纤维素酶高产菌株选育及发酵条件的优化
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摘要
本研究首先对酶活较高的菌株3.4261进行亚硝酸、紫外线和硫酸二乙酯复合诱变,然后通过羧甲基纤维素钠培养基、刚果红纤维素培养基进行初筛和液态发酵复筛,得到2株酶活较高的菌株,命名为E20-2和E40-4,对相对活性更高的E20-2进行发酵条件优化,其最适条件为麸皮与麦秸杆比为3:7,固液比为1/4～1/3,发酵时间5天,初始pH6.0,Tween-80浓度0.1%,培养温度28℃,其CMCase和FPA最高分别可达654U/g和69U/g,分别较出发菌株提高230%和72%。
另外,从甘肃省玛曲县采样12份,用滤纸为碳源富集然后以羧甲基纤维素钠培养基和刚果红纤维素培养基进行初筛,筛选出水解圈直径和菌落直径比较大的产纤维素酶菌株60余株,然后经过液态发酵复筛,通过测CMCase和FPA得到较高的野生菌株有5株,其中酶活最高的命名为G7,其最适产酶条件:CMCNa最适浓度0.4%,(NH_4)_2SO_40.15%,最适温度28℃,pH6.0,发酵时间4天,其FPA和CMCase分别达到1.767U/mL和7.5243U/mL。
最后,比较E20-2单菌种发酵及混合发酵产酶差异,结果显示E20-2混菌固体发酵试验比单菌种发酵产酶明显要低。
Cellulase-producing 3.4261 was chosen as the original strain and treated with UV and DES, two high cellulase-producing mutants,named E20-2 and E40-4 were obtained by screening with transparent zone method and shaking culture method.Preliminary analysis of optimal fermentation conditions of E20-2 showed that the process required ratio of wheat bran and wheat straw was 3 to 7,ratio of solid and liquid was 1/4 to 1/3,Tween80 concentration was 0.1%,temperature 28℃,pH 6.0,cultured for 5 days.The CMCase and FPA of the mutant strain reached 654U/g and 69U/g respectively,which were 230%and 72%higher than the orginal strain.
More than sixty strains were obtained by screening with transparent zone method from the Maqu county of China where the climate was special.Then through shaking culture method,five high cellulase-producing stains were obtained,the highest cellulase-producing strain named G7.Preliminary analysis of submerged fermentation conditions showed that the optimal concentration of CMCNa was 0.4%,(NH_4)_2SO_4 0.15%,pH 6.0,and a temperature range between 28℃and 30℃,cultured for 4 days.The CMCase and FPA of G7 reached 1.767U/mL and 7.524U/mL.
Results indicated that E20-2's isolate fermentation was more effective than mixed fermentation.
另外,从甘肃省玛曲县采样12份,用滤纸为碳源富集然后以羧甲基纤维素钠培养基和刚果红纤维素培养基进行初筛,筛选出水解圈直径和菌落直径比较大的产纤维素酶菌株60余株,然后经过液态发酵复筛,通过测CMCase和FPA得到较高的野生菌株有5株,其中酶活最高的命名为G7,其最适产酶条件:CMCNa最适浓度0.4%,(NH_4)_2SO_40.15%,最适温度28℃,pH6.0,发酵时间4天,其FPA和CMCase分别达到1.767U/mL和7.5243U/mL。
最后,比较E20-2单菌种发酵及混合发酵产酶差异,结果显示E20-2混菌固体发酵试验比单菌种发酵产酶明显要低。
Cellulase-producing 3.4261 was chosen as the original strain and treated with UV and DES, two high cellulase-producing mutants,named E20-2 and E40-4 were obtained by screening with transparent zone method and shaking culture method.Preliminary analysis of optimal fermentation conditions of E20-2 showed that the process required ratio of wheat bran and wheat straw was 3 to 7,ratio of solid and liquid was 1/4 to 1/3,Tween80 concentration was 0.1%,temperature 28℃,pH 6.0,cultured for 5 days.The CMCase and FPA of the mutant strain reached 654U/g and 69U/g respectively,which were 230%and 72%higher than the orginal strain.
More than sixty strains were obtained by screening with transparent zone method from the Maqu county of China where the climate was special.Then through shaking culture method,five high cellulase-producing stains were obtained,the highest cellulase-producing strain named G7.Preliminary analysis of submerged fermentation conditions showed that the optimal concentration of CMCNa was 0.4%,(NH_4)_2SO_4 0.15%,pH 6.0,and a temperature range between 28℃and 30℃,cultured for 4 days.The CMCase and FPA of G7 reached 1.767U/mL and 7.524U/mL.
Results indicated that E20-2's isolate fermentation was more effective than mixed fermentation.
引文
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