产褐藻胶裂解酶海洋细菌的筛选鉴定及产酶条件优化  

Screening and identification of marine alginate lyase-producing bacteria and optimization of enzyme production conditions

作　　者：胡梦迪 黎耀祖 张晓勇 莫梅清 高向阳[1] HU Mengdi;LI Yaozu;ZHANG Xiaoyong;MO Meiqing;GAO Xiangyang(Guangdong Provincial Key Laboratory ofNutraceuticals and Functional Foods,College of Food Science,South China Agricultural University,Guangzhou 510642,China;College of Marine Sciences,South China Agricultural University,Guangzhou 510642,China)

机构地区：[1]华南农业大学食品学院广东省功能食品活性物重点实验室,广东广州510642 [2]华南农业大学海洋学院,广东广州510642

出　　处：《中国酿造》2025年第4期132-138,共7页China Brewing

基　　金：广东省自然科学基金(2023A1515011859)。

摘　　要：褐藻胶裂解酶大多来源于具有独特生存环境的海洋细菌,该研究以海藻酸钠为唯一碳源,采用透明圈法初筛及3,5-二硝基水杨酸(DNS)法测定酶活复筛,从56株海洋细菌中筛选高产褐藻胶裂解酶的菌株,通过形态学观察、生理生化试验和16S rDNA基因序列分析对筛选菌株进行鉴定,并以褐藻胶裂解酶活力为响应值,通过Plackett-Burman试验结合Box-Behnken响应面法优化筛选菌株产褐藻胶裂解酶的发酵条件。结果表明,筛选获得一株高产褐藻胶裂解酶的菌株I14,经鉴定为贝莱斯芽孢杆菌(Bacillus velezensis),其产褐藻胶裂解酶的最佳发酵条件为:海藻酸钠添加量5 g/L、可溶性淀粉添加量5 g/L、酵母粉添加量4.30 g/L、蛋白胨添加量10 g/L、NaCl添加量30 g/L、CaCl2添加量0.30 g/L、初始pH 6.0、发酵温度35℃、发酵时间24 h、装液量30 mL/100 mL。在此优化条件下,褐藻胶裂解酶活力达81.29 U/mL,是优化前的4.76倍。该研究为褐藻胶裂解酶的开发利用提供新的菌种选择,同时拓展该菌株在降解褐藻胶、生产多糖裂解酶等食品、生物领域的应用。Alginate lyases are mostly derived from marine bacteria with unique living environments.Using sodium alginate as the sole carbon source,a strain with high-yield alginate lyase from 56 strains of marine bacteria was screened using primary screening by transparent circle method and re-screening for enzyme activity determination by 3,5-dinitrosalicylic acid(DNS)method.The screened strain was identified by morphological observa-tion,physiological and biochemical tests and 16S rDNA gene sequence analysis,and using alginate lyase activity as the response value,the fermenta-tion conditions for the screened strain for alginate lyase production was optimized by Plackett-Burman experiment combined with Box-Behnken re-sponse surface method.The results showed that the strain I14 with high-yield alginate lyase was obtained,and it was identified as Bacillus velezensis,and the optimum fermentation conditions for alginate lyase production by the strain I14 were determined as follows:sodium alginate addition 5 g/L,soluble starch addition 5 g/L,yeast powder addition 4.30 g/L,peptone addition 10 g/L,NaCl addition 30 g/L,CaCl2 addition 0.30 g/L,initial pH 6.0,fermentation temperature 35 C,time 24 h,and liquid loading volume 30 ml/100 ml.Under these optimal conditions,the activity of alginate lyase reached 81.29 U/ml,which was 4.76 times that of before optimization.This study provided a new strain option for the development and utilization of alginate lyases,and expanded the application of strain I14 in food and biological fields such as degradation of alginate and production of polysaccha-ride lyases.

关 键 词：海洋细菌 筛选 鉴定 褐藻胶裂解酶 发酵条件 响应面法 

分 类 号：TS201.3[轻工技术与工程—食品科学]