海洋来源低温果胶酶产生菌筛选、鉴定与酶学性质研究  

Screening,Identification and Enzymatic Properties Study of Coldadapted Marine Pectinase-Producing Bacteria

作　　者：赵子琪 刘烨瑀 于爽 迟雪梅[2,3] 金书寒 陈立功[3,4] 张庆芳 王晓辉 ZHAO Ziqi;LIU Yeyu;YU Shuang;CHI Xuemei;JIN Shuhan;CHEN Ligong;ZHANG Qingfang;WANG Xiaohui(School of Medicine,Dalian University,Dalian 116000,China;Liaoning Marine Microbiology Engineering Technology Research Center,Dalian 116622,China;College of Life Sciences and Technology,Dalian University,Dalian 116622,China;Shenzhen New Industry Biomedical Engineering Company Limited,Shenzhen 518000,China)

机构地区：[1]大连大学医学院,辽宁大连116000 [2]辽宁省海洋微生物工程技术研究中心,辽宁大连116622 [3]大连大学生命科学与技术学院,辽宁大连116622 [4]深圳市新产业生物医学工程股份有限公司,广东深圳518000

出　　处：《食品工业科技》2024年第10期133-140,共8页Science and Technology of Food Industry

基　　金：中央引导地方科技发展资金项目(110013);国家自然科学基金青年科学基金项目(31500039);大连市青年科技之星项目(2017RQ155)。

摘　　要：目的:从海洋底泥样品中筛选能低温降解果胶的新颖菌株,发掘具有潜在产业化价值的果胶酶。方法:以果胶为唯一碳源,利用透明圈法和3,5-二硝基水杨酸法(DNS法)从大连渤海湾海泥中筛出一株高产果胶酶菌株。对其进行形态学、生理生化特征分析以及16S rDNA序列测定,BLAST同源基因比对后构建系统发育树,并进一步研究其酶学特性。结果:筛选高产果胶酶菌株命名为LY-1,通过16S rDNA序列分析并结合生理生化鉴定确定该菌株为橄榄灰链霉菌(Streptomyces olivogriseus)。菌株LY-1在25℃、180 r/min的条件下培养至24 h时,果胶酶的酶活力为30.56 U/mL。最适酶pH为8.0,最适酶温度为20℃。此外,该酶在60℃孵育2 h仍保持70%以上的酶活性,在pH7.0~9.0范围内稳定性良好,酶活性稳定维持在90%以上。金属离子Cu^(2+)、Ba^(2+)、Mg^(2+)、Ca^(2+)、K+、Co^(2+)促进该酶活性,Mn^(2+)、Zn^(2+)、Hg^(2+)、Na^(+)则抑制该酶活性。结论:菌株LY-1所产的果胶酶具有应用于产业化商用果胶酶的潜力。Objective:To select novel strains that could degrade pectin at low temperature from marine sediment samples and discover pectinase with potential industrial value.Methods:Take pectin as the only carbon source,screening a highyielding pectinase strain from sea mud in Bohai Bay,Dalian with the transparent circle method and the 3,5-dinitrosalicylic acid method(DNS method).It analyzed the morphological,physiological and biochemical characteristics,and determined the 16S rDNA sequence.After comparing BLAST homologous gene,it built up a phylogenetic tree,and further studied the enzymatic properties.Results:It screened the strain with high pectinase yield which was named LY-1.Then it analyzed the 16S rDNA sequence and determined that this strain was Streptomyces olivogriseus from the physiological and biochemical index.When strain LY-1 was cultured at 25℃and 180 r/min for 24 hours,the enzyme activity of pectinase was 30.56 U/mL.It showed that the optimal enzyme pH was 8.0,and the optimal enzyme temperature was 20℃.Furthermore,the enzymatic activity could still be maintained more than 70%when it was incubated at 60℃for 2 hours,and remained stable in the pH range of 7.0~9.0 with enzyme activity at over 90%.The metal ions Cu^(2+),Ba^(2+),Mg^(2+),Ca^(2+),K+,and Co^(2+)could promote the activity of this enzyme,while Mn^(2+),Zn^(2+),Hg^(2+),and Na^(+)would inhibit its activity.Conclusion:The pectinase produced by strain LY-1 would have the potential to be applied to industrial commercial pectinase.

关 键 词：果胶酶 筛选 鉴定 酶学性质 

分 类 号：TS201.3[轻工技术与工程—食品科学]