydrolysis of fucoidanase from A.chevalieri WA1207,which lays the foundation for the subsequent analysis of fucoidan structure and investigation of the relationship between function and structure.

关 键 词：岩藻多糖酶 低分子质量岩藻多糖 菌株筛选 抗氧化 真菌 

分 类 号：TQ920.1[轻工技术与工程—发酵工程]