小龙虾肠道产果胶酶菌株的筛选、鉴定和酶学性质研究  

Study on screening,identification,and enzymatic properties of pectinase-producing strains in crayfish(Procambarus clarkii)intestine

作　　者：冯光志 田尹琼 江鑫 孙正 刘欣 杨晗月 张智 石玉 FENG Guangzhi;TIAN Yinqiong;JIANG Xin;SUN Zheng;LIU Xin;YANG Hanyue;ZHANG Zhi;SHI Yu

机构地区：[1]武汉设计工程学院食品与生物科技学院,湖北武汉430205

出　　处：《饲料研究》2025年第3期114-119,共6页Feed Research

基　　金：湖北省高等学校优秀中青年科技创新团队计划项目(项目编号:T2021047);湖北省教育厅科研计划项目(项目编号:B2020303、B2021375)。

摘　　要：试验旨在筛选小龙虾肠道产果胶酶的菌株,测定其酶学性质。采用高通量测序分析小龙虾肠道菌群结构,采用纯培养法从小龙虾肠道筛选产果胶酶菌株,对高活性菌株进行分子鉴定及酶学性质研究。结果显示,从小龙虾肠道中筛选到产果胶酶菌株7株,其中菌株C21产酶最高,形态学结合16S rDNA分子鉴定结果表明菌株C21为类芽孢杆菌(Paenibacillus terreus)。菌株C21所产果胶酶的最适pH值为11.0,最适反应温度为55℃;在pH值8~12条件下放置24 h后相对酶活保留80%以上,在温度为20~40℃范围内保温3 h,其果胶酶相对酶活仍保留80%以上;Mg^(2+)、Fe^(2+)、Al^(3+)、Cu^(2+)对果胶酶有促进作用,其中Mg^(2+)和Cu^(2+)可明显提高果胶酶的相对酶活,Ca^(2+)对酶无明显作用,Zn^(2+)和Mn^(2+)对果胶酶具有抑制作用,其中Mn^(2+)可使相对酶活降至12.87%。高通量测序结果表明,在小龙虾全肠中门水平中包括:变形菌门(Proteobacteria)、软壁菌门(Tenericutes)、厚壁菌门(Firmicutes)、拟杆菌门(Bacteroidetes)、放线菌门(Actinobacteria)等;在属水平中包括:伯克氏菌属(Burkholderia)、棍状厌氧菌属(Anaerorhabdus)、不动杆菌属(Acinetobacter)、假单胞菌属(Pseudomonas)、金黄杆菌属(Chryseobacterium)、芽孢杆菌属(Bacillus)等。研究表明,芽孢杆菌在小龙虾肠道中的数量适中,类芽孢杆菌C21具有较高的产果胶酶能力,可作为益生菌开发小龙虾肠道微生物源高效饲料。The experiment aimed to screen for pectinase-producing strains from the intestinal tract of crayfish and characterize their enzymatic properties.High-throughput sequencing was used to analyze the microbial community structure of crayfish intestine.Pure culture method was used to screen pectinase-producing bacteria from crayfish intestine,and molecular identification and enzymatic property researchs were carried out on high-activity strains.The results showed that seven strains of pectinase-producing bacteria were screened from crayfish intestine,among which strain C21 had the highest enzyme production.Morphological and 16S rDNA molecular identification results showed that strain C21 was Paenibacillus terreus.The optimal pH value of pectinase produced by strain C21 was 11.0,and the optimal reaction temperature was 55℃.The pectinase retained over 80%of its relative enzymatic activity after 24 hours under pH 8~12 conditions.When maintained at 20~40℃for 3 hours,it also preserved more than 80%relative activity.Metal ion analysis revealed that Mg^(2+),Fe^(2+),Al^(3+),and Cu^(2+)enhanced pectinase activity,with Mg^(2+)and Cu^(2+)demonstrating particularly significant activation effects.Ca^(2+)showed no observable impact on the enzyme,while Zn^(2+)and Mn^(2+)exhibited inhibitory effects,notably reducing the relative enzymatic activity to 12.87%in the case of Mn^(2+).The results of high-throughput sequencing showed that in the whole intestine of crayfish,at the phylum level,it included Proteobacteria,Tenericutes,Firmicutes,Bacteroidetes,Actinobacteria,etc.At the genus level,it included Burkholderia,Anaerorhabdus,Acinetobacter,Pseudomonas,Chryseobacterium,Bacillus,etc.The study indicates that the population of Bacillus in the intestines of crayfish is moderate,and Paenibacillus C21 had a high ability to produce pectinase,which could be developed as a probiotic for the development of efficient feed from crayfish intestinal microbe sources.

关 键 词：小龙虾肠道 果胶酶 筛选 酶学性质 菌群结构 

分 类 号：S816.7[农业科学—饲料科学]