土壤中产高活性淀粉酶细菌的分离与纯化
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摘要
[目的]从土壤中分离纯化产高活性淀粉酶细菌,并研究其产酶条件。[方法]利用淀粉水解圈平板筛选法,从都匀市23份不同地点的土样中分离并筛选到一株产胞外淀粉酶的菌株Z_3,其水解圈/菌落直径比达5.50;通过形态学、染色等方法,结合16S r DNA序列的比对鉴定该菌株为蜡状芽孢杆菌(Bacillus cereus)。并对Z_3菌株产酶条件进行了研究。[结果]Z3菌株在1.0%淀粉的碳源、1.0%酵母膏+1.0%牛肉膏的氮源、0.2%的Ca Cl_2·2H_2O、30 m L装液量于250 m L摇瓶中的溶氧量,p H为7.0,培养温度为37℃的条件下,产酶能力最强。此外,Z3菌株所产淀粉酶的酶学分析显示,其最适酶反应p H为8.0,最适反应温度为39℃。[结论]该菌株是产淀粉酶较好的材料,具有一定的应用前景。
[Objective]Bacteria producing highly active amylase in soil was isolated and purificated,and enzyme production conditions were studied.[Method]Using starch hydrolysis circle model,a amylase-producing strain Z3 was screened from 23 soil samples in Duyun. The hydrolysis circle/colony diameter ratio were 5.50; Z3 was identified as Bacillus cereus strain by the morphological observation,staining and 16 S r DNA sequences alignment analysis. [Result]The optimum carbon source,nitrogen source,concentration of CaCl2·2 H2 O,dissolved oxygen,p H and temperature was 1% starch,1% yeast extract+1% beef extract,0.2% CaCl2·2 H2 O,adding 30 m L medium into 250 m L flask,p H 7.0 and 37 ℃ respectively. Besides,the enzymatic analysis of Z3 amylase showed that its optimum p H and temperature was 8. 0 and 39 ℃ respectively. [Conclusion]This study showed that Z3 strain was a good amylase-producing material for the further industrial utilization.
[Objective]Bacteria producing highly active amylase in soil was isolated and purificated,and enzyme production conditions were studied.[Method]Using starch hydrolysis circle model,a amylase-producing strain Z3 was screened from 23 soil samples in Duyun. The hydrolysis circle/colony diameter ratio were 5.50; Z3 was identified as Bacillus cereus strain by the morphological observation,staining and 16 S r DNA sequences alignment analysis. [Result]The optimum carbon source,nitrogen source,concentration of CaCl2·2 H2 O,dissolved oxygen,p H and temperature was 1% starch,1% yeast extract+1% beef extract,0.2% CaCl2·2 H2 O,adding 30 m L medium into 250 m L flask,p H 7.0 and 37 ℃ respectively. Besides,the enzymatic analysis of Z3 amylase showed that its optimum p H and temperature was 8. 0 and 39 ℃ respectively. [Conclusion]This study showed that Z3 strain was a good amylase-producing material for the further industrial utilization.
引文
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[2]易征璇,赵彤,师旋,等.高温淀粉酶高产菌株筛选及产酶条件优化[J].现代农业科技,2014(7):240-243.
[3]陈约慧,陈颖聪,刘冬冬,等.一株高产淀粉酶的解淀粉芽胞杆菌的分离鉴定及其产酶条件优化[J].中国微生态学杂志,2011,23(6):505-515.
[4]王鑫昕,耿霄,吴子龙,等.丛台酒大曲中高产淀粉酶细菌的分离和鉴定[J].酿酒科技,2017(1):30-32.
[5]吕晓龙.海参养殖水体病原微生物生物治理技术研究[D].大连:大连海事大学,2012.
[6]代先祝.低营养物浓度培养基分离淀粉酶分泌细菌及其特性的初步研究[J].酿酒科技,2011(11):26-29.
[7]陈源源,牛丹丹,王正祥.一种快速鉴定细菌的方法[J].食品与发酵工业,2007,33(5):29-31.
[8]雷晓燕.土壤中淀粉酶产生菌的筛选及产酶条件优化[J].沈阳化工大学学报,2010,24(3):203-208.
[9]孙佑赫,周开艳,熊智.一株产α-淀粉酶细菌的分离、鉴定及酶学性质初步研究[J].华北农学报,2012,27(S1):250-253.