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对解淀粉芽孢杆菌(Bacillus amyloliquefaciens YL-10),枯草芽孢杆菌(B.subtilis YL-09),地衣芽孢杆菌(B.licheniformis ge6-1)和巨大芽孢杆菌(B.megaterium H1)分别进行发酵培养,测定其单株芽孢杆菌发酵液和多株混合芽孢杆菌发酵液中的消化酶活性以及胞外酶产生情况,从中筛选酶活性高和稳定性好的实验组分别进行凡纳滨对虾(Litopenaeus vannamei)养殖实验。研究结果显示:单株芽孢杆菌YL-10具有最高的蛋白酶和淀粉酶活性,分别为335.0和3.0U/mL;ge6-1具有最高的脂肪酶活性,为5.0U/mL。混合芽孢杆菌Tm1,Tm2,Tm3和Tm5发酵液中蛋白酶活性和淀粉酶活性均高于单株芽孢杆菌ge6-1和H1。通过API zym系统测定芽孢杆菌胞外酶的产生情况,发现多株混合芽孢杆菌发酵液中胞外酶的种类多于单株芽孢杆菌。凡纳滨对虾养殖实验结果显示:YL-10组(解淀粉芽孢杆菌)与Tm3组(解淀粉芽孢杆菌+枯草芽孢杆菌+地衣芽孢杆菌)对虾的体重增加、特定生长率和存活率较对照组显著提高(P<0.05)。芽孢杆菌能够提高对虾肝胰腺中的消化酶活性:YL-10组淀粉酶活性显著高于对照组,Tm3组蛋白酶和脂肪酶活性显著高于对照组。API zym系统测定结果显示:YL-10组和Tm3组对虾肝胰腺中分别有10种和12种胞外酶的酶活性提高,但各处理组间胞外酶活性差异不显著(P>0.05)。
Abstract：This study profiled the enzyme activities of single and mixed cultures of Bacillus subtilis YL-09,B.licheniformis ge6-1,B.megaterium H1 and B.amyloliquefaciens YL-10 using API zym system and digestive enzyme assay,and two Bacillus cultures with higher enzyme activities and better stability were selected for feeding assay.From the result of digestive enzyme assay,B.amyloliquefaciens YL-10 showed the highest protease and amylase activities(315.0 and 3.0 U·mL-1)and B.licheniformis ge6-1 was the best lipase producer,with lipase activity of 5.0 U·mL-1.Higher production of protease and lipase was detected in mixed Bacillus cultures Tm1,Tm2,Tm3 and Tm5 compared with single Bacillus H1 and ge6-1.Different enzyme patterns of the selected Bacillus cultures(single and mixed Bacillus cultures)were detected using API zym system.And the enzyme patterns resulting from API zym also indicated that mixed Bacillus cultures showed the more diversified enzyme production.Shrimps fed YL-10(B.amyloliquefaciens YL-10)and mixed Bacillus culture Tm3(B.amyloliquefaciens YL-10 + B.subtilis YL-09 + B.licheniformis ge6-1)exhibited significantly(P < 0.05)higher weight gain(WG),specific growth rate(SGR)and survival rate(SR).The digestive enzyme analyses showed diet containing Tm3(B.amyloliquefaciens YL-10 + B.subtilis YL-09 + B.licheniformis ge6-1)increased protease and lipase activities in hepatopancreas while diet with YL-10(B.amyloliquefaciens YL-10)increased amylase activity at the end of feeding(P < 0.05).Constitutive enzyme activities resulted from API zym system showed the 10 kinds of extracellular enzymes were recorded in group YL-10 while 12 kinds were recorded in Tm3.
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基本信息:
DOI：10.16441/j.cnki.hdxb.20160404
中图分类号:S968.22
引用信息:
[1]游龙,韩茵,张凯,等.芽孢杆菌产胞外酶的活性分析及其对凡纳滨对虾的作用[J],2018,48(03):80-87.DOI:10.16441/j.cnki.hdxb.20160404.
基金信息:
国家自然科学基金项目(31072241)资助~~