厨余中产淀粉酶芽孢杆菌的筛选、鉴定及酶学性质研究  

Isolation and identification of amylase producing Bacillus from kitchen waste and its enzymatic properties

作　　者：张林吉[1] 张海涛[1] 李云龙[1] 迟兰[1] 李心海[1] 开昌春 任士飞[1] ZHANG Lin-ji;ZHANG Hai-tao;LI Yun-long;CHI Lan;LI Xin-hai;KAI Chang-chun;REN Shi-fei

机构地区：[1]徐州生物工程职业技术学院,江苏徐州221006 [2]连云港市欧亚饲料有限公司,江苏连云港222314

出　　处：《饲料研究》2021年第10期69-73,共5页Feed Research

基　　金：徐州市科技局农业与社会发展资助项目(项目编号:KC19200);江苏省高等职业院校专业带头人高端研修资助项目(项目编号:2020GRFX096);2019年江苏省政策引导类计划(苏北科技专项)(项目编号:XZ-SZ201924);2020年江苏高校“青蓝工程”资助项目;江苏省高等学校自然科学研究项目(项目编号:18KJD240001)。

摘　　要：试验筛选厨余中具有高效降解淀粉功能的芽孢杆菌。采用可溶性淀粉培养基进行菌株的富集及分离培养,获得13株产淀粉酶的芽孢杆菌,测定其透明圈直径与菌落直径比值2.9^6.3,其中S8菌株和S10菌株产酶能力较高,总酶活达到283.6 U/mL和329.7 U/mL。通过形态、生化特征及16S rRNA基因序列分析,鉴定S8菌株为枯草芽孢杆菌,S10菌株为解淀粉酶芽孢杆菌。S10菌株酶学性质显示,该淀粉酶以β-淀粉酶为主,占比为69.1%,总淀粉酶最适反应温度和pH值为35℃和5.6。该酶在35^40℃之间具有很好的稳定性,高温耐受性差。适宜pH值下,4 h残余酶活依然在70%以上。Ca^(2+)和Mn^(2+)对酶活有较强的激活作用,Zn^(2+)、Cu^(2+)和Fe^(3+)对酶活有明显的抑制作用。The experiment obtain Bacillus with high starch degradation function in kitchen waste.13 strains of amylaseproducing Bacillus strains were obtained by enrichment and isolation culture with soluble starch medium;the ration of hydrolysis zone diameter to colony diameter was 2.9~6.3.Strain S8 and strain S10 had high amylase-producing activity,which the total enzyme activity reached 283.6 U/mL and 329.7 U/mL.Through the morphological characteristics,physiological and biochemical characteristics and 16S rRNA gene phylogenetic analysis,strain S8 was identified as Bacillus subtilis and strain S10 was Bacillus amyloliquefaciens.The enzyme properties of strain S10 showed thatβ-amylase was the main component,accounting for 69.1%.The optimum temperature and pH value of the enzyme reaction were 35℃and 5.6.The enzyme have good thermal stability at 35~40℃and poor tolerance to high temperature.The residual enzyme activity was still above 70%in suitable pH buffer for 4 h.The enzyme activity was activated by metal ion of Ca^(2+),Mn^(2+)and restrained by Zn^(2+),Cu^(2+)and Fe^(3+).

关 键 词：厨余 枯草芽孢杆菌 解淀粉酶芽孢杆菌 淀粉酶 酶活 

分 类 号：S816.3[农业科学—饲料科学]